Pump-and-treat (P&T) vs groundwater circulation wells (GCW): Which approach delivers more sustainable and effective groundwater remediation?

Pump-and-treat (P&T) is commonly used to remediate contaminated groundwater sites. The scientific community is currently engaged in a debate regarding the long-term effectiveness and sustainability of P&T for groundwater remediation. This work aims to provide a quantitative comparative analysis of the performance of an alternative system to traditional P&T, to support the development of sustainable groundwater remediation plans. Two industrial sites with unique geological frameworks and contamination with dense non-aqueous phase liquid (DNAPL) and arsenic (As) respectively, were selected for the study. At both locations, attempts were made for decades to clean up groundwater contamination by pump-and-treat. In response to persistently high levels of pollutants, groundwater circulation wells (GCWs) were installed to explore the possibility of accelerating the remediation process in unconsolidated and rock deposits. This comparative evaluation focuses on the different mobilization patterns observed, resulting variations in contaminant concentration, mass discharge, and volume of extracted groundwater. To facilitate the fusion of multi-source data, including geological, hydrological, hydraulic, and chemical information, and enable the continuous extraction of time-sensitive information, a geodatabase-supported conceptual site model (CSM) is utilized as a dynamic and interactive interface. This approach is used to assess the performance of GCW and P&T at the investigated sites. At Site 1, the GCW stimulated microbiological reductive dichlorination and mobilized significantly higher 1,2-DCE concentrations than P&T, despite recirculating a smaller volume of groundwater. At Site 2, As removal rate by GCW resulted generally higher than pumping wells. One conventional well mobilized higher masses of As in the early stages of P&T. This reflected the P&T's impact on accessible contaminant pools in early operational periods. P&T withdrew a significantly larger volume of groundwater than the GCW. The outcomes unveil the diverse contaminant removal behavior characterizing two distinct remediation strategies in different geological environments, revealing the dynamics and decontamination mechanisms that feature GCWs and P&T and emphasizing the limitations of traditional groundwater extraction systems in targeting aged pollution sources. GCWs have been shown to reduce remediation time, increase mass removal, and minimize the significant water consumption associated with P&T. These benefits pave the way for more sustainable groundwater remediation approaches in various hydrogeochemical scenarios.

Enhancing the Anaerobic Biodegradation of Petroleum Hydrocarbons in Soils with Electrically Conductive Materials.

Anaerobic bioremediation is a relevant process in the management of sites contaminated by petroleum hydrocarbons. Recently, interspecies electron transfer processes mediated by conductive minerals or particles have been proposed as mechanisms through which microbial species within a community share reducing equivalents to drive the syntrophic degradation of organic substrates, including hydrocarbons. Here, a microcosm study was set up to investigate the effect of different electrically conductive materials (ECMs) in enhancing the anaerobic biodegradation of hydrocarbons in historically contaminated soil. The results of a comprehensive suite of chemical and microbiological analyses evidenced that supplementing the soil with (5% w/w) magnetite nanoparticles or biochar particles is an effective strategy to accelerate the removal of selected hydrocarbons. In particular, in microcosms supplemented with ECMs, the removal of total petroleum hydrocarbons was enhanced by up to 50% relative to unamended controls. However, chemical analyses suggested that only a partial bioconversion of contaminants occurred and that longer treatment times would have probably been required to drive the biodegradation process to completion. On the other hand, biomolecular analyses confirmed the presence of several microorganisms and functional genes likely involved in hydrocarbon degradation. Furthermore, the selective enrichment of known electroactive bacteria (i.e., Geobacter and Geothrix) in microcosms amended with ECMs, clearly pointed to a possible role of DIET (Diet Interspecies Electron Transfer) processes in the observed removal of contaminants.

Microbiome Composition and Dynamics of a Reductive/Oxidative Bioelectrochemical System for Perchloroethylene Removal: Effect of the Feeding Composition.

