Biochar-nanoparticle combinations enhance the biogeochemical recovery of a post-mining soil.

Here we addressed the capacity of distinct amendments to reduce arsenic (As), copper (Cu), selenium (Se) and zinc (Zn) associated risks and improve the biogeochemical functions of post-mining soil. To this, we examined nanoparticles (NPs) and/or biochar effects, combined with phytostabilization using Lolium perenne L. Soil samples were taken in a former metal mine surroundings. Ryegrass seeds were sown in pots containing different combinations of NPs (zero-valent iron (nZVI) or hydroxyapatite (nH)) (0 and 2 %), and biochar (0, 3 and 5 %). Plants were grown for 45 days and the plant yield and element accumulation were evaluated, also soil properties (element distribution within the soil fractions, fertility, and enzymatic activities associated with microbiota functionality and nutrient cycling) were determined. Results showed biochar-treated soil had a higher pH, and much higher organic carbon (C) content than control soil and NP-treated soils, and it revealed increased labile C, total N, and available P concentrations. Soil treatment with NP-biochar combinations increased exchangeable non-acid cation concentrations and reduced exchangeable Na%, improved soil fertility, reduced sodicity risk, and increased ryegrass biomass. Enzymatic activities, particularly dehydrogenase and glucosidase, increased upon the addition of biochar, and this effect was fostered by NPs. Most treatments led to a significant reduction of metal(loid)s contents in biomass, mitigating contamination risks. The two different NPs had similar effects in many parameters, nH outperformed nZVI in terms of increased nutrients, C content, and enzymatic activities. On the basis of our results, combined biochar-NP amendments use, specially nH, emerges as a potential post-mining soil restoration strategy.

Phytoremediation potential depends on the degree of soil pollution: a case study in an urban brownfield.

Phytoremediation is a cost-effective nature-based solution for brownfield reclamation. The choice of phytoextraction or phytostabilization strategies is highly relevant when planning full-scale treatments. A suitable approach to identify such species involves the evaluation of plants that grow spontaneously on the contaminated sites. Here, we sought to determine the phytoremediation potential of three spontaneous plant species, namely the trees Acer pseudoplatanus L (A. pseudoplatanus) and Betula celtiberica Rothm. & Vasc (B. celtiberica), and the shrub Buddleja davidii Franch (B. davidii), for the recovery of an urban brownfield. To determine the response of the species to the degree of contamination, we conducted soil and vegetation sampling inside and outside the site. The concentrations of As, Cu, and Zn in soil and plant samples were measured, and then various indexes related to phytoremediation were calculated. The translocation factor and transfer coefficient indicated that vegetation outside the brownfield had phytoextraction capacity while the same plants inside the brownfield revealed phytostabilization properties. Given our results, we propose that the selected species are suitable for phytostabilization strategies in areas with high concentrations of contaminants, whereas they could be used for phytoextraction only in soils with low or moderate levels of pollution.

The microbiome of a brownfield highly polluted with mercury and arsenic.

Abandoned brownfields represent a challenge for their recovery. To apply sustainable remediation technologies, such as bioremediation or phytoremediation, indigenous microorganisms are essential agents since they are adapted to the ecology of the soil. Better understanding of microbial communities inhabiting those soils, identification of microorganisms that drive detoxification process and recognising their needs and interactions will significantly improve the outcome of the remediation. With this in mind we have carried out a detailed metagenomic analysis to explore the taxonomic and functional diversity of the prokaryotic and eukaryotic microbial communities in soils, several mineralogically distinct types of pyrometallurgic waste, and groundwater sediments of a former mercury mining and metallurgy site which harbour very high levels of arsenic and mercury pollution. Prokaryotic and eukaryotic communities were identified, which turned out to be more diverse in the surrounding contaminated soils compared to the pyrometallurgic waste. The highest diversity loss was observed in two environments most contaminated with mercury and arsenic (stupp, a solid mercury condenser residue and arsenic-rich soot from arsenic condensers). Interestingly, microbial communities in the stupp were dominated by an overwhelming majority of archaea of the phylum Crenarchaeota, while Ascomycota and Basidiomycota fungi comprised the fungal communities of both stump and soot, results that show the impressive ability of these previously unreported microorganisms to colonize these extreme brownfield environments. Functional predictions for mercury and arsenic resistance/detoxification genes show their increase in environments with higher levels of pollution. Our work establishes the bases to design sustainable remediation methods and, equally important, to study in depth the genetic and functional mechanisms that enable the subsistence of microbial populations in these extremely selective environments.

