Comparative Life-Cycle Assessment of Aquifer Thermal Energy Storage Integrated with in Situ Bioremediation of Chlorinated Volatile Organic Compounds.

Due to the increasing need for sustainable energy and environmental quality in urban areas, the combination of aquifer thermal energy storage (ATES) and in situ bioremediation (ISB) has drawn much attention as it can deliver an integrated contribution to fulfill both demands. Yet, little is known about the overall environmental impacts of ATES-ISB. Hence, we applied a life-cycle assessment (LCA) to evaluate the environmental performance of ATES-ISB, which is also compared with the conventional heating and cooling system plus ISB alone (CHC + ISB). Energy supply via electricity is revealed as the primary cause of the environmental impacts, contributing 61.26% impacts of ATES-ISB and 72.91% impacts of CHC + ISB. Specifically, electricity is responsible for over 95% of water use, global warming potential, acidification potential, and respiratory inorganics, whereas the production of the biological medium for bioremediation causes more than 85% of the eco- and human toxicity impacts in both cases. The overall environmental impact of ATES-ISB is two times smaller than that of CHC + ISB. Sensitivity analysis confirms the importance of electricity consumption and electron donor production to the environmental impacts in both energy supply and bioremediation. Thus, future studies and practical applications seeking possible optimization of the environmental performances of ATES-ISB are recommended to focus more on these two essential elements, e.g., electricity and electron donor, and their related parameters. With the comprehensive LCA, insight is obtained for better characterizing the crucial factors as well as the relevant direction for future optimization research of the ATES-ISB system.

Combination of aquifer thermal energy storage and enhanced bioremediation: resilience of reductive dechlorination to redox changes.

To meet the demand for sustainable energy, aquifer thermal energy storage (ATES) is widely used in the subsurface in urban areas. However, contamination of groundwater, especially with chlorinated volatile organic compounds (CVOCs), is often being encountered. This is commonly seen as an impediment to ATES implementation, although more recently, combining ATES and enhanced bioremediation of CVOCs has been proposed. Issues to be addressed are the high water flow velocities and potential periodic redox fluctuation that accompany ATES. A column study was performed, at a high water flow velocity of 2 m/h, simulating possible changes in subsurface redox conditions due to ATES operation by serial additions of lactate and nitrate. The impacts of redox changes on reductive dechlorination as well as the microbial response of Dehalococcoides (DHC) were evaluated. The results showed that, upon lactate addition, reductive dechlorination proceeded well and complete dechlorination from cis-DCE to ethene was achieved. Upon subsequent nitrate addition, reductive dechlorination immediately ceased. Disruption of microorganisms' retention was also immediate and possibly detached DHC which preferred attaching to the soil matrix under biostimulation conditions. Initially, recovery of dechlorination was possible but required bioaugmentation and nutrient amendment in addition to lactate dosing. Repeated interruption of dechlorination and DHC activity by nitrate dosing appeared to be less easily reversible requiring more efforts for regenerating dechlorination. Overall, our results indicate that the microbial resilience of DHC in biosimulated ATES conditions is sensitive to redox fluctuations. Hence, combining ATES with bioremediation requires dedicated operation and monitoring on the aquifer geochemical conditions.

Biodegradation of cis-1,2-Dichloroethene in Simulated Underground Thermal Energy Storage Systems.

Underground thermal energy storage (UTES) use has showed a sharp rise in numbers in the last decades, with aquifer thermal energy storage (ATES) and borehole thermal energy storage (BTES) most widely used. In many urban areas with contaminated aquifers, there exists a desire for sustainable heating and cooling with UTES and a need for remediation. We investigated the potential synergy between UTES and bioremediation with batch experiments to simulate the effects of changing temperature and liquid exchange that occur in ATES systems, and of only temperature change occurring in BTES systems on cis-DCE reductive dechlorination. Compared to the natural situation (NS) at a constant temperature of 10 °C, both UTES systems with 25/5 °C for warm and cold well performed significantly better in cis-DCE (cis-1,2-dichloroethene) removal. The overall removal efficiency under mimicked ATES and BTES conditions were respectively 13 and 8.6 times higher than in NS. Inoculation with Dehalococcoides revealed that their initial presence is a determining factor for the dechlorination process. Temperature was the dominating factor when Dehalococcoides abundance was sufficient. Stimulated biodegradation was shown to be most effective in the mimicked ATES warm well because of the combined effect of suitable temperature, sustaining biomass growth, and regular cis-DCE supply.