Chlorinated solvents still represent an environmental concern that requires sustainable and innovative bioremediation strategies. This study describes the microbiome composition of a novel bioelectrochemical system (BES) based on sequential reductive/oxidative dechlorination for complete perchloroethylene (PCE) removal occurring in two separate but sequential chambers. The BES has been tested under various feeding compositions [i.e., anaerobic mineral medium (MM), synthetic groundwater (SG), and real groundwater (RG)] differing in presence of sulfate, nitrate, and iron (III). In addition, the main biomarkers of the dechlorination process have been monitored in the system under various conditions. Among them, Dehalococcoides mccartyi 16S rRNA and reductive dehalogenase genes (tceA, bvcA, and vcrA) involved in anaerobic dechlorination have been quantified. The etnE and etnC genes involved in aerobic dechlorination have also been quantified. The feeding composition affected the microbiome, in particular when the BES was fed with RG. Sulfuricurvum, enriched in the reductive compartment, operated with MM and SG, suggesting complex interactions in the sulfur cycle mostly including sulfur oxidation occurring at the anodic counter electrode (MM) or coupled to nitrate reduction (SG). Moreover, the known Mycobacterium responsible for natural attenuation of VC by aerobic degradation was found abundant in the oxidative compartment fed with RG, which was in line with the high VC removal observed (92 ± 2%). D. mccartyi was observed in all the tested conditions ranging from 8.78E + 06 (with RG) to 2.35E + 07 (with MM) 16S rRNA gene copies/L. tceA was found as the most abundant reductive dehalogenase gene in all the conditions explored (up to 2.46 E + 07 gene copies/L in MM). The microbiome dynamics and the occurrence of biomarkers of dechlorination, along with the kinetic performance of the system under various feeding conditions, suggested promising implications for the scale-up of the BES, which couples reductive with oxidative dechlorination to ensure the complete removal of highly chlorinated ethylene and mobile low-chlorinated by-products.

Contamination presence and dynamics at a polluted site: Spatial analysis of integrated data and joint conceptual modeling approach.

Contaminated sites are complex systems posing challenges for their characterization as both contaminant distribution and hydrogeological properties vary markedly at the metric scale, yet may extend over broad areas, with serious issues of spatial under-sampling in the space. Characterization with sufficient spatial resolution is thus, one of the main concerns and still open areas of research. To this end, the joint use of direct and indirect (i.e., geophysical) investigation methods is a very promising approach. This paper presents a case study aspiring to demonstrate the benefit of a multidisciplinary approach in the characterization of a hydrocarbon-contaminated site. Detailed multi-source data, collected via stratigraphic boreholes, laser-induced fluorescence (LIF) surveys, electrical resistivity tomography (ERT) prospecting, groundwater hydrochemical monitoring, and gas chromatography-mass spectrometry (GC-MS) analyses were compiled into an interactive big-data package for modeling activities. The final product is a comprehensive conceptual hydro-geophysical model overlapping multi-modality data and capturing hydrogeological and geophysical structures, as well as contamination distribution in space and dynamics in time. The convergence of knowledge in the joint model verifies the possibility of discriminating geophysical findings based on lithological features and contamination effects, unmasking the real characteristics of the pollutant, the contamination mechanisms, and the residual phase hydrocarbon sequestration linked to the hydrogeological dynamics and adopted remediation actions. The emerging conceptual site model (CSM), emphasizing the necessity of a large amount of multi-source data for its reliable, high-resolution reconstruction, appears as the necessary tool for the design of remedial actions, as well as for the monitoring of remediation performance.

A Polyhydroxybutyrate (PHB)-Biochar Reactor for the Adsorption and Biodegradation of Trichloroethylene: Design and Startup Phase.

In this work, polyhydroxy butyrate (PHB) and biochar from pine wood (PWB) are used in a mini-pilot scale biological reactor (11.3 L of geometric volume) for trichloroethylene (TCE) removal (80 mgTCE/day and 6 L/day of flow rate). The PHB-biochar reactor was realized with two sequential reactive areas to simulate a multi-reactive permeable barrier. The PHB acts as an electron donor source in the first "fermentative" area. First, the thermogravimetric (TGA) and differential scanning calorimetry (DSC) analyses were performed. The PHB-powder and pellets have different purity (96% and 93% w/w) and thermal properties. These characteristics may affect the biodegradability of the biopolymer. In the second reactive zone, the PWB works as a Dehalococcoides support and adsorption material since its affinity for chlorinated compounds and the positive effect of the "coupled adsorption and biodegradation" process has been already verified. A specific dechlorinating enriched culture has been inoculated in the PWB zone to realize a coupled adsorption and biodegradation process. Organic acids were revealed since the beginning of the test, and during the monitoring period the reductive dichlorination anaerobic pathway was observed in the first zone; no chlorinated compounds were detected in the effluent thanks to the PWB adsorption capacity.