Pyrolysis temperature influences the capacity of biochar to immobilize copper and arsenic in mining soil remediation.

Biochar is a promising material used for multiple remediation approaches, mainly in polluted soils. Its properties can differ depending on feedstock and pyrolysis temperature. In this context, we tested the capacity of three biochar products made from corncob, pyrolyzed at different temperatures (350, 500, and 650 °C), to remediate a mining soil affected by high levels of Cu and As. We performed an exhaustive characterization of the biochar. We found that biochar showed a higher surface area with increasing pyrolysis temperature, whereas high molecular weight PAHs were detected in biochar produced at the maximum temperature, thus indicating potential ecotoxicological risks. After the application of biochar to the soil, Cu was partially immobilized, especially when using that obtained at 500 °C. This effect is attributed to the structure of this material and an increase in soil pH and organic matter content. Conversely, As was increased in the soluble fraction for all three types of biochar but in a proportion that lacks relevance. On the whole, given its lower PAH content, higher Cu immobilization ratio, and an almost negligible increase in As availability, biochar obtained at 500 °C outperformed the other two products with respect to soil recovery. Of note, data on Cu and As availability were doubled-checked using two extraction methodologies. We propose that this operational approach for determining the most suitable pyrolysis temperature will find application in other soil remediation actions.

Effective bioremediation of soil from the Burgan oil field (Kuwait) using compost: A comprehensive hydrocarbon and DNA fingerprinting study.

An innovative combination of metagenomic profiling of microbial communities and GC-MS & Pyrolysis-GC-MS fingerprinting methods were used to assess the biodegradation of contaminated soil from the Burgan oil field in Kuwait. The soil was treated with (sludge) compost in microcosms to evaluate the feasibility of this material for bioremediation purposes. The most favourable trial showed a > 80% decrease in TPH, thereby indicating strong potential for full-scale application using a cost-effective technology and thus in line with the principles of the circular economy. The microbial study showed that compost addition enhanced the organic matter and nutrient content of the soil. However, the microorganisms in the compost did not seem to play a relevant role in bioremediation, meaning that compost amendments serve as a biostimulation rather than a bioaugmentation approach. The chemical study of the distinct oil fractions revealed rapidly biodegraded compounds (alkanes, alkyl-aromatics, etc.) and others that were much more refractory (hopanes, benzohopanes, etc.). Of note, although heavy fractions are usually considered recalcitrant to biodegradation, we observed incipient degradation of the asphaltene fraction by means of double-shot thermodesorption and pyrolysis. Finally, chemical fingerprinting also revealed that the treated soil contained some of the compounds found in the compost, such as coprostanol, cholesterol, and plant sterols. This observation would support the use of these compounds as proxies to monitor the effects of compost and to adjust dosages in real-scale bioremediation treatments.

Evaluation of biostimulation, bioaugmentation, and organic amendments application on the bioremediation of recalcitrant hydrocarbons of soil.

In the present work, the operational conditions for improving the degradation rates of Total Petroleum Hydrocarbons (TPHs) in contaminated soil from a machinery park were optimized at a microcosms scale along a 90-days incubation period. In this study, bioremediation strategies and an organic amendment have been tested to verify the remediation of soil contaminated with different hydrocarbons, mineral oils, and heavy metals. Specifically, designed biostimulation and bioaugmentation strategies were compared with and without adding vermicompost. The polluted soil harboring multiple contaminants, partially attenuated for years, was used. The initial profile showed enrichment in heavy linear alkanes, suggesting a previous moderate weathering. The application of vermicompost increased five and two times the amounts of available phosphorus (P) and exchangeable potassium (K), respectively, as a direct consequence of the organic amendment addition. The microbial activity increased due to soil acidification, which influenced the solubility of P and other micronutrients. It also impacted the predominance and variability of the different microbial groups and the incubation, as reflected by phospholipid fatty acid (PLFA) results. An increase in the alkaline phosphatases and proteases linked to bacterial growth was displayed. This stimulation of microbial metabolism correlated with the degradation rates since TPHs degradation' efficiency after vermicompost addition reached 32.5% and 34.4% of the initial hydrocarbon levels for biostimulation and bioaugmentation, respectively. Although Polycyclic Aromatic Hydrocarbons (PAHs) were less abundant in this soil, results also decreased, especially for the most abundant, the phenanthrene. Despite improving the degradation rates, results revealed that recalcitrant and hydrophobic petroleum compounds remained unchanged, indicating that mobility, linked to bioavailability, probably represents the limiting step for further soil recovery.

