Influence of combined abiotic/biotic factors on decay of P. aeruginosa and E. coli in Rhine River water.

Understanding the dynamic change in abundance of both fecal and opportunistic waterborne pathogens in urban surface water under different abiotic and biotic factors helps the prediction of microbiological water quality and protection of public health during recreational activities, such as swimming. However, a comprehensive understanding of the interaction among various factors on pathogen behavior in surface water is missing. In this study, the effect of salinity, light, and temperature and the presence of indigenous microbiota, on the decay/persistence of Escherichia coli and Pseudomonas aeruginosa in Rhine River water were tested during 7 days of incubation with varying salinity (0.4, 5.4, 9.4, and 15.4 ppt), with light under a light/dark regime (light/dark) and without light (dark), temperature (3, 12, and 20 °C), and presence/absence of indigenous microbiota. The results demonstrated that light, indigenous microbiota, and temperature significantly impacted the decay of E. coli. Moreover, a significant (p<0.01) four-factor interactive impact of these four environmental conditions on E. coli decay was observed. However, for P. aeruginosa, temperature and indigenous microbiota were two determinate factors on the decay or growth. A significant three-factor interactive impact between indigenous microbiota, temperature, and salinity (p<0.01); indigenous microbiota, light, and temperature (p<0.01); and light, temperature, and salinity (p<0.05) on the decay of P. aeruginosa was found. Due to these interactive effects, caution should be taken when predicting decay/persistence of E. coli and P. aeruginosa in surface water based on a single environmental condition. In addition, the different response of E. coli and P. aeruginosa to the environmental conditions highlights that E. coli monitoring alone underestimates health risks of surface water by non-fecal opportunistic pathogens, such as P. aeruginosa. KEY POINTS: Abiotic and biotic factors interactively affect decay of E. coli and P. aeruginosa E.coli and P.aeruginosa behave significantly different under the given conditions Only E. coli as an indicator underestimates the microbiological water quality.

Electrochemical removal of amphoteric ions.

Several harmful or valuable ionic species present in seawater, brackish water, and wastewater are amphoteric, weak acids or weak bases, and, thus, their properties depend on local water pH. Effective removal of these species can be challenging for conventional membrane technologies, necessitating chemical dosing of the feedwater to adjust pH. A prominent example is boron, which is considered toxic in high concentrations and often requires additional membrane passes to remove during seawater desalination. Capacitive deionization (CDI) is an emerging membraneless technique for water treatment and desalination, based on electrosorption of salt ions into charging microporous electrodes. CDI cells show strong internally generated pH variations during operation, and, thus, CDI can potentially remove pH-dependent species without chemical dosing. However, development of this technique is inhibited by the complexities inherent to the coupling of pH dynamics and ion properties in a charging CDI cell. Here, we present a theoretical framework predicting the electrosorption of pH-dependent species in flow-through electrode CDI cells. We demonstrate that such a model enables insight into factors affecting species electrosorption and conclude that important design rules for such systems are highly counterintuitive. For example, we show both theoretically and experimentally that for boron removal, the anode should be placed upstream and the cathode downstream, an electrode order that runs counter to the accepted wisdom in the CDI field. Overall, we show that to achieve target separations relying on coupled, complex phenomena, such as in the removal of amphoteric species, a theoretical CDI model is essential.

Bioaugmentation has temporary effect on anaerobic pesticide biodegradation in simulated groundwater systems.

Groundwater is the most important source for drinking water in The Netherlands. Groundwater quality is threatened by the presence of pesticides, and biodegradation is a natural process that can contribute to pesticide removal. Groundwater conditions are oligotrophic and thus biodegradation can be limited by the presence and development of microbial communities capable of biodegrading pesticides. For that reason, bioremediation technologies such as bioaugmentation (BA) can help to enhance pesticide biodegradation. We studied the effect of BA using enriched mixed inocula in two column bioreactors that simulate groundwater systems at naturally occurring redox conditions (iron and sulfate-reducing conditions). Columns were operated for around 800 days, and two BA inoculations (BA1 and BA2) were conducted in each column. Inocula were enriched from different wastewater treatment plants (WWTPs) under different redox-conditions. We observed a temporary effect of BA1, reaching 100% removal efficiency of the pesticide 2,4-D after 100 days in both columns. In the iron-reducing column, 2,4-D removal was in general higher than under sulfate-reducing conditions demonstrating the influence of redox conditions on overall biodegradation. We observed a temporary shift in microbial communities after BA1 that is relatable to the increase in 2,4-D removal efficiency. After BA2 under sulfate-reducing conditions, 2,4-D removal efficiency decreased, but no change in the column microbial communities was observed. The present study demonstrates that BA with a mixed inoculum can be a valuable technique for improving biodegradation in anoxic groundwater systems at different redox-conditions.

The complexities of decision-making in food waste valorization: A critical review.

The efficient utilization of food waste (FW) resources through Food Waste Valorization (FWV) has received increasing attention in recent years. Various decision-making studies have been undertaken to facilitate FWV implementation, such as the studies on decision-making framework and FWV technology assessment. Food waste hierarchy is a widely discussed framework in FW management, but it was found too simplified and does not always contribute positively to environmental sustainability. Moreover, decision-making studies in FWV often focus on specific aspects of the food system and employ distinctive decision-making approaches, making it difficult to compare the results from different studies. Therefore, our literature review is conducted to provide a comprehensive understanding of FWV decision-making. This study identifies what decisions are needed, and three levels of decisions are revealed: system-level, FW stream-level, and FWV option-level. The assessment approaches and criteria used to support decision-making in FWV are also collected and analyzed. Building upon these findings, an hourglass model is synthesized to provide a holistic illustration of decision-making in FWV. This study untangles the complexities of FWV decision-making and sheds light on the limitations of current studies. We anticipate this study will make more people realize that FWV is a multidisciplinary issue and requires the collective participation of researchers, practitioners, policymakers, and consumers. Such collective engagement is essential to effectively address practical challenges and propel the transition of the current food system toward a more resource-efficient paradigm.

Seasonal variation in antibiotic resistance genes and bacterial phenotypes in swine wastewater during three-chamber anaerobic pond treatment.

Antibiotic resistance is a global public health concern. Antibiotic usage in pigs makes swine wastewater (SW) a reservoir for antibiotic resistance genes (ARGs). SW is usually stored and treated in a three-chamber anaerobic pond (3-CAP) in medium and small pig farms in northern China. However, the yet unexplored presence of ARGs in SW during 3-CAP treatment may result in ARGs spreading into the environment if farmers apply SW to farmland as a liquid organic fertilizer. This study investigated the profiles of and changes in ARGs in SW during its treatment in 3-CAP over four seasons and analyzed the correlation between ARGs and bacterial phenotypes, along with the physicochemical parameters of the water. The results revealed that ARG abundance decreased considerably after 3-CAP treatment in April (47%), October (47%), and December (62%) but increased in May (43%) and August (73%). The ARG copies in the influent and other SW samples increased significantly from 107 copies/mL in April to 109 copies/mL in October and were maintained in December. The increase in ARG abundance was not as rapid as the growth of the bacterial population, resulting in lower relative abundance in October and December. Bacterial communities possessed more sul1 and tetM genes, which were also positively correlated with mobile genetic elements. After the 3-CAP treatment, 16% of antibiotics and 60% of heavy metals were removed, and both had a weak correlation with ARGs. Predicted phenotypes showed that gram-positive (G+) and gram-negative (G-) bacteria have different capacities for carrying ARGs. G+ bacteria carry more ARGs than G- bacteria. This study revealed the persistence of ARGs in SW after 3-CAP treatment over different seasons. Applying SW in the proper month will mitigate ARG dissemination to the environment.

Effect of electrolyte composition on electrochemical oxidation: Active sulfate formation, benzotriazole degradation, and chlorinated by-products distribution.

Electrochemical oxidation is an effective technique for treating persistent organic pollutants, which are hardly removed in conventional wastewater treatment plants. Sulfate and chloride salts commonly used and present in natural wastewater influence the electrochemical degradation process. In this study, the effect of electrolyte composition on the active sulfate species (SO4●⁻ and S2O82⁻) formation, benzotriazole degradation-a model organic compound, and chlorinated by-products distribution have been investigated while using a boron-doped diamond (BDD) anode. Different Na2SO4:NaNO3 and Na2SO4:NaCl ratios with constant conductivity of 10 mS/cm were used in the experiments and applied anode potential was kept constant at 4.3 V vs. Ag/AgCl. The electrogenerated SO4●⁻ and S2O82⁻ formation were faster in 10:1 and 2:1 Na2SO4:NaNO3 ratios than in the 1:0 ratio. The ●OH-mediated SO4●⁻ production has prevailed in 10:1 and 2:1 ratios. However, ●OH-mediated SO4●⁻ production has hindered the 1:0 ratio due to excess chemisorption of SO42⁻ on the BDD anode. Similarly, the faster benzotriazole degradation, mineralization, and lowest energy consumption were achieved in the 10:1 Na2SO4:NaNO3 and Na2SO4:NaCl ratio. Besides, chlorinated organic by-product concentration (AOX) was lower in the 10:1 Na2SO4:NaCl ratio but increased with the increasing chloride ratio in the electrolyte. LC-MS analysis shows that several chlorinated organic transformation products were produced in 0:1 to 2:1 ratio, which was not found in the 10:1 Na2SO4:NaCl ratio. A comparatively higher amount of ClO4⁻ was formed in the 10:1 ratio than in 2:1 to 0:1 ratio. This ClO4⁻ formation train evidence the effective ●OH generation in a sulfate-enriched condition because the ClO4⁻ formation is positively correlated to ●OH concentration. Overall results show that sulfate-enriched electrolyte compositions are beneficial for electrochemical oxidation of biorecalcitrant organic pollutants.

Mathematically formulated key performance indicators for design and evaluation of treatment trains for resource recovery from urban wastewater.

While urban wastewater infrastructure is aging and no longer adequate, climate change and sustainability are urging the transition from pollution management to resource recovery. Lacking evidence-based quantitative evaluation of the potential benefits and consequences of resource recovery from wastewater hinders the negotiation amongst stakeholders and slows down the transition. This study proposes mathematical formulations for technical, environmental, economic, and social key performance indicators (KPIs) that can be used to quantify the benefits and the risks of resource recovery. The proposed formulations are derived from the literature and validated with stakeholders. Each KPI is mathematically formulated at treatment train level by considering: (1) the characteristics of individual unit processes (UPs) in the treatment train (TT), (2) the context in which the TT is installed, and (3) the resources to be recovered. The mathematical formulations of the KPIs proposed in this study enable a transparent, consistent and informative evaluation of existing treatment trains, as well as support the (computer aided) design of new ones. This could aid the transition from urban wastewater treatment to resource recovery from urban wastewater.

Pilot-scale hybrid constructed wetlands for the treatment of cooling tower water prior to its desalination and reuse.

Cooling towers are responsible for a large part of the industrial fresh water withdrawal, and the reuse of cooling tower water (CTW) effluents can strongly lower industrial fresh water footprints. CTW requires desalination prior to being reused, but various CTW components, such as total organic carbon (TOC), conditioning chemicals and total suspended solids (TSS) hamper physico-chemical desalination technologies and need to be removed from the CTW. A cost-efficient and robust pre-treatment is thus required, which can be provided by constructed wetlands (CWs). The present study is the first study that determined the CTW pre-treatment efficiency of hybrid-CWs and the impact of winter season and biocides in the CTW on the pre-treatment efficiency. The most efficient CW flow type and dominant removal mechanisms for CW components hampering physico-chemical desalination were determined. Subsurface flow CWs removed PO43-, TSS and TOC as a result of adsorption and filtration. Vertical subsurface flow CWs (VSSF-CW) excelled in the removal of benzotriazole as a result of aerobic biodegradation. Horizontal subsurface flow CWs (HSSF-CW) allowed the denitrification of NO3- due to their anaerobic conditions. Open water CWs (OW-CWs) did not contribute to the removal of components that hamper physico-chemical desalination technologies, but do provide water storage options and habitat. The biological removal processes in the different CW flow types were negatively impacted by the winter season, but were not impacted by concentrations of the biocides glutaraldehyde and DBNPA that are relevant in practice. For optimal pre-treatment, a hybrid-CW, consisting of an initial VSSF-CW followed by an OW-CW and HSSF-CW is recommended. Future research should focus on integrating the hybrid-CW with a desalination technology, e.g. reverse osmosis, electrodialysis or capacitive ionization, to produce water that meets the requirements for use as cooling water and allow the reuse of CTW in the cooling tower itself.

Estimation of the water cycle related to shale gas production under high data uncertainties: Dutch perspective.

The potential water demand for fracturing fluids along with the possible flowback and produced water production is assessed for the Dutch Posidonia shale. Total water demand estimated for 25 years of the field development using historic data from the U.S. plays varies between 12.2 and 36.9 Mm3. The maximal annual water consumption of 0.95-2.88 Mm3 is expected in the peak years of shale gas production. These figures are much lower than the availability of any potential water sources, which include drinking water, fresh and brackish groundwater, river water, effluents of wastewater treatment plants (WWTP) and sea water. River water is considered the most promising water source for fracturing fluids in the Dutch Posidonia shale based on its availability (>6·104 Mm3/year) and quality (only bacterial composition needs to be controlled). Total wastewater production for the whole period of the field development is estimated between 6.6 and 48.0 Mm3. Wastewater recycling can cover significant part of the source water demand for fracturing fluid. However, high mineral content of the wastewater as well as temporal and spatial discrepancies between wastewater production and water demand will form obstacles for wastewater recycling. The assessment framework developed in this study may be applied for other shale gas fields with high uncertainties regarding subsurface properties, connate formation water characteristics and future legislative framework.

Organic Pollutants in Shale Gas Flowback and Produced Waters: Identification, Potential Ecological Impact, and Implications for Treatment Strategies.

Organic contaminants in shale gas flowback and produced water (FPW) are traditionally expressed as total organic carbon (TOC) or chemical oxygen demand (COD), though these parameters do not provide information on the toxicity and environmental fate of individual components. This review addresses identification of individual organic contaminants in FPW, and stresses the gaps in the knowledge on FPW composition that exist so far. Furthermore, the risk quotient approach was applied to predict the toxicity of the quantified organic compounds for fresh water organisms in recipient surface waters. This resulted in an identification of a number of FPW related organic compounds that are potentially harmful namely those compounds originating from shale formations (e.g., polycyclic aromatic hydrocarbons, phthalates), fracturing fluids (e.g., quaternary ammonium biocides, 2-butoxyethanol) and downhole transformations of organic compounds (e.g., carbon disulfide, halogenated organic compounds). Removal of these compounds by FPW treatment processes is reviewed and potential and efficient abatement strategies are defined.

Metabolism of Ibuprofen by Phragmites australis: Uptake and Phytodegradation.

This study explores ibuprofen (IBP) uptake and transformation in the wetland plant species Phragmites australis and the underlying mechanisms. We grew P. australis in perlite under greenhouse conditions and treated plants with 60 µg/L of IBP. Roots and rhizomes (RR), stems and leaves (SL), and liquid samples were collected during 21 days of exposure. Results show that P. australis can take up, translocate, and degrade IBP. IBP was completely removed from the liquid medium after 21 days with a half-life of 2.1 days. IBP accumulated in RR and was partly translocated to SL. Meanwhile, four intermediates were detected in the plant tissues: hydroxy-IBP, 1,2-dihydroxy-IBP, carboxy-IBP and glucopyranosyloxy-hydroxy-IBP. Cytochrome P450 monooxygenase was involved in the production of the two hydroxy intermediates. We hypothesize that transformation of IBP was first catalyzed by P450, and then by glycosyltransferase, followed by further storage or metabolism in vacuoles or cell walls. No significant phytotoxicity was observed based on relative growth of plants and stress enzyme activities. In conclusion, we demonstrated for the first time that P. australis degrades IBP from water and is therefore a suitable species for application in constructed wetlands to clean wastewater effluents containing IBP and possibly also other micropollutants.

Metallic nanoparticles: microbial synthesis and unique properties for biotechnological applications, bioavailability and biotransformation.