Coupled Adsorption and Biodegradation of Trichloroethylene on Biochar from Pine Wood Wastes: A Combined Approach for a Sustainable Bioremediation Strategy.

Towards chlorinated solvents, the effectiveness of the remediation strategy can be improved by combining a biological approach (e.g., anaerobic reductive dechlorination) with chemical/physical treatments (e.g., adsorption). A coupled adsorption and biodegradation (CAB) process for trichloroethylene (TCE) removal is proposed in a biofilm-biochar reactor (BBR) to assess whether biochar from pine wood (PWB) can support a dechlorinating biofilm by combining the TCE (100 µM) adsorption. The BBR operated for eight months in parallel with a biofilm reactor (BR)-no PWB (biological process alone), and with an abiotic biochar reactor (ABR)-no dechlorinating biofilm (only an adsorption mechanism). Two flow rates were investigated. Compared to the BR, which resulted in a TCE removal of 86.9 ± 11.9% and 78.73 ± 19.79%, the BBR demonstrated that PWB effectively adsorbs TCE and slows down the release of its intermediates. The elimination of TCE was quantitative, with 99.61 ± 0.79% and 99.87 ± 0.51% TCE removal. Interestingly, the biomarker of the reductive dechlorination process, Dehalococcoides mccartyi, was found in the BRR (9.2 × 105 16S rRNA gene copies/g), together with the specific genes tceA, bvcA, and vcrA (8.16 × 106, 1.28 × 105, and 8.01 × 103 gene copies/g, respectively). This study suggests the feasibility of biochar to support the reductive dechlorination of D. mccartyi, opening new frontiers for field-scale applications.

Evaluation of the Bioelectrochemical Approach and Different Electron Donors for Biological Trichloroethylene Reductive Dechlorination.

Trichloroethylene (TCE) and more in general chlorinated aliphatic hydrocarbons (CAHs) can be removed from a contaminated matrix thanks to microorganisms able to perform the reductive dechlorination reaction (RD). Due to the lack of electron donors in the contaminated matrix, CAHs' reductive dechlorination can be stimulated by fermentable organic substrates, which slowly release molecular hydrogen through their fermentation. In this paper, three different electron donors constituted by lactate, hydrogen, and a biocathode of a bioelectrochemical cell have been studied in TCE dechlorination batch experiments. The batch reactors evaluated in terms of reductive dechlorination rate and utilization efficiency of the electron donor reported that the bio-electrochemical system (BES) showed a lower RD rate with respect of lactate reactor (51 ± 9 µeq/d compared to 98 ± 4 µeq/d), while the direct utilization of molecular hydrogen gave a significantly lower RD rate (19 ± 8 µeq/d), due to hydrogen low solubility in liquid media. The study also gives a comparative evaluation of the different electron donors showing the capability of the bioelectrochemical system to reach comparable efficiencies with a fermentable substrate without the use of other chemicals, 10.7 ± 3.3% for BES with respect of 3.5 ± 0.2% for the lactate-fed batch reactor. This study shows the BES capability of being an alternative at classic remediation approaches.

Control of Sulfate and Nitrate Reduction by Setting Hydraulic Retention Time and Applied Potential on a Membraneless Microbial Electrolysis Cell for Perchloroethylene Removal.

A membraneless microbial electrolysis cell (MEC) has been developed for perchloroethylene (PCE) removal through the reductive dechlorination reaction. The MEC consists of a tubular reactor of 8.24 L equipped with a graphite-granule working electrode which stimulates dechlorinating microorganisms while a graphite-granule cylindrical envelopment contained in a plastic mesh constituted the counter electrode of the MEC. Synthetic PCE-contaminated groundwater has been used as the feeding solution to test the nitrate and sulfate reduction reactions on the MEC performance at different hydraulic retention times (HRTs) (4.1, 1.8, and 1.2) and different cathodic potentials [-350, -450, and -650 mV vs standard hydrogen electrode (SHE)]. The HRT decrease from 4.1 to 1.8 d promoted a considerable increase in sulfate removal from 38 ± 11 to 113 ± 26 mg/Ld with a consequent current increase, while a shorter HRT of 1.2 d caused a partial inhibition of sulfate reduction with a consequent current decrease from -99 ± 3 to -52 ± 6 mA. Similarly, the cathodic potential investigation showed a direct correlation of current generation and sulfate removal in which the utilization of a cathodic potential of -350 mV versus SHE allowed for an 80% decrease in the sulfate removal rate with a consequent current decrease from -163 ± 7 to 41 ± 5 mA. The study showed the possibility to mitigate the energy consumption of the process by avoiding side reactions and current generation, through the selection of an appropriate HRT and applied cathodic potential.