Contrasting mobility of arsenic and copper in a mining soil: A comparative column leaching and pot testing approach.

The remediation of legacy metal(loid) contaminated soils in-situ relies on the addition of [organic] amendments to reduce the mobility and bioavailability of metal(loid)s, improve soil geochemical parameters and restore vegetation growth. Two vermicomposts of food and animal manure waste origin (V1 and V2) were amended to an arsenic (As) and copper (Cu) contaminated mine soil (≤1500 mg kg-1). Leaching columns and pot experiments evaluated copper and arsenic in soil pore waters, as well as pH, dissolved organic carbon (DOC) and phosphate (PO43-) concentrations. The uptake of As and Cu to ryegrass was also measured via the pot experiment, whilst recovered biochars from the column leaching test were measured for metal sorption at the termination of leaching. Vermicompost amendment to soil facilitated ryegrass growth which was entirely absent from the untreated soil in the pot test. All amendment combinations raised pore water pH by ∼4 units. Copper concentrations in pore waters from columns and pots showed steep reductions (∼1 mg L-1), as a result of V1 & V2 compared to untreated soil (∼500 mg L-1). Combined with an increase in DOC and PO43-, As was mobilised an order of magnitude by V1. Biochar furthest reduced Cu in pore waters from the columns to <0.1 mg L-1, as a result of surface sorption. The results of this study indicate that biochar can restrict the mobility of Cu from a contaminated mine soil after other amendment interventions have been used to promote revegetation. However, the case of As, biochar cannot counter the profound impact of vermicompost on arsenic mobility.

Interplay between arsenic and selenium biomineralization in Shewanella sp. O23S.

Bacteria play crucial roles in the biogeochemical cycle of arsenic (As) and selenium (Se) as these elements are metabolized via detoxification, energy generation (anaerobic respiration) and biosynthesis (e.g. selenocysteine) strategies. To date, arsenic and selenium biomineralization in bacteria were studied separately. In this study, the anaerobic metabolism of As and Se in Shewanella sp. O23S was investigated separately and mixed, with an emphasis put on the biomineralization products of this process. Multiple analytical techniques including ICP-MS, TEM-EDS, XRD, Micro-Raman, spectrophotometry and surface charge (zeta potential) were employed. Shewanella sp. O23S is capable of reducing selenate (SeO42-) and selenite (SeO32-) to red Se(-S)0, and arsenate (AsO43-) to arsenite (AsO33-). The release of H2S from cysteine led to the precipitation of AsS minerals: nanorod AsS and granular As2S3. When As and Se oxyanions were mixed, both As-S and Se(-S)0 biominerals were synthesized. All biominerals were extracellular, amorphous and presented a negative surface charge (-24 to -38 mV). Kinetic analysis indicated the following reduction yields: SeO32- (90%), AsO43- (60%), and SeO42- (<10%). The mix of SeO32- with AsO43- led to a decrease in As removal to 30%, while Se reduction yield was unaffected (88%). Interestingly, SeO42- incubated with AsO43- boosted the Se removal (71%). The exclusive extracellular formation of As and Se biominerals might indicate an extracellular respiratory process characteristic of various Shewanella species and strains. This is the first study documenting a complex interplay between As and Se oxyanions: selenite decreased arsenate reduction, whereas arsenate stimulated selenate reduction. Further investigation needs to clarify whether Shewanella sp. O23S employs multi-substrate respiratory enzymes or separate, high affinity enzymes for As and Se oxyanion respiration.