Geochemical and microbiological characteristics during in situ chemical oxidation and in situ bioremediation at a diesel contaminated site.

While in situ chemical oxidation with persulfate has seen wide commercial application, investigations into the impacts on groundwater characteristics, microbial communities and soil structure are limited. To better understand the interactions of persulfate with the subsurface and to determine the compatibility with further bioremediation, a pilot scale treatment at a diesel-contaminated location was performed consisting of two persulfate injection events followed by a single nutrient amendment. Groundwater parameters measured throughout the 225 day experiment showed a significant decrease in pH and an increase in dissolved diesel and organic carbon within the treatment area. Molecular analysis of the microbial community size (16S rRNA gene) and alkane degradation capacity (alkB gene) by qPCR indicated a significant, yet temporary impact; while gene copy numbers initially decreased 1-2 orders of magnitude, they returned to baseline levels within 3 months of the first injection for both targets. Analysis of soil samples with sequential extraction showed irreversible oxidation of metal sulfides, thereby changing subsurface mineralogy and potentially mobilizing Fe, Cu, Pb, and Zn. Together, these results give insight into persulfate application in terms of risks and effective coupling with bioremediation.

Recovery of microbial diversity and activity during bioremediation following chemical oxidation of diesel contaminated soils.

To improve the coupling of in situ chemical oxidation and in situ bioremediation, a systematic analysis was performed of the effect of chemical oxidation with Fenton's reagent, modified Fenton's reagent, permanganate, or persulfate, on microbial diversity and activity during 8 weeks of incubation in two diesel-contaminated soils (peat and fill). Chemical oxidant and soil type affected the microbial community diversity and biodegradation activity; however, this was only observed following treatment with Fenton's reagent and modified Fenton's reagent, and in the biotic control without oxidation. Differences in the highest overall removal efficiencies of 69 % for peat (biotic control) and 59 % for fill (Fenton's reagent) were partially explained by changes in contaminant soil properties upon oxidation. Molecular analysis of 16S rRNA and alkane monooxygenase (alkB) gene abundances indicated that oxidation with Fenton's reagent and modified Fenton's reagent negatively affected microbial abundance. However, regeneration occurred, and final relative alkB abundances were 1-2 orders of magnitude higher in chemically treated microcosms than in the biotic control. 16S rRNA gene fragment fingerprinting with DGGE and prominent band sequencing illuminated microbial community composition and diversity differences between treatments and identified a variety of phylotypes within Alpha-, Beta-, and Gammaproteobacteria. Understanding microbial community dynamics during coupled chemical oxidation and bioremediation is integral to improved biphasic field application.

Impact of long-term diesel contamination on soil microbial community structure.

Microbial community composition and diversity at a diesel-contaminated railway site were investigated by pyrosequencing of bacterial and archaeal 16S rRNA gene fragments to understand the interrelationships among microbial community composition, pollution level, and soil geochemical and physical properties. To this end, 26 soil samples from four matrix types with various geochemical characteristics and contaminant concentrations were investigated. The presence of diesel contamination significantly impacted microbial community composition and diversity, regardless of the soil matrix type. Clean samples showed higher diversity than contaminated samples (P < 0.001). Bacterial phyla with high relative abundances in all samples included Proteobacteria, Firmicutes, Actinobacteria, Acidobacteria, and Chloroflexi. High relative abundances of Archaea, specifically of the phylum Euryarchaeota, were observed in contaminated samples. Redundancy analysis indicated that increased relative abundances of the phyla Chloroflexi, Firmicutes, and Euryarchaeota correlated with the presence of contamination. Shifts in the chemical composition of diesel constituents across the site and the abundance of specific operational taxonomic units (OTUs; defined using a 97% sequence identity threshold) in contaminated samples together suggest that natural attenuation of contamination has occurred. OTUs with sequence similarity to strictly anaerobic Anaerolineae within the Chloroflexi, as well as to Methanosaeta of the phylum Euryarchaeota, were detected. Anaerolineae and Methanosaeta are known to be associated with anaerobic degradation of oil-related compounds; therefore, their presence suggests that natural attenuation has occurred under anoxic conditions. This research underscores the usefulness of next-generation sequencing techniques both to understand the ecological impact of contamination and to identify potential molecular proxies for detection of natural attenuation.