Combined Strategies to Prompt the Biological Reduction of Chlorinated Aliphatic Hydrocarbons: New Sustainable Options for Bioremediation Application.

Groundwater remediation is one of the main objectives to minimize environmental impacts and health risks. Chlorinated aliphatic hydrocarbons contamination is prevalent and presents particularly challenging scenarios to manage with a single strategy. Different technologies can manage contamination sources and plumes, although they are usually energy-intensive processes. Interesting alternatives involve in-situ bioremediation strategies, which allow the chlorinated contaminant to be converted into non-toxic compounds by indigenous microbial activity. Despite several advantages offered by the bioremediation approaches, some limitations, like the relatively low reaction rates and the difficulty in the management and control of the microbial activity, can affect the effectiveness of a bioremediation approach. However, those issues can be addressed through coupling different strategies to increase the efficiency of the bioremediation strategy. This mini review describes different strategies to induce the reduction dechlorination reaction by the utilization of innovative strategies, which include the increase or the reduction of contaminant mobility as well as the use of innovative strategies of the reductive power supply. Subsequently, three future approaches for a greener and more sustainable intervention are proposed. In particular, two bio-based materials from renewable resources are intended as alternative, long-lasting electron-donor sources (e.g., polyhydroxyalkanoates from mixed microbial cultures) and a low-cost adsorbent (e.g., biochar from bio-waste). Finally, attention is drawn to novel bio-electrochemical systems that use electric current to stimulate biological reactions.

3D dynamic model empowering the knowledge of the decontamination mechanisms and controlling the complex remediation strategy of a contaminated industrial site.

Knowledge of the geology and hydrogeology of the polluted site emblematize a key requirement for environmental remediation, through assembling and synthesizing findings from various sources of physical evidence. In an increasingly virtual era, digital and geo-referenced metadata may serve as tools for collecting, merging, matching, and understanding multi-source information. The main goal of this paper is to emphasize the significance of a 3D hydrogeochemical model to the portrayal and the understanding of contamination dynamics and decontamination mechanisms at a highly contaminated industrial site. Some remediation measures are active on-site, due to the evidence-based presence of chlorinated solvents in groundwater. These are attributable to a slow-release source of pollutants in the saturated zone associated with very low permeability sediments. Therefore, in this research, a new technique for the remediation of secondary sources of dense non-aqueous phase liquid (DNAPL) contamination was investigated for the first time on a full-scale application. The combination of groundwater circulation wells (IEG-GCW®) and a continuous electron donor production device was set up to boost in situ bioremediation (ISB). A multi-phase approach was followed handling and releasing data during various remediation stages, from site characterization via pilot testing to full-scale remediation, thus allowing users to monitor, analyze, and manipulate information in 3D space-time. Multi-source and multi-temporal scenarios reveal the impact of ongoing hydraulic dynamics and depict the decontamination mechanisms in response to the interventions implemented over time, by quantifying the overall performance of the adopted strategies in terms of removal of secondary sources of pollution still active at the site.

Biochar from Pine Wood, Rice Husks and Iron-Eupatorium Shrubs for Remediation Applications: Surface Characterization and Experimental Tests for Trichloroethylene Removal.

Nowadays porous materials from organic waste, i.e., Biochar (BC), are receiving increased attention for environmental applications. This study adds information on three BCs that have undergone a number of studies in recent years. A Biochar from pine wood, one from rice husk and one from Eupatorium shrubs enriched with Iron, labelled as PWBC, RHBC and EuFeBC respectively, are evaluated for Trichloroethylene (TCE) removal from aqueous solution. Physical-chemical description is performed by SEM-EDS and BET analysis. The decrease of TCE over time follows a pseudo-second order kinetics with increased removal by the PWBC. Freundlich and Langmuir models well fit equilibrium test data. The optimized values of the maximum adsorbed amount, qmax (mg g-1), follows this order 109.41 PWBC > 30.35 EuFeBC > 21.00 RHBC. Fixed-bed columns are also carried out. Best performance is again achieved by PWBC, which operates for a higher number of pore volume, followed by EuFeBC and RHBC. Continuous testing confirms batch studies and makes it possible to evaluate the workability of materials in configurations closer to reality. Results are promising for potential environmental application. In particular, the characterization of several classes of contaminants opens the doors to possible uses in mixed contamination cases.