Effects of in situ Remediation With Nanoscale Zero Valence Iron on the Physicochemical Conditions and Bacterial Communities of Groundwater Contaminated With Arsenic.

Nanoscale Zero-Valent Iron (nZVI) is a cost-effective nanomaterial that is widely used to remove a broad range of metal(loid)s and organic contaminants from soil and groundwater. In some cases, this material alters the taxonomic and functional composition of the bacterial communities present in these matrices; however, there is no conclusive data that can be generalized to all scenarios. Here we studied the effect of nZVI application in situ on groundwater from the site of an abandoned fertilizer factory in Asturias, Spain, mainly polluted with arsenic (As). The geochemical characteristics of the water correspond to a microaerophilic and oligotrophic environment. Physico-chemical and microbiological (cultured and total bacterial diversity) parameters were monitored before and after nZVI application over six months. nZVI treatment led to a marked increase in Fe(II) concentration and a notable fall in the oxidation-reduction potential during the first month of treatment. A substantial decrease in the concentration of As during the first days of treatment was observed, although strong fluctuations were subsequently detected in most of the wells throughout the six-month experiment. The possible toxic effects of nZVI on groundwater bacteria could not be clearly determined from direct observation of those bacteria after staining with viability dyes. The number of cultured bacteria increased during the first two weeks of the treatment, although this was followed by a continuous decrease for the following two weeks, reaching levels moderately below the initial number at the end of sampling, and by changes in their taxonomic composition. Most bacteria were tolerant to high As(V) concentrations and showed the presence of diverse As resistance genes. A more complete study of the structure and diversity of the bacterial community in the groundwater using automated ribosomal intergenic spacer analysis (ARISA) and sequencing of the 16S rRNA amplicons by Illumina confirmed significant alterations in its composition, with a reduction in richness and diversity (the latter evidenced by Illumina data) after treatment with nZVI. The anaerobic conditions stimulated by treatment favored the development of sulfate-reducing bacteria, thereby opening up the possibility to achieve more efficient removal of As.

Application of biochar, compost and ZVI nanoparticles for the remediation of As, Cu, Pb and Zn polluted soil.

Here we tested the capacity of zero valent iron nanoparticles (nZVI) combined with two organic amendments, namely, compost and biochar, to immobilize metal(oid)s such as As, Cu, Pb, and Zn. In addition, the effects of the amendments on the development of Brassica juncea L., a plant widely used for phytoremediation purposes, were also examined. To perform the experiments, pots containing polluted soil were treated with nZVI, compost-biochar, or a blend of compost-biochar-nZVI. Metal(oid)s availability and soil properties were evaluated after 15 and 75 days, and the height and weight of the plants were measured to determine development. The compost-biochar amendment showed excellent capacity to immobilize metals, but As availability was considerably increased. However, the addition of nZVI to the mixture corrected this effect considerably. In addition, soil treatment with nZVI alone led to a slight increase in Cu availability, which was not observed for the mixture with organic amendments. With respect to soil properties, the CEC and pH were enhanced by the compost-biochar amendment, thereby favoring plant growth. Nevertheless, the nanoparticles reduced the concentration of available P, which impaired plant growth to a certain extent. In conclusion, Fe-based nanoparticles combined with organic amendments emerge as powerful approaches to remediate soils contaminated by metals and metalloids.

A multi-faceted, environmental forensic characterization of a paradigmatic brownfield polluted by hazardous waste containing Hg, As, PAHs and dioxins.

Hg and As mining-metallurgy plants have severely impacted environmental compartments. La Soterraña site (northern Spain) has been previously studied in this context. However, here we used a novel multi-purpose forensic approach to examine accumulations not only of mining-metallurgical waste (volumes above 80,000 t) but also C&D waste as a repository of pollutants (above 10% of As leached in standard tests) at this site. High Hg and As content in very fine grain-size fractions (up to 100,000 mg/kg of As in metallurgy waste below 10 µm) was significant, as was the predominance of As (III) in metallurgy waste. In addition, GC-MS techniques revealed the predominance of PAHs (secondarily Oxy-PAHs and PCBs), which showed a pyrogenic fingerprint, as determined by molecular ratios. Moreover, toxic organometallics (Hg-aromatics) were detected and metallurgy waste was identified as a source of dioxins and furanes (TEQ close to 30). On the basis of our observations, La Soterraña emerges as one of the most polluted sites in Europe and therefore requires urgent remediation. Our key findings indicate that C&D waste should be considered hazardous. Metallurgy waste, in turn, raises maximum concern given the simultaneous presence of toxic inorganic and organic contaminants.