Toluene biodegradation rates in unsaturated soil systems versus liquid batches and their relevance to field conditions.

Contaminant biodegradation in unsaturated soils may reduce the risks of vapor intrusion. However, the reported rates show large variability and are often derived from slurry experiments that are not representative of unsaturated conditions. Here, different laboratory setups are used to derive the biodegradation capacity of an unsaturated soil layer through which gaseous toluene migrates from the water table upwards. Experiments in static unsaturated soil microcosms at 6-30 % water-filled porosity (WFP) and unsaturated soil columns at 9, 14, and 27 % WFP were compared with liquid batches containing the same culture of Alicycliphilus denitrificans. The biodegradation rates for the liquid batches were orders of magnitude lower than for the other setups. Hence, liquid batches do not necessarily reflect optimal conditions for bacteria; either oxygen or toluene mass transfer at the cell scale or the absence of soil-water-air interfaces seemed to be limiting bacterial activity. For the column setup, the rates were limited by mass supply. The microcosm results could be described by apparent first-order biodegradation constants that increased with WFP or through a numerical model that included biodegradation as a first-order process taking place in the liquid phase only. The model liquid phase first-order rates varied between 6.25 and 20 h(-1) and were not related to the water content. Substrate availability was the primary factor limiting bioactivity, with evidence for physiological stress at the lowest water-filled porosity. The presented approach is useful to derive liquid phase biodegradation rates from experimental data and to include biodegradation in vapor intrusion models.

Biodegradation of aged diesel in diverse soil matrixes: impact of environmental conditions and bioavailability on microbial remediation capacity.

While bioremediation of total petroleum hydrocarbons (TPH) is in general a robust technique, heterogeneity in terms of contaminant and environmental characteristics can impact the extent of biodegradation. The current study investigates the implications of different soil matrix types (anthropogenic fill layer, peat, clay, and sand) and bioavailability on bioremediation of an aged diesel contamination from a heterogeneous site. In addition to an uncontaminated sample for each soil type, samples representing two levels of contamination (high and low) were also used; initial TPH concentrations varied between 1.6 and 26.6 g TPH/kg and bioavailability between 36 and 100 %. While significant biodegradation occurred during 100 days of incubation under biostimulating conditions (64.4-100 % remediation efficiency), low bioavailability restricted full biodegradation, yielding a residual TPH concentration. Respiration levels, as well as the abundance of alkB, encoding mono-oxygenases pivotal for hydrocarbon metabolism, were positively correlated with TPH degradation, demonstrating their usefulness as a proxy for hydrocarbon biodegradation. However, absolute respiration and alkB presence were dependent on soil matrix type, indicating the sensitivity of results to initial environmental conditions. Through investigating biodegradation potential across a heterogeneous site, this research illuminates the interplay between soil matrix type, bioavailability, and bioremediation and the implications of these parameters for the effectiveness of an in situ treatment.

Indene, indane and naphthalene in a mixture with BTEX affect aerobic compound biodegradation kinetics and indigenous microbial community development.