Understanding fungal potential in the mitigation of contaminated areas in the Czech Republic: tolerance, biotransformation of hexachlorocyclohexane (HCH) and oxidative stress analysis.

The study of the soil microbial community represents an important step in better understanding the environmental context. Therefore, biological characterisation and physicochemical integration are keys when defining contaminated sites. Fungi play a fundamental role in the soil, by providing and supporting ecological services for ecosystems and human wellbeing. In this research, 52 soil fungal taxa were isolated from in situ pilot reactors installed to a contaminated site in Czech Republic with a high concentration of hexachlorocyclohexane (HCH). Among the identified isolates, 12 strains were selected to evaluate their tolerance to different isomers of HCH by using specific indices (Rt:Rc; T.I.) and to test their potential in xenobiotic biotransformation. Most of the selected taxa was not significantly affected by exposure to HCH, underlining the elevated tolerance of all the tested fungal taxa, and different metabolic intermediates of HCH dechlorination were observed. The oxidative stress responses to HCH for two selected species, Penicillium simplicissimum and Trichoderma harzianum, were investigated in order to explore their toxic responses and to evaluate their potential functioning in bioremediation of contaminated environments. This research suggests that the isolated fungal species may provide opportunities for new eco-friendly, integrated and cost-effective solutions for environmental management and remediation, considering their efficient adaptation to stressful conditions.

Experimental and numerical evaluation of Groundwater Circulation Wells as a remediation technology for persistent, low permeability contaminant source zones.

Contaminants removal stoked inside low permeability zones of aquifers is one of the most important challenge of groundwater remediation process today. Low permeability layers can be considered persistent secondary sources of contamination because they release pollutants by molecular diffusion after primary source of contamination is reduced, causing long plum tails (Back-Diffusion). In this study, the Groundwater Circulation Well (GCW) system was investigated as an alternative remediation technology to the low efficient traditional pumping technologies to restore contaminated low permeability layers of aquifers. The GCW system creates vertical groundwater circulation cells by drawing groundwater through a screen of a multi-screen well and discharging it through another screen. The suitability of this technology to remediate contaminated low permeability zones was investigated by laboratory test and numerical simulations. The collected data were used to calibrate a model created to simulate the Back-Diffusion process and to evaluate the effect of different pumping technologies on the depletion time of that process. Results show that the efficiency of the GCW is dependent on the position and on the geometry of the low permeability zones, however the GCW system appears more suitable to restore contaminated low permeability layers of aquifers than the traditional pumping technology.

Microbial Community Changes in a Chlorinated Solvents Polluted Aquifer Over the Field Scale Treatment With Poly-3-Hydroxybutyrate as Amendment.

This study investigated the organohalide-respiring bacteria (OHRB) and the supporting microbial populations operating in a pilot scale plant employing poly-3-hydroxybutyrate (PHB), a biodegradable polymer produced by bacteria from waste streams, for the in situ bioremediation of groundwater contaminated by chlorinated solvents. The bioremediation was performed in ground treatment units, including PHB reactors as slow release source of electron donors, where groundwater extracted from the wells flows through before the re-infiltration to the low permeability zones of the aquifer. The coupling of the biological treatment with groundwater recirculation allowed to drastically reducing the contamination level and the remediation time by efficiently stimulating the growth of autochthonous OHRB and enhancing the mobilization of the pollutants. Quantitative PCR performed along the external treatment unit showed that the PHB reactor may efficiently act as an external incubator to growing Dehalococcoides mccartyi, known to be capable of fully converting chlorinated ethenes to innocuous end-products. The slow release source of electron donors for the bioremediation process allowed the establishment of a stable population of D. mccartyi, mainly carrying bvcA and vcrA genes which are implicated in the metabolic conversion of vinyl chloride to harmless ethene. Next generation sequencing was performed to analyze the phylogenetic diversity of the groundwater microbiome before and after the bioremediation treatment and allowed the identification of the microorganisms working closely with organohalide-respiring bacteria.

The bioelectric well: a novel approach for in situ treatment of hydrocarbon-contaminated groundwater.