Nanoremediation of As and metals polluted soils by means of graphene oxide nanoparticles.

The capacity of graphene oxide nanoparticles (nGOx) to reduce or increase As and metals availability in polluted soils was compared with that of zero valent iron nanoparticles (nZVI). The nanomaterials used in this study were characterized by X-ray techniques, CHNS-O analysis, dynamic light scattering, and microscopy procedures such as atomic force microscopy. To assess the capacity of these materials to immobilize pollutants, field samples of two soils were treated with nZVI and nGOx at a range of doses (0.2%, 1% and 5%). Availability tests were then performed. nGOx effectively immobilized Cu, Pb and Cd, but mobilized As and P (even at low doses), in the latter case irrespective of the simultaneous presence of high concentrations of metals. In turn, nZVI promoted notable immobilization results for As and Pb, a poorer result for Cd, and an increased availability for Cu. Soil pH and EC have been slightly affected by nGOx. On the whole, nGOx emerges as a promising option for mobilization/immobilization strategies for soil nanoremediation when combined with other techniques such as phytoremediation.

Arsenic release from pyrite ash waste over an active hydrogeological system and its effects on water quality.

Arsenic is a potentially toxic element of concern for environmental compartments, and it is a frequent pollutant in many abandoned industrial sites. In this study, geochemical and hydrogeological tools were used to determine the long-term effects of As-rich pyrite ash disposal (83,000 m3 as estimated by geostatistical tools) in a brownfield located over a quaternary alluvial aquifer. Throughout the site, soil pollution and water table oscillation led to leachates in the form of both run-off and infiltration waters, thereby reducing (ground)water quality (e.g. pH, electrical conductivity) and, in particular, increasing the concentration of arsenic (average approx. 4000 µg/l for one hydrological year). By means of laboratory and in situ measurements, the main mechanisms through which the sulphide remaining in the pyrite ash leaches were identified. In addition, to evaluate the effects of the polluted groundwater on the nearby main river, a mathematical approach using the Domenico analytical groundwater transport model revealed potential concentrations of 49 µg/l of arsenic in the junction between the study aquifer and the river, equivalent to an annual quantity of 49 kg of this element.

Diagnostic ratios for the rapid evaluation of natural attenuation of heavy fuel oil pollution along shores.

Marine oil spills are typically followed by complex clean-up and monitoring operations of the shorelines affected. In this regard, determination of the changes in the chemical composition of the spilled product is crucial for evaluation purposes. The "GC-MS fingerprint" approach requires the identification of several key parameters in order to differentiate between weathering processes. In this context, we performed a 900-day study (eight sampling campaigns) of natural attenuation on a rocky shore affected by a heavy fuel oil spill. The residues coating the rocks were studied by extraction, separation and evaluation of SARA fractions, followed by GC-MS analysis for saturates and aromatics. The results identified a group of diagnostic ratios with irregular capabilities to differentiate between volatilization, photodegradation, and biodegradation (using n-alkanes, isoprenoids and PAHs), while a second group of primarily stable ratios (using dibenzothiphenes, tricyclics and hopanes) was also obtained. Notably, this is the first time that some of these ratios have been used for marine spill monitoring purposes. The ratios were applied not only to evaluate weathering at the study site, but also to obtain a comparison with the degradation of floating oil slicks-a demonstration that weathering of the latter was quicker than that of oil on the shore rocks. Additionally, Pyrolysis-GC-MS analysis of the asphaltene fraction of residues coating the rocks revealed moderate changes in the composition of this initially recalcitrant fraction. These changes were consistent with those found in the proportion of resins and asphaltene fractions over time.

Use of Endophytic and Rhizosphere Bacteria To Improve Phytoremediation of Arsenic-Contaminated Industrial Soils by Autochthonous Betula celtiberica.