BTEX (benzene, toluene, ethylbenzene, xylene) are common pollutants often found in former gasworks sites together with some other contaminants like indene, indane and naphthalene (Ie, Ia, N). This study aimed to evaluate the inhibitory or stimulative substrate interactions between BTEX, and Ie, Ia, N during aerobic biodegradation. For this, batch bottles, containing originally anaerobic subsurface sediments, groundwater and indigenous microorganisms from a contaminated former gasworks site, were spiked with various substrate combinations (BTEX, BTEXIe, BTEXIa, BTEXN, BTEXIeIa, BTEXIeN, BTEXIaN, BTEXIeIaN). Subsequently concentrations were monitored over time. For the BTEXIeIaN mixture, initial concentrations were between 1 and 5 mg L-1, and all compounds were completely degraded by the microbial consortia within 39 days of incubation. The experimental data were fitted to a first order kinetic degradation model for interpretation of inhibition/stimulation between the compounds. Results showed that indene, indane, and naphthalene inhibited the degradation of benzene, toluene, ethylbenzene, o-xylene, with benzene being the most affected. M/p-xylene is the only compound whose biodegradation is stimulated by the presence of indene and indane (individually or mixed) but inhibited by the presence of naphthalene. 16S rRNA amplicon sequencing revealed differentiation in the microbial communities within the batches with different substrate mixtures, especially within the two microbial groups Micrococcaceae and Commamonaceae. Indene had more effect on the BTEX microbial community than indane or naphthalene and the presence of indene increased the relative abundance of Micrococcaceae family. In conclusion, co-presence of various pollutants leads to differentiation in degradation processes as well as in microbial community development. This sheds some light on the underlying reasons for that organic compounds present in mixtures in the subsurface of former gasworks sites are either recalcitrant or subjective towards biodegradation, and this understanding helps to further improve the bioremediation of such sites.

Spatial and temporal variation of metal concentrations in adult honeybees (Apis mellifera L.).

Honeybees (Apis mellifera L.) have great potential for detecting and monitoring environmental pollution, given their wide-ranging foraging behaviour. Previous studies have demonstrated that concentrations of metals in adult honeybees were significantly higher at polluted than at control locations. These studies focused at a limited range of heavy metals and highly contrasting locations, and sampling was rarely repeated over a prolonged period. In our study, the potential of honeybees to detect and monitor metal pollution was further explored by measuring the concentration in adult honeybees of a wide range of trace metals, nine of which were not studied before, at three locations in the Netherlands over a 3-month period. The specific objective of the study was to assess the spatial and temporal variation in concentration in adult honeybees of Al, As, Cd, Co, Cr, Cu, Li, Mn, Mo, Ni, Pb, Sb, Se, Sn, Sr, Ti, V and Zn. In the period of July-September 2006, replicated samples were taken at 2-week intervals from commercial-type bee hives. The metal concentration in micrograms per gram honeybee was determined by inductive coupled plasma-atomic emission spectrometry. Significant differences in concentration between sampling dates per location were found for Al, Cd, Co, Cr, Cu, Mn Sr, Ti and V, and significant differences in average concentration between locations were found for Co, Sr and V. The results indicate that honeybees can serve to detect temporal and spatial patterns in environmental metal concentrations, even at relatively low levels of pollution.

Degradation of 4-n-nonylphenol under nitrate reducing conditions.

Nonylphenol (NP) is an endocrine disruptor present as a pollutant in river sediment. Biodegradation of NP can reduce its toxicological risk. As sediments are mainly anaerobic, degradation of linear (4-n-NP) and branched nonylphenol (tNP) was studied under methanogenic, sulphate reducing and denitrifying conditions in NP polluted river sediment. Anaerobic bioconversion was observed only for linear NP under denitrifying conditions. The microbial population involved herein was further studied by enrichment and molecular characterization. The largest change in diversity was observed between the enrichments of the third and fourth generation, and further enrichment did not affect the diversity. This implies that different microorganisms are involved in the degradation of 4-n-NP in the sediment. The major degrading bacteria were most closely related to denitrifying hexadecane degraders and linear alkyl benzene sulphonate (LAS) degraders. The molecular structures of alkanes and LAS are similar to the linear chain of 4-n-NP, this might indicate that the biodegradation of linear NP under denitrifying conditions starts at the nonyl chain. Initiation of anaerobic NP degradation was further tested using phenol as a structure analogue. Phenol was chosen instead of an aliphatic analogue, because phenol is the common structure present in all NP isomers while the structure of the aliphatic chain differs per isomer. Phenol was degraded in all cases, but did not affect the linear NP degradation under denitrifying conditions and did not initiate the degradation of tNP and linear NP under the other tested conditions.