Groundwater contamination by petroleum hydrocarbons (PHs) is a widespread problem which poses serious environmental and health concerns. Recently, microbial electrochemical technologies (MET) have attracted considerable attention for remediation applications, having the potential to overcome some of the limiting factors of conventional in situ bioremediation systems. So far, field-scale application of MET has been largely hindered by the limited availability of scalable system configurations. Here, we describe the 'bioelectric well' a bioelectrochemical reactor configuration, which can be installed directly within groundwater wells and can be applied for in situ treatment of organic contaminants, such as PHs. A laboratory-scale prototype of the bioelectric well has been set up and operated in continuous-flow regime with phenol as the model contaminant. The best performance was obtained when the system was inoculated with refinery sludge and the anode potentiostatically controlled at +0.2 V versus SHE. Under this condition, the influent phenol (25 mg l-1 ) was nearly completely (99.5 ± 0.4%) removed, with an average degradation rate of 59 ± 3 mg l-1 d and a coulombic efficiency of 104 ± 4%. Microbial community analysis revealed a remarkable enrichment of Geobacter species on the surface of the graphite anode, clearly pointing to a direct involvement of this electro-active bacterium in the current-generating and phenol-oxidizing process.

Metabolic synergies in the biotransformation of organic and metallic toxic compounds by a saprotrophic soil fungus.

The saprotrophic fungus Penicillium griseofulvum was chosen as model organism to study responses to a mixture of hexachlorocyclohexane (HCH) isomers (α-HCH, ß-HCH, γ-HCH, δ-HCH) and potentially toxic metals (vanadium, lead) in solid and liquid media. The P. griseofulvum FBL 500 strain was isolated from polluted soil containing high concentrations of HCH isomers and potentially toxic elements (Pb, V). Experiments were performed in order to analyse the tolerance/resistance of this fungus to xenobiotics and to shed further light on fungal potential in inorganic and organic biotransformations. The aim was to examine the ecological and bioremedial potential of this fungus verifying the presence of mechanisms that allow it to transform HCH isomers and metals under different extreme test conditions. To our knowledge, this work is the first to provide evidence on the biotransformation of HCH mixtures, in combination with toxic metals, by a saprotrophic non-white-rot fungus and on the metabolic synergies involved.

The "Oil-Spill Snorkel": an innovative bioelectrochemical approach to accelerate hydrocarbons biodegradation in marine sediments.

This study presents the proof-of-concept of the "Oil-Spill Snorkel": a novel bioelectrochemical approach to stimulate the oxidative biodegradation of petroleum hydrocarbons in sediments. The "Oil-Spill Snorkel" consists of a single conductive material (the snorkel) positioned suitably to create an electrochemical connection between the anoxic zone (the contaminated sediment) and the oxic zone (the overlying O2-containing water). The segment of the electrode buried within the sediment plays a role of anode, accepting electrons deriving from the oxidation of contaminants. Electrons flow through the snorkel up to the part exposed to the aerobic environment (the cathode), where they reduce oxygen to form water. Here we report the results of lab-scale microcosms setup with marine sediments and spiked with crude oil. Microcosms containing one or three graphite snorkels and controls (snorkel-free and autoclaved) were monitored for over 400 days. Collectively, the results of this study confirmed that the snorkels accelerate oxidative reactions taking place within the sediment, as documented by a significant 1.7-fold increase (p = 0.023, two-tailed t-test) in the cumulative oxygen uptake and 1.4-fold increase (p = 0.040) in the cumulative CO2 evolution in the microcosms containing three snorkels compared to snorkel-free controls. Accordingly, the initial rate of total petroleum hydrocarbons (TPH) degradation was also substantially enhanced. Indeed, while after 200 days of incubation a negligible degradation of TPH was noticed in snorkel-free controls, a significant reduction of 12 ± 1% (p = 0.004) and 21 ± 1% (p = 0.001) was observed in microcosms containing one and three snorkels, respectively. Although, the "Oil-Spill Snorkel" potentially represents a groundbreaking alternative to more expensive remediation options, further research efforts are needed to clarify factors and conditions affecting the snorkel-driven biodegradation processes and to identify suitable configurations for field applications.

Biotransformation of ß-hexachlorocyclohexane by the saprotrophic soil fungus Penicillium griseofulvum.