The aim of this study was to investigate the potential of indigenous arsenic-tolerant bacteria to enhance arsenic phytoremediation by the autochthonous pseudometallophyte Betula celtiberica The first goal was to perform an initial analysis of the entire rhizosphere and endophytic bacterial communities of the above-named accumulator plant, including the cultivable bacterial species. B. celtiberica's microbiome was dominated by taxa related to Flavobacteriales, Burkholderiales, and Pseudomonadales, especially the Pseudomonas and Flavobacterium genera. A total of 54 cultivable rhizobacteria and 41 root endophytes, mainly affiliated with the phyla Proteobacteria, Bacteroidetes, Firmicutes, and Actinobacteria, were isolated and characterized with respect to several potentially useful features for metal plant accumulation, such as the ability to promote plant growth, metal chelation, and/or mitigation of heavy-metal stress. Seven bacterial isolates were further selected and tested for in vitro accumulation of arsenic in plants; four of them were finally assayed in field-scale bioaugmentation experiments. The exposure to arsenic in vitro caused an increase in the total nonprotein thiol compound content in roots, suggesting a detoxification mechanism through phytochelatin complexation. In the contaminated field, the siderophore and indole-3-acetic acid producers of the endophytic bacterial consortium enhanced arsenic accumulation in the leaves and roots of Betula celtiberica, whereas the rhizosphere isolate Ensifer adhaerens strain 91R mainly promoted plant growth. Field experimentation showed that additional factors, such as soil arsenic content and pH, influenced arsenic uptake in the plant, attesting to the relevance of field conditions in the success of phytoextraction strategies.IMPORTANCE Microorganisms and plants have developed several ways of dealing with arsenic, allowing them to resist and metabolize this metalloid. These properties form the basis of phytoremediation treatments and the understanding that the interactions of plants with soil bacteria are crucial for the optimization of arsenic uptake. To address this in our work, we initially performed a microbiome analysis of the autochthonous Betula celtiberica plants growing in arsenic-contaminated soils, including endosphere and rhizosphere bacterial communities. We then proceeded to isolate and characterize the cultivable bacteria that were potentially better suited to enhance phytoextraction efficiency. Eventually, we went to the field application stage. Our results corroborated the idea that recovery of pseudometallophyte-associated bacteria adapted to a large historically contaminated site and their use in bioaugmentation technologies are affordable experimental approaches and potentially very useful for implementing effective phytoremediation strategies with plants and their indigenous bacteria.

Human health risk assessment in restoring safe and productive use of abandoned contaminated sites.

In Europe soil contamination has been recognized as a serious problem. The needs to remediate contaminated sites are not questionable, although the remediation actions are often hindered by their very high financial costs. On the other hand, the abandoned contaminated sites may have the potential for redevelopment and creating conditions appropriate for their productive reuse bringing social, economic and environmental benefits. The main concern associated with the contaminated sites is their potential adverse health impact. Therefore, in the process of contaminated site redevelopment the risk assessment and the subsequent risk management decisions will play a crucial role. The main objective of this study was to illustrate the role of the human health risk assessment (HRA) in supporting site remediation and reuse decisions. To exemplify the significance of the HRA process in this field the Nitrastur site, located in Asturias, Spain was used. Risks resulting from soil contamination with arsenic (As), cadmium (Cd), chromium (Cr), copper (Cu), nickel (Ni), mercury (Hg), zinc (Zn) and lead (Pb) were assessed under three potential future land use patterns: industrial, residential and recreational. The results of the study indicated that soil at the Nitrastur site might pose non-cancer and cancer risks to potential future receptors - industrial workers, residents and recreational users. Arsenic and lead are the main substances responsible for the health risk and the primary drivers of remedial decisions at the site. The highest total cancer risks were observed under the residential scenario, followed in descending order by the recreational and industrial ones. The remedial maps illustrate in which areas remediation activities are required, depending on a given land use pattern. The obtained results may be used to develop, analyse, compare and select the remedial options within the intended land use pattern. They may also be used to support the decisions concerning the reuse of the site or to create different redevelopment plans for different parts of the site. The study may help improve understanding and interaction between risk assessors and risk managers which are necessary to make proper, fully informed decisions enhancing public trust and credibility.

Multivariate study of trace element distribution in the geological record of Roñanzas Peat Bog (Asturias, N. Spain). Paleoenvironmental evolution and human activities over the last 8000 calyr BP.

Trace element concentrations in the Roñanzas peat bog record reveal a contribution of natural processes but the influence of anthropogenic factors predominates in the last two millenniums, particularly aerosol deposition linked to mining and industrial activities in northern Spain. We observed that the Roñanzas record can be considered a preserved environment, suitable to search for local (<50 km), regional (50-150 km) and/or long-distance human activity fingerprinting, specifically that related to the deposition of heavy metals such as Pb, Zn and Hg. We also carried out a multivariate statistical study in order to clarify the geochemical behavior of trace and major elements. Our study design represents a novel approach to assign natural vs. human contributions in peatlands. Therefore, synergies obtained by the simultaneous study of multivariate statistics and enrichment factors allow robust conclusions about paleoenvironmental evolution and human activities. Anthropogenic influence has also been reported in similar records in other parts of Europe, thereby suggesting large-scale sources for atmospheric pollution. However, here we revealed remarkable particularities, such as the association of Cd, Zn and Pb, mainly linked to regional and local factors (mining and more recently the metallurgical industry), whereas we propose that the occurrence of Hg is associated with a combination of regional factors and global atmospheric pollution.

Metaproteogenomic insights beyond bacterial response to naphthalene exposure and bio-stimulation.

Microbial metabolism in aromatic-contaminated environments has important ecological implications, and obtaining a complete understanding of this process remains a relevant goal. To understand the roles of biodiversity and aromatic-mediated genetic and metabolic rearrangements, we conducted 'OMIC' investigations in an anthropogenically influenced and polyaromatic hydrocarbon (PAH)-contaminated soil with (Nbs) or without (N) bio-stimulation with calcium ammonia nitrate, NH(4)NO(3) and KH(2)PO(4) and the commercial surfactant Iveysol, plus two naphthalene-enriched communities derived from both soils (CN2 and CN1, respectively). Using a metagenomic approach, a total of 52, 53, 14 and 12 distinct species (according to operational phylogenetic units (OPU) in our work equivalent to taxonomic species) were identified in the N, Nbs, CN1 and CN2 communities, respectively. Approximately 10 out of 95 distinct species and 238 out of 3293 clusters of orthologous groups (COGs) protein families identified were clearly stimulated under the assayed conditions, whereas only two species and 1465 COGs conformed to the common set in all of the mesocosms. Results indicated distinct biodegradation capabilities for the utilisation of potential growth-supporting aromatics, which results in bio-stimulated communities being extremely fit to naphthalene utilisation and non-stimulated communities exhibiting a greater metabolic window than previously predicted. On the basis of comparing protein expression profiles and metagenome data sets, inter-alia interactions among members were hypothesised. The utilisation of curated databases is discussed and used for first time to reconstruct 'presumptive' degradation networks for complex microbial communities.

Biodegradation of oil tank bottom sludge using microbial consortia.

We present a rationale for the selection of a microbial consortia specifically adapted to degrade toxic components of oil refinery tank bottom sludge (OTBS). Sources such as polluted soils, petrochemical waste, sludge from refinery-wastewater plants, and others were used to obtain a collection of eight microorganisms, which were individually tested and characterized to analyze their degradative capabilities on different hydrocarbon families. After initial experiments using mixtures of these strains, we developed a consortium consisting of four microorganisms (three bacteria and one yeast) selected in the basis of their cometabolic effects, emulsification properties, colonization of oil components, and degradative capabilities. Although the specific contribution each of the former parameters makes is not clearly understood, the activity of the four-member consortium had a strong impact not only on linear alkane degradation (100%), but also on the degradation of cycloalkanes (85%), branched alkanes (44%), and aromatic and sulphur-aromatic compounds (31-55%). The effectiveness of this consortium was significantly superior to that obtained by individual strains, commercial inocula or an undefined mixture of culturable and non-culturable microorganisms obtained from OTBS-polluted soil. However, results were similar when another consortium of four microorganisms, previously isolated in the same OTBS-polluted soil, was assayed.