DNA-SIP identification of phenanthrene-degrading bacteria undergoing bioaugmentation and natural attenuation in petroleum-contaminated soil.

DNA-stable isotope probing (SIP) with 13C labeled phenanthrene (PHE) as substrate was used to identify specific bacterial degraders during natural attenuation (NA) and bioaugmentation (BA) in petroleum contaminated soil. BA, with the addition of a bacterial suspension mixture named GZ, played a significant role in PHE degradation with a higher PHE removal rate (∼90%) than that of NA (∼80%) during the first 3 days, and remarkably altered microbial communities. Of the five strains introduced in BA, only two genera, particularly, Ochrobactrum, Rhodococcus were extensively responsible for PHE-degradation. Six (Bacillus sp., Acinetobacter sp., Xanthomonas sp., Conexibacter sp., Acinetobacter sp. and Staphylococcus sp.) and seven (Ochrobactrum sp., Rhodococcus sp., Alkanindiges sp., Williamsia sp., Sphingobium sp., Gillisia sp. and Massilia sp.) bacteria responsible for PHE degradation were identified in NA and BA treatments, respectively. This study reports for the first time the association of Xanthomonas sp., Williamsia sp., and Gillisia sp. to PHE degradation.

Direct analysis of aromatic pollutants using a HPLC-FLD/DAD method for monitoring biodegradation processes.

Industrial discharges resulting in contaminated groundwater is a global environmental problem. For such contaminated groundwater cases, bioremediation is a cost efficient and environmentally friendly approach. The determination and quantification of these pollutants has gained great importance and researchers are currently seeking to develop labor extensive, accurate and reliable methods for evaluating their biodegradation process. In this study, a HPLC method was developed and optimized for the quantification of 11 industrial pollutants studied as two different mixtures: benzene, toluene, ethylbenzene, o, m/p-xylene, indane, indene, and naphthalene (mixture A) and benzene, monochlorobenzene, 1,2-dichlorobenzene, and 1,4-dichlorobenzene (mixture B). The method uses two different detectors: fluorescence detection and diode array. The fluorescence detector was used for mixture A to achieve lower quantification limits and to quantify separately o-xylene and indene due to them showing similar wavelength behaviors. The limit of detection was found to be between 2 and 70 µg L-1 for mixture A and 290 µg L-1 for mixture B. The limit of quantitation was between 6 and 210 µg L-1 for mixture A and 980 µg L-1 for mixture B, respectively. The novel part of this study is that aqueous samples can be directly measured with one-step sample preparation and it comes with other advantages such as low volumes of sampling from batch bottles and also avoidance of high cost, relative to other analytical techniques. Therefore, this analytical method aims to facilitate the quantification of various aromatic hydrocarbons in laboratory batch samples and can be used as a routine monitoring tool for biological degradation processes of these 11 prevalent contaminants.

Aerobic nonylphenol degradation and nitro-nonylphenol formation by microbial cultures from sediments.

Nonylphenol (NP) is an estrogenic pollutant which is widely present in the aquatic environment. Biodegradation of NP can reduce the toxicological risk. In this study, aerobic biodegradation of NP in river sediment was investigated. The sediment used for the microcosm experiments was aged polluted with NP. The biodegradation of NP in the sediment occurred within 8 days with a lag phase of 2 days at 30 degrees C. During the biodegradation, nitro-nonylphenol metabolites were formed, which were further degraded to unknown compounds. The attached nitro-group originated from the ammonium in the medium. Five subsequent transfers were performed from original sediment and yielded a final stable population. In this NP-degrading culture, the microorganisms possibly involved in the biotransformation of NP to nitro-nonylphenol were related to ammonium-oxidizing bacteria. Besides the degradation of NP via nitro-nonylphenol, bacteria related to phenol-degrading species, which degrade phenol via ring cleavage, are abundantly present.

Potential impact of hydrodynamic shear force in aquifer thermal energy storage on dissolved organic matter releasement: A vigorous shaking batch study.

The combination of bioremediation and aquifer thermal energy storage (ATES) has become attractive because of the possibility of solving environmental and energy problems simultaneously. While the impact of ATES on groundwater quality due to temperature change has received ample attention in literature, the effect of the greatly enhanced groundwater flow velocity on groundwater quality has not yet received sufficient scientific attention. To fill this gap in understanding, we conducted a simple yet straightforward experiment to illustrate the impact of hydrodynamic shear force due to the water flow by ATES on the release of dissolved organic matter, which can potentially be advantageous to bioremediation. Vigorous shaking conditions were applied to simulate the enhanced dynamics at the ATES well center and nearby. As the indicators of dissolved organic matter, COD and TOC concentrations were significantly impacted by shaking. COD increased from 5.4 mgO2/L to 36.3 mgO2/L during horizontal shaking. The maximum COD level was determined as 33.8 mgO2/L during orbital shaking, while the TOC level was growing from 6.7 to 28.7 mg C/L. Meanwhile, redox potential (with initial level -100 mV) was decreasing to -450 mV synchronously with the elevating COD and TOC level. Temperature was also revealed as a significant factor in the organic matter releasement. Microbial iron reduction was deemed to occur, yet sulfate reduction was not initiated during the whole experiment. Eventually, the structure of the soil-water matrix has been changed due to the extensive hydraulic and particle collisions, resulting in blackish appearance and thicker layer of fine particles. Overall, the findings advance our understanding of the role of the ATES-induced water flow in the subsurface biogeochemistry and give insight into the perspective of the combination of bioremediation and ATES. In general, an increase in dissolved organic matter can be expected due to the increased shear force at high flow conditions in the ATES system.

Tenax extraction as a tool to evaluate the availability of polybrominated diphenyl ethers, DTT, and DTT metabolites in sediments.

Solid-phase extraction with Tenax is one of the most used methods for determining the fraction of a pollutant that desorbs rapidly from sediment and thus is available for living beings. In the present study, this technique has been applied to sediment contaminated in the laboratory with polybrominated diphenyl ethers (PBDEs) and dichlorodiphenyltrichloroethane, dichlorodifenyldichloroethylene, and dichlorodifenyldichloroethane (generically, DDXs). The amount of chemical retained in sediment during the time of the experiment fit well with a three-phase exponential desorption model. The ratios between the rapidly desorbing fraction and the fraction desorbed during a fixed time (6 or 24 h) were calculated. The fraction desorbed in 6 h was lower than the rapidly desorbing fraction for most of PBDEs, whereas the fraction desorbed in 24 h exceeded the rapidly desorbing fraction for both groups of compounds. However, variability of these data suggests, when possible, a long time measure of desorption in order to achieve a more accurate estimation. Both the extent and the velocity of desorption were inversely related with the bromination degree and, consistently, with hydrophobicity and molecular size. In this way, low brominated PBDEs and DDXs showed a high availability relative to high brominated PBDEs.

Bioavailability and biodegradation of nonylphenol in sediment determined with chemical and bioanalysis.

The surfactant nonylphenol (NP) is an endocrine-disrupting compound that is widely spread throughout the environment. Although environmental risk assessments are based on total NP concentrations, only the bioavailable fraction possess an environmental risk. The present study describes the bioavailability and biodegradability of NP over time in contaminated river sediment of a tributary of the Ebro River in Spain. The bioavailable fraction was collected with Tenax TA(R) beads, and biodegradation was determined in aerobic batch experiments. The presence of NP was analyzed chemically using gas chromatography-mass spectrometry and indirectly as estrogenic potency using an in vitro reporter gene assay (ER(alpha)-luc assay). Of the total extractable NP in the sediment, 95%+/-1.5% (mean +/- standard error) desorbed quickly into the water phase. By aerobic biodegradation, the total extractable NP concentration and the estrogenic activity were reduced by 97%+/-0.5% and 94%+/-2%, respectively. The easily biodegradable fraction equals the potential bioavailable fraction. Only 43 to 86% of the estrogenic activity in the total extractable fraction, as detected in the ER(alpha)-luc assay, could be explained by the present NP concentration. This indicates that other estrogenic compounds were present and that their bioavailability and aerobic degradation were similar to that of NP. Therefore, we propose to use NP as an indicator compound to monitor estrogenicity of this Ebro River sediment. To what extent this conclusion holds for other river sediments depends on the composition of the contaminants and/or the nature of these sediments and requires further testing.

Combination of aquifer thermal energy storage and enhanced bioremediation: Biological and chemical clogging.

Interest in the combination concept of aquifer thermal energy storage (ATES) and enhanced bioremediation has recently risen due to the demand for both renewable energy technology and sustainable groundwater management in urban areas. However, the impact of enhanced bioremediation on ATES is not yet clear. Of main concern is the potential for biological clogging which might be enhanced and hamper the proper functioning of ATES. On the other hand, more reduced conditions in the subsurface by enhanced bioremediation might lower the chance of chemical clogging, which is normally caused by Fe(III) precipitate. To investigate the possible effects of enhanced bioremediation on clogging with ATES, we conducted two recirculating column experiments with differing flow rates (10 and 50mL/min), where enhanced biological activity and chemically promoted Fe(III) precipitation were studied by addition of lactate and nitrate respectively. The pressure drop between the influent and effluent side of the column was used as a measure of the (change in) hydraulic conductivity, as indication of clogging in these model ATES systems. The results showed no increase in upstream pressure during the period of enhanced biological activity (after lactate addition) under both flow rates, while the addition of nitrate lead to significant buildup of the pressure drop. However, at the flow rate of 10mL/min, high pressure buildup caused by nitrate addition could be alleviated by lactate addition. This indicates that the risk of biological clogging is relatively small in the investigated areas of the mimicked ATES system that combines enhanced bioremediation with lactate as substrate, and furthermore that lactate may counter chemical clogging.

Extraction and bioanalysis of the ecotoxicologically relevant fraction of contaminants in sediments.

Assessments of the risk connected to the contamination of soils and sediments should rely on a multidisciplinary approach based on both chemical and biological techniques (i.e., the sum of exposure and effects assessment). The dioxin-responsive, chemical-activated luciferase expression (DR-CALUX) bioassay is widely applied for evaluation of the toxicity of sediments after an exhaustive extraction of the contaminants, and results are used for risk assessment purposes. Approaches based on total extraction of contaminants do not take into account the importance of bioavailability and aging processes, thus leading to possible overestimations of risk. In the present work, an approach based on nonexhaustive extraction techniques in combination with an in vitro reporter gene assay was tested on sediment samples contaminated with dioxins, polycyclic aromatic hydrocarbons, polychlorinated biphenyls (PCBs), and other xenobiotics. Tenax and hydroxypropyl-beta-cyclodextrin (HPCD) extractions over time were carried out to determine the bioavailable fractions, whereas the residual fractions were determined by means of a microwave-assisted exhaustive extraction. For both fractions, contaminant concentrations were quantified by gas chromatography-mass spectrophotometry, and the toxic potency was determined by the DR-CALUX assay. Assessments of bioavailable fractions of PCBs by Tenax and HPCD gave comparable results and showed that after several years of aging, a considerable fraction (38-70% of the total content for different PCBs) is still available and ecotoxicologically relevant. Coupling of nonexhaustive extraction and bioanalyses leads to a more realistic and, generally, much lower estimated risk for the toxicity of the extracts as compared to commonly adopted exhaustive techniques.