ß-Hexachlorocyclohexane (ß-HCH) is a persistent organic pollutant (POP) of global concern with potentially toxic effects on humans and ecosystems. Fungal tolerance and biotransformation of toxic substances hold considerable promise in environmental remediation technologies as many fungi can tolerate extreme environmental conditions and possess efficient extracellular degradative enzymes with relatively non-specific activities. In this research, we have investigated the potential of a saprotrophic soil fungus, Penicillium griseofulvum Dierckx, isolated from soils with high concentrations of isomers of hexachlorocyclohexane, to biotransform ß-HCH, the most recalcitrant isomer to microbial activity. The growth kinetics of the fungus were characterized after growth in stirred liquid Czapek-Dox medium. It was found that P. griseofulvum was able to grow in the presence of 1 mg L(-1) ß-HCH and in stressful nutritional conditions at different concentrations of sucrose in the medium (0 and 5 g L(-1)). The effects of ß-HCH and the toluene, used as a solvent for ß-HCH addition, on P. griseofulvum were investigated by means of a Phenotype MicroArray™ technique, which suggested the activation of certain metabolic pathways as a response to oxidative stress due to the presence of the xenobiotics. Gas chromatographic analysis of ß-HCH concentration confirmed biodegradation of the isomer with a minimum value of ß-HCH residual concentration of 18.6%. The formation of benzoic acid derivatives as dead-end products of ß-HCH biotransformation was observed and this could arise from a possible biodegradation pathway for ß-HCH with important connections to fungal secondary metabolism.

Relevance of side reactions in anaerobic reductive dechlorination microcosms amended with different electron donors.

In this study we examined the relative importance of side reactions, i.e. the formation of volatile fatty acids (VFA), and the reduction of alternative electron acceptors (nitrate, sulfate, Fe(III)), in enhanced dechlorination microcosms, amended with different electron donors, namely lactate, butyrate, and H(2)+acetate mixture. Dechlorination reactions proceeded at low rates and consequently, nearly all of the reducing equivalents (over 99%) available from electron donor consumption were channeled to (side) reactions other than dechlorination. The relevance of these side reactions was more evident with lactate which was consumed at higher rate than other electron donors. Correspondingly, high levels of VFA and soluble Fe(II) accumulated in the supernatant of lactate-amended microcosms. Ecotoxicity experiments (Lepidium sativum germination tests) also indicated that the supernatant was much more toxic/inhibitory than that of other microcosms. Among the electron donors tested, the H(2)+acetate mixture, yielded the most promising results in terms of extent of dechlorination, negligible accumulation of by-products, and residual groundwater toxicity. Fluorescent In situ Hybridization analysis (FISH) confirmed that H(2)+acetate-amended microcosms were dominated by Dehalococcoides spp., while a higher biodiversity was observed in the cultures fed with lactate or butyrate. Overall, the average amount of donor that was required for the removal of 1micromol of chloride from the contaminant differed greatly among the donors, namely 2.13meq/micromolCl(-) for lactate, 1.01meq/micromolCl(-) for butyrate, and 0.39meq/micromolCl(-) for H(2)+acetate mixture. Interestingly, dechlorinating activity was observed under sulfate-reducing conditions; this suggests that it may not be necessary to deplete the sulfate from the groundwater, for instance by supplying an excess electron donor, in order to achieve substantial dechlorination. Finally, in this study we found that the pathway of anaerobic lactate degradation shifted from the production of acetate and H(2) during active sulfate reduction to acetate and propionate upon sulfate depletion. Energetic considerations that support this finding were presented.

Assessment of natural or enhanced in situ bioremediation at a chlorinated solvent-contaminated aquifer in Italy: a microcosm study.

A microcosm study was used to assess the potential for in situ natural or enhanced bioremediation at a chloroethane- (i.e., tetrachloroethane, TeCA) and chloroethene-contaminated (i.e., tetrachloroethene, PCE; trichloroethene, TCE) groundwater in Northern Italy. All the live microcosms were positive for dechlorination, indicating the presence of an active native dechlorinating population in the subsurface. All the tested electron donors (i.e., yeast extract, lactate, butyrate, hydrogen) promoted enhanced dechlorination of chlorinated contaminants. Lactate- and butyrate-amended microcosms performed the best, and also dechlorinated the solvents past cis-dichloroethene (cis-DCE). The microcosm bioaugmented with a PCE-dechlorinating mixed culture containing Dehalococcoides spp. dechlorinated groundwater contaminants to DCE, vinyl chloride (VC), and ethene (ETH). In conclusion, results from this microcosm study indicate the potential for enhancing full dechlorination at the contaminated site, through a proper addition of a suitable electron donor (e.g., lactate or butyrate) and/or through bioaugmentation with a Dehalococcoides-containing culture.