The impact of nanotechnology in all areas of science and technology is evident. The expanding availability of a variety of nanostructures with properties in the nanometer size range has sparked widespread interest in their use in biotechnological systems, including the field of environmental remediation. Nanomaterials can be used as catalysts, adsorbents, membranes, water disinfectants and additives to increase catalytic activity and capability due to their high specific surface areas and nanosize effects. Thus, nanomaterials appear promising for new effective environmental technologies. Definitely, nanotechnology applications for site remediation and wastewater treatment are currently in research and development stages, and new innovations are underway. The synthesis of metallic nanoparticles has been intensively developed not only due to its fundamental scientific interest but also for many technological applications. The use of microorganisms in the synthesis of nanoparticles is a relatively new eco-friendly and promising area of research with considerable potential for expansion. On the other hand, chemical synthesis occurs generally under extreme conditions (e.g. pH, temperature) and also chemicals used may have associated environmental and human health impacts. This review is an overview of current research worldwide on the use of microorganisms during the biosynthesis of metallic nanoparticles and their unique properties that make them good candidates for many applications, including in biotechnology.

Effect of granular activated carbon concentration on the content of organic matter and salt, influencing E. coli activity and survival in fluidized bed disinfection reactor.

Granular activated carbon (GAC) is used in water treatment systems, typically to remove pollutants such as natural organic matter, volatile organic compounds, chlorine, taste, and odor. GAC is also used as a key component of a new technology that combines a fluidized bed reactor with radio frequency electric fields for disinfection. So far, the effects of GAC on bacteria in these fluidized bed reactors are unclear. This paper describes a systematic study of the physico-chemical changes in five microbial media compositions caused by different concentrations (23-350 g/L) of GAC, and the effects of these physico-chemical changes on the metabolic activity and survival of a model microorganism (Escherichia coli YMc10) in a fluidized bed reactor. The chemical adsorption taking place in suspensions with specific GAC changed nutritional, osmotic, and pH conditions in the investigated microbial media (LB, diluted LB, PBS, diluted PBS, and tap water), leading to a decay of the metabolic activity and survival of E. coli. Especially media that are poor in organic and mineral compounds (e.g., PBS) with suspended GAC showed a concentration decay of 3.5 Log CFU/mL E. coli after 6 h. Organic compounds depletion and severe pH variation were enhanced in the presence of higher GAC concentrations.

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.

Hydroponic materials improve organic micropollutant removal in vertical flow constructed wetlands treating wastewater.

Unconventional substrata like activated carbon or clay beads can enhance micropollutant removal in constructed wetlands. However, hydroponic materials widely used in horticulture have not yet been investigated for their potential to remove micropollutants. In addition, potential effect of plant species other than reeds on micropollutant removal has not been sufficiently investigated. Therefore, a nature-based, post-treatment technology called improved vertical flow constructed wetlands (CW) with hydroponic (H) materials (CWH) was designed by employing cocopeat and mineral with ornamental plant species syngonium and periwinkle. A mesocosm CWH system was tested in a climate-controlled greenhouse for 550 days for its potential to remove frequently found micropollutants in wastewater, namely sulfamethoxazole, trimethoprim, diclofenac, erythromycin, carbamazepine, pyrimethanil, tebuconazole, pymetrozine, atrazine and DEET from wastewater effluent. The main focus was to understand the contribution of sorption, microbial degradation and phytoremediation on the removal of those micropollutants. It was found that cocopeat showed a capacity for sorbing micropollutants, ranging between 80 and 99% of the compounds added while less than 10% sorption was observed for mineral wool. Additionally moderate to high biological removal (25-60 µg of compound/kg dry weight of substratum/day) for most of the studied compounds was observed in all the cocopeat biotic groups. Furthermore, it could be stated that plants appear not to be an important factor for micropollutant removal. The observed differences in removal between the cocopeat and mineral wool systems could be explained by the difference in physico-chemical properties of the substrata, where cocopeat has a higher water holding capacity, moisture content, nutrient and organic matter content, and a higher intraparticle porosity and surface area. This study revealed notable removal of persistent and mobile micropollutants in cocopeat CWH, namely carbamazepine (80-86%) and diclofenac (97-100%). These results demonstrate the potential beneficial use of hydroponic materials as substratum in more advanced constructed wetlands designed to remove micropollutants.

Multi-component ion equilibria and transport in ion-exchange membranes.

At the interface between an ion-exchange membrane and a multi-electrolyte solution, charged species redistribute themselves to minimize the free energy of the system. In this paper, we explore the Donnan equilibrium of membranes with quaternary electrolyte (Na+/Mg2+/K+/Ca2+/Cl-) solutions, experimentally. The data was used to calculate the ion activity coefficients for six commercial cation-exchange membranes (CEMs). After setting one of the activity coefficients to an arbitrary value, we used the remaining (N-1) activity coefficients as fitting parameters to describe the equilibrium concentrations of (N) ionic species with a mean relative error of 3 %. At increasing solution ionic strengths, the equivalent ion fractions of monovalent counter-ions inside the membrane increased at the expense of the multivalent ones in alignment with the Donnan equilibrium theory. The fitted activity coefficients were employed in a transport model that simulated a Donnan dialysis experiment involving all four cations simultaneously. The arbitrary value assigned to one activity coefficient affects the calculated Donnan potential at the membrane interface. Nevertheless, this arbitrary value does not affect the prediction of the ion concentrations inside the membrane and consequently does not affect the modeled ion fluxes.

Biostimulation with oxygen and electron donors supports micropollutant biodegradation in an experimentally simulated nitrate-reducing aquifer.

The availability of suitable electron donors and acceptors limits micropollutant natural attenuation in oligotrophic groundwater. This study investigated how electron donors with different biodegradability (humics, dextran, acetate, and ammonium), and different oxygen concentrations affect the biodegradation of 15 micropollutants (initial concentration of each micropollutant = 50 µg/L) in simulated nitrate reducing aquifers. Tests mimicking nitrate reducing field conditions showed no micropollutant biodegradation, even with electron donor amendment. However, 2,4-dichlorophenoxyacetic acid and mecoprop were biodegraded under (micro)aerobic conditions with and without electron donor addition. The highest 2,4-dichlorophenoxyacetic acid and mecoprop biodegradation rates and removal efficiencies were obtained under fully aerobic conditions with amendment of an easily biodegradable electron donor. Under microaerobic conditions, however, amendment with easily biodegradable dissolved organic carbon (DOC) inhibited micropollutant biodegradation due to competition between micropollutants and DOC for the limited oxygen available. Microbial community composition was dictated by electron acceptor availability and electron donor amendment, not by micropollutant biodegradation. Low microbial community richness and diversity led to the absence of biodegradation of the other 13 micropollutants (such as bentazon, chloridazon, and carbamazepine). Finally, adaptation and potential growth of biofilms interactively determined the location of the micropollutant removal zone relative to the point of amendment. This study provides new insight on how to stimulate in situ micropollutant biodegradation to remediate oligotrophic groundwaters as well as possible limitations of this process.

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.

Stimulation of hexachlorocyclohexane (HCH) biodegradation in a full scale in situ bioscreen.

The feasibility of a bioscreen for the in situ biodegradation of HCH and its intermediates is demonstrated at a contaminated site in The Netherlands, via the discontinuous addition of methanol as electron donor. An infiltration system was installed and operated at the site over a length of 150 m and a depth of 8 m, to create an anaerobic infiltration zone in which HCH is converted. The construction of the infiltration system was combined with the redevelopment of the site. During passage through the bioscreen, the concentration of HCH in the groundwater decreased from 600 µg/L to the detection limit of the individual HCH isomers (0.01 µg/L) after one year of operation. The concentration of the intermediate biodegradation products benzene and chlorobenzene increased and achieved steady state values of respectively 800 and 2700 µg/L. Benzene and chlorobenzene were treated aerobically on site in an existing wastewater treatment plant. By changing the infiltration regime, it is conclusively shown that HCH removal is the result of the biological degradation and stimulated by the addition of methanol as electron donor. To our knowledge, this is the first successful field demonstration of the stimulated transformation of HCH to intermediates in a full scale anaerobic in situ bioscreen, combined with an aerobic on site treatment to harmless end products.