Structure and function of the bacterial communities during rhizoremediation of hexachlorobenzene in constructed wetlands.

Vertical flow constructed wetlands (VF CWs) are considered to be effective for treating organic pollutants. The rhizosphere of macrophytes such as Phragmites sp., Typha sp. serves as an active and dynamic zone for the microbial degradation of organic pollutants. However, it is still not clear how soil bacterial communities respond to macrophytes and pollutants during the process. For this purpose, the seedlings of Phragmites australis and Typha angustifolia were planted respectively in the VF CWs added with HCB at a dose of 2 mg/kg. During 96 days of cultivation, we monitored hexachlorobenzene (HCB) removal efficiency by GC/MS and the structure of the rhizosphere bacterial communities in the different VF CWs by denaturing gradient gel electrophoresis (DGGE), and constructed bacterial clone library based on PCR-amplified 16S rRNA gene. As expected, the rhizosphere bacterial communities also remained insensitive to HCB exposure in the wetland soil. The diversity of these microbes presented two stages, from the varied up and down to equilibrium in the entire experimental period. Molecular analysis revealed that the phylum Firmicutes dominated over the bacterial communities. The genera that increased under HCB stress included the well-known HCB-degrading bacteria (Pseudomonas sp. and Alcaligenes sp.) and other common bacteria found in contaminated soil but with lesser known practical functions (Burkholderia sp., Lysinibacillus fusiformis, and Bacillus cereus). Furthermore, there was a certain variance in the relative abundances of the bacterial phyla and HCB removal efficiency among different VF CW treatments. The degradation of HCB in T. angustifolia microcosms was faster than that in P. australis and unvegetated wetlands, and the highest bacterial diversity and richness was found in the VF CWs comprising T. angustifolia.

Rapid mechanochemical synthesis of VOx/TiO2 as highly active catalyst for HCB removal.

A rapid (1.5h) one-step ball milling (BM) method was developed to synthesize VOx/TiO2 (VTi-BM) and WOx or MoOx doped VOx/TiO2 (VWTi-BM or VMoTi-BM). Catalytic activity on gaseous POPs removal was tested using hexachlorobenzene (HCB) as surrogate. Catalytic performance decreased in the order of VWTi-BM (T50%=230°C)>VMoTi-BM (260°C)>VTi-BM (270°C)>VTi-WI (300°C; VOx/TiO2 synthesized by wetness impregnation method). The intermediates from oxidation of HCB were analyzed by off gas analysis, from which 2,2,4,5-tetrachloro-4-cyclopentene-1,3-dione (TCCD), dichloromaleic anhydride (DCMA) and tetrachloro-1,4-bezoquinone (TCBQ) were identified. Furthermore, a possible mechanism for the oxidation of HCB over VOx/TiO2 catalysts was proposed. Mechanism studies showed that BM samples possess better dispersion of the VOx species and more surface chemisorbed oxygen. Doping WOx or MoOx into VOx/TiO2 by ball milling can further enhance catalytic performance by increasing surface acid sites.

Removal of PCBs and HCB from contaminated solids using a novel successive self-propagated sintering process.

Thermal treatments are the primary technologies used to remove persistent organic pollutants from contaminated solids. The high energy consumption during continuous heating, required cost for treating the exhaust gas, and potential formation of secondary pollutants during combustion have prevented their implementation. A novel successive self-propagated sintering process was proposed for removing polychlorinated biphenyls (PCBs) and hexachlorobenzene (HCB) from contaminated solids in a low-cost and environmentally friendly way. Nine laboratory-scale experiments involving different initial concentrations of pollutants and solid compositions were performed. Almost all PCBs (>99%) and HCB (>97%) were removed from solids under constant experimental conditions. Varying initial concentrations of PCBs and HCB in the contaminated solids did not influence the removal efficiency of the pollutants; however, the degradation efficiency of pollutants increased as their initial concentrations increased. Although varying levels of PCDD/Fs were detected in the effluent gas, they were all within the emission standard limit.

Simultaneous degradation of toxic refractory organic pesticide and bioelectricity generation using a soil microbial fuel cell.

In this study, the soil microbial fuel cells (MFCs) were constructed in the topsoil contaminated with toxic refractory organic pesticide, hexachlorobenzene (HCB). The performance of electricity generation and HCB degradation in the soil-MFCs were investigated. The HCB degradation pathway was analyzed based on the determination of degradation products and intermediates. Experimental results showed that the HCB removal efficiencies in the three groups (soil MFCs group, open circuit control group and no adding anaerobic sludge blank group) were 71.15%, 52.49% and 38.92%, respectively. The highest detected power density was 77.5 mW/m(2) at the external resistance of 1000 Ω. HCB was degraded via the reductive dechlorination pathway in the soil MFC under anaerobic condition. The existence of the anode promoted electrogenic bacteria to provide more electrons to increase the metabolic reactions rates of anaerobic bacteria was the main way which could promote the removal efficiencies of HCB in soil MFC.

Ball milling synthesized MnOx as highly active catalyst for gaseous POPs removal: significance of mechanochemically induced oxygen vacancies.

A rapid (1.5 h) one-step ball milling (BM) method was developed not only to modify commercial MnO2 via top-down approaches (BM0), but also to bottom-up synthesize MnO(x) by cogrinding of KMnO4 and MnC4H6O4 (BM1) or KMnO4 and MnSO4 (BM2). Catalysts activity on gaseous POPs removal was tested using hexachlorobenzene (HCBz) as surrogate. Catalytic performance decreases in the order of BM2 ≈ BM1 (T90% = 180-200 °C) > BM0 (260 °C) > CMO ≈ cryptomelane MnO2 (>300 °C). Both adsorption and destruction contribute to HCBz removal at 180 °C while destruction prevails at 200-300 °C. Mechanism studies show that destruction activity is lineally correlated with the amount of surface reactive oxygen species (Oads); stability is determined by the removal of surface chloride, which is associated with the mobility of bulk lattice oxygen (Olat); adsorption capacities are linearly correlated with surface area and pore structure. With the aid of extensive characterizations the excellent performance of BM prepared samples can be explained as (1) abundant surface vacancies enhance the generation of Oads; (2) massive bulk vacancies promote the mobility of bulk Olat; (3) large surface area and uniform pore size distribution facilitate the physisorption of HCBz.

Full method validation for the determination of hexachlorobenzene and hexachlorobutadiene in fish tissue by GC-IDMS.

This paper summarizes the validation strategy and the results obtained for the simultaneous determination of hexachlorobenzene (HCB) and hexachlorobutadiene (HCBD) in fish tissue with a maximum of about 10% m/m fat content using a GC-IDMS technique. The method is applicable for the determination of HCB and HCBD at trace levels in different kinds of fish tissue samples in accordance with the requirements of the EU Directive 2008/105/EC establishing Environmental Quality Standard (EQS) levels for biota in aquatic ecosystems (10 ng/g for HCB and 55 ng/g for HCBD). The method validation aimed to assess performance parameters such as linearity, limit of detection/limit of quantification (LOD/LOQ), trueness, selectivity, intermediate precision, repeatability, stability of the extracts and robustness. The validation experiments have been performed by using uncontaminated fish tissue. Trueness was evaluated by using a certified reference material (NIST SRM 1947) (where applicable) and by the standard addition method. Very good linear signal-concentration curves were obtained for both analytes over the whole range of calibration. The repeatability and the intermediate precision of the method, expressed as relative standard deviation (RSD) and calculated at the EQS level, were estimated to be below 3% both for HCB and HCBD. The limits of quantification were 3.7 ng/g for HCB and 15.7 ng/g for HCBD in the fish. An uncertainty budget for the measurement of both HCB and HCBD in fish at about the EQS levels, applying the described method, has been established in the order of 10%. The analytical method and its performance characteristics take into account the requirements of EU Directive 2009/90/EC regarding the establishment of minimum performance criteria for the methods of analysis to be used in the water monitoring activity of the Water Framework Directive. Finally, the validated method was successfully tested on contaminated Silurus glanis from Ebro River (Spain). The method will be used in the homogeneity, stability and interlaboratory comparison studies for the characterization of a new candidate certified reference material.

[Effect of composting organic fertilizer supplies on hexachlorobenzene dechlorination in paddy soils].

A rice pot experiment was conducted in two soils, Hydragric Acrisols (Ac) and Gleyi-Stagnic Anthrosols (An). Three treatments including control and additions of 1% or 2% composting organic fertilizer were designed for each soil. The objective of this research was to evaluate the reductive dechlorination of hexachlorobenzene (HCB) as affected by organic fertilizer supplies in planted paddy soils, and to analyze the relationship between methane production and HCB dechlorination. The results showed that the HCB residues were decreased by 28.6%-30.1% of the initial amounts in Ac, and 47.3% -61.0% in An after 18 weeks of experiment. The amount of HCB and its metabolite uptake by rice plants was only a few thousandths of the initial HCB amount in soils. The main product of HCB dechlorination was pentachlorobenzene (PeCB). The rates of HCB dechlorination in An were higher than those in Ac, which was mainly attributed to the higher pH and dissolved organic carbon (DOC) content of An. The applications of both 1% and 2% composting organic fertilizer showed significant inhibition on PeCB production after the 6th and 10th week in Ac and An, respectively. In both tested soils, no significant difference of PeCB production rates was observed between the applications of 1% and 2% composting organic fertilizer. The role of methanogenic bacteria in HCB dechlorination was condition-dependent.

Gas chromatography triple quadrupole mass spectrometry method for monitoring multiclass organic pollutants in Spanish sewage treatment plants effluents.

In Spain, although more than 50% of urban wastewaters are currently being treated, only half of them are subjected to biological treatments and only 3% undergo advanced treatment technologies. Consequently, the application of more exhaustive wastewater treatment protocols, including the use of new and improved technologies, the application of wider and integrated quality control and water reuse strategies are a priority. We have used as a reference, the European Water Framework Directive (WFD; Directive 2000/60/CE), which establishes a framework for Community action in the field of water policy, setting a list of priority compounds to be monitored in water in order to evaluate their levels. The aim of the present study is to develop and validate a multi-residue method for the analysis of 57 multi-class organic contaminants in wastewater samples using gas chromatography coupled to triple quadrupole mass spectrometry and apply it to evaluate the presence of such compounds in different wastewater treatment plants. The proposed method is based on a sample treatment using liquid-liquid extraction with n-hexane followed by identification, confirmation and quantitation with gas chromatography tandem mass spectrometry using a triple quadrupole analyzer operating in the selected reaction monitoring mode. Three MS/MS transitions were selected for unambiguous confirmation of the target chemicals. The method was validated at two different concentration levels (15 and 150 ng L(-1)) obtaining recovery rates in the range 70-110% in most cases. The limits of quantitation obtained for most of the compounds tested were in the low nanogram per liter range (below 3 ng L(-1) in all cases). Treated and untreated effluent wastewater samples of different origin (industrial, coastal and urban) provided by several sewage treatment plants (STPs) located throughout Spain were tested. Results so far showed that most of the samples assayed did not contain large amount of these contaminants. Hexachlorobenzene was found to be the more frequently detected contaminant in the studied samples, although at levels below 5 µg L(-1).

[Dechlorination of HCB by bimetals based on zero valent iron].

Based on the reducing capacity of zero valent iron, the study investigated the behavior of dechlorination of hexachlorobenzene by bimetals synthetized using Fe with Ag, Pb or Cu as catalysts, respectively. The results showed that bimetals could dechlorinate HCB faster than Fe(0) did, the optimal ratios of Ag/Fe, Pb/Fe and Cu/Fe were 0.2%, 0.5% and 1%. After reacting 2 hours, the dechlorination rates of HCB by Ag/Fe, Pb/Fe and Cu/Fe were 93.5%, 88.5% and 49.6% respectively. The catalyst metal distribution had a great effect on the reductive dechlorination capacity of the bimetal systems, due to more galvanic cells produced by well-distributed catalyst metal and iron. Increasing the amount of bimetal was an effective way to promote HCB dechlorination rate, 88.6% HCB was degraded in 2 h by 0.8 g Pb/Fe while only 38.3% HCB was degraded by 0.1 g Pb/Fe. Besides, HCB dechlorination could be enhanced a little with increasing ionic strength, the HCB dechlorination rates were 93.5%, 98.0% and 98.9% respectively with Na2SO4 concentration at 0, 0.05 and 0.5 mol x L(-1).

Partitioning of hexachlorobenzene in a kaolin/humic acid/surfactant/water system: combined effect of surfactant and soil organic matter.

Understanding the combined effect of soil organic matter (SOM) and surfactants on the partitioning of hydrophobic organic compounds in soil/water systems is important to predict the effectiveness of surfactant-enhanced remediation (SER). In the present study we investigate the partitioning of hexachlorobenzene (HCB) within a humic acid (HA)-coated kaolin/Triton X-100 (TX100)/water system, with special emphasis on the interaction between TX100 and HA, and their combined effect on HCB sorption. HA firstly enhanced then suppressed TX100 sorption to kaolin as the amounts of HA increased, while the addition of TX100 led to a consistent reduction in HA sorption. In the HA-coated kaolin/TX100/water system, TX100 played a primary role in enhancing desorption of HCB, while the role could be suppressed and then enhanced as HA coating amounts increased. Only at HA coating above 2.4%, dissolved HA outcompeted clay-bound HA for HCB partitioning, resulting in dissolved HA enhanced desorption. The presence of dissolved HA at these conditions further promoted the effectiveness of TX100 enhanced desorption. Despite a reduced TX100 sorption to clay was achieved due to the presence of dissolved HA, the effect on HCB desorption was comparatively slight. A reliable cumulative influence of HA and TX100 on HCB desorption was observed, although HCB desorption by HA/TX100 mixed was less than the sum of HA and TX100 individually. Our study suggests that for soils of high organic contents, the combined effect of SOM and surfactants on HOCs desorption can be applied to improve the performance of SER.

Development of self-assembled 3D Fe(x)Oy micro/nano materials for application in hexachlorobenzene degradation.

Self-assembled three dimensional (3D) hierarchical, flower-like iron oxide micro/nano materials, including pure Fe3O4, alpha-Fe2O3/Fe3O4 composite and pure alpha-Fe2O3, were developed to degrade hexachlorobenzene (HCB) at 300 degrees C. It was found that pure Fe3O4 exhibited the highest activity with the reaction rate constant at 0.959 min(-1). Identified degradation products with lower chlorinated benzenes suggest a stepwise hydrodechlorination was occurring for the degradation of HCB on the synthesized Fe3O4. The characterization of the synthesized iron oxides after reaction indicates the Fe3O4 phase was partially transformed into alpha-Fe2O3 phase and in return accelerated progress of hydrodechlorination of HCB. The superior performance of as-prepared Fe3O4 could be attributed to its structural feature such as surface area and pore structure, as well as its phase transformation during HCB degradation.

Remediation of hexachlorobenzene contaminated soils by rhamnolipid enhanced soil washing coupled with activated carbon selective adsorption.

The present study investigates the selective adsorption of hexachlorobenzene (HCB) from rhamnolipid solution by a powdered activated carbon (PAC). A combined soil washing-PAC adsorption technique is further evaluated on the removal of HCB from two soils, a spiked kaolin and a contaminated real soil. PAC at a dosage of 10 g L(-1) could achieve a HCB removal of 80-99% with initial HCB and rhamnolipid concentrations of 1 mg L(-1) and 3.3-25 g L(-1), respectively. The corresponding adsorptive loss of rhamnolipid was 8-19%. Successive soil washing-PAC adsorption tests (new soil sample was subjected to washing for each cycle) showed encouraging leaching and adsorption performances for HCB. When 25 g L(-1) rhamnolipid solution was applied, HCB leaching from soils was 55-71% for three cycles of washing, and HCB removal by PAC was nearly 90%. An overall 86% and 88% removal of HCB were obtained for kaolin and real soil, respectively, by using the combined process to wash one soil sample for twice. Our investigation suggests that coupling AC adsorption with biosurfactant-enhanced soil washing is a promising alternative to remove hydrophobic organic compounds from soils.

Elimination of persistent organic pollutants from fish oil with solid adsorbents.

Fish oils are one of the main sources of ω-3 fatty acids in animal and human diet. However, they can contain high concentrations of persistent organic pollutants due to their lipophilic properties. The aim of this study is the reduction of persistent organic pollutants in fish oil using silicon-based and carbon-based solid adsorbents. A wide screening study with different commercially available adsorbents was carried out, in order to determine their capacity of pollutant removal from fish oil. Moreover, adsorption conditions were evaluated and optimized with using an experimental design and adjustment of the experimental results to response surfaces, obtaining removals rates of more than 99% of PCDD/Fs, 81% of dioxin-like PCBs, 70% of HCB, 41% of DDTs, 16% of marker PCBs and 10% of PBDEs. Finally, fish oil fatty acids were analyzed before and after the treatment with solid adsorbents, confirming that it did not affect its nutritive properties.

Highly purified hexachlorobenzene induces cytochrome P4501A in primary cultures of chicken embryo hepatocytes.

Some uncertainty exists regarding the purity of hexachlorobenzene (HCB) used in past toxicity studies. It has been suggested that reported toxic and biochemical effects initially attributed to HCB exposure may have actually been elicited by contamination of HCB by polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs). Herein, primary cultures of chicken embryo hepatocytes (CEH) were used to compare the potencies of two lots of reagent-grade hexachlorobenzene (HCB-old [HCB-O] and HCB-new [HCB-N]), highly purified HCB (HCB-P) and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) as inducers of ethoxyresorufin O-deethylase (EROD) activity, cytochrome P4501A4 (CYP1A4) messenger ribonucleic acid (mRNA) and CYP1A5 mRNA. The study also compared the EROD- and CYP1A4/5 mRNA-inducing potencies of HCB to the potencies of two mono-ortho substituted polychlorinated biphenyls (PCBs), 2,3,3',4,4'-pentachlorobiphenyl (PCB 105) and 2,3'4,4',5-pentachlorobiphenyl (PCB 118). HCB-O, HCB-N and HCB-P all induced EROD activity and up-regulated CYP1A4 and CYP1A5 mRNAs. Induction was not caused by contamination of HCB with PCDDs or PCDFs. Based upon a comparison of the EC(50) and EC(threshold) values for EROD and CYP1A4/5 mRNA concentration-response curves, the potency of HCB relative to the potency of TCDD was 0.0001, and was similar to that of PCB 105 and PCB 118. The maximal EROD activity and CYP1A4/5 mRNA expression differed greatly between HCB and TCDD, and may contribute to an overestimation of the ReP value calculated for highly purified HCB.

Short-term exposure testing of six different passive samplers for the monitoring of hydrophobic contaminants in water.

Passive sampling devices are increasingly relied upon for monitoring non-polar organic contaminants in water. While many types of devices are available they have seldom been evaluated alongside each other. We tested six passive sampling devices namely: Chemcatcher, two modified versions of the membrane enclosed sorptive coating (MESCO I (m) and MESCO II), silicone rod and strip and semipermeable membrane device (SPMD). Samplers spiked with a range of performance reference compounds (PRCs) were exposed (5 days) in a continuous flow-through tank using Meuse river water fortified with fluctuating concentrations (20-700 ng L(-1)) of polycyclic aromatic hydrocarbons, polychlorinated biphenyls, hexachlorobenzene and p,p'-DDE. Dissipation rates of PRCs appeared to provide reliable information on exchange kinetics even under these short-term exposure conditions. They accounted for differences between masses of contaminants accumulated by replicate samplers, indicating that the variability between replicates was in part due to differences in water turbulences and hence boundary layer thickness. In this system, resistances in the membrane and boundary layers are likely to be in the same order of magnitude for PRCs. Sampler performance was evaluated by comparing masses accumulated in the devices only for analytes for which uptake was linear (integrative) and limited by transport across the boundary layer. Consistent data were obtained across the range of samplers despite their different configurations, and the analysis being conducted in three separate laboratories. The pattern in analyte masses accumulated by Chemcatcher and MESCO II data could be explained by the extraction and analysis being conducted only on the receiving phase of the samplers and a significant impact of the lag-phase prior to obtaining a steady flux of contaminants across the polyethylene membranes.

Pilot-scale electrokinetic movement of HCB and Zn in real contaminated sediments enhanced with hydroxypropyl-beta-cyclodextrin.

This study deals with the efficiency of a pilot-scale electrokinetic (EK) treatment on real aged sediments contaminated with hexachlorobenzene (HCB) and Zn. A total of 0.5m(3) of sediments were treated under a constant voltage in a polyvinyl chloride reactor. The changes of sediment pH, electrical conductivity (EC), organic content (OC), the transport of contaminants in sediments and the consumption of electric energy were evaluated. After 100 d processing, sediment pH slightly increased compared with the initial values, particularly in the bottom layer close to cathodic section, while sediment EC in most sections significantly decreased. Sediment OC in all sections increased, which implied that hydroxypropyl-beta-cyclodextrin (HPCD) was successfully penetrated across sediments by electroosmosis. Significant movement of contaminants was observed across sediments with negligible removals. Both HCB and Zn generally moved from sections near anode and accumulated near cathode. Upon the completion of treatment, the electric energy consumption was calculated as 563 kWhm(-3). This pilot-scale EK test indicates that it is difficult to achieve great removal of hydrophobic organic compounds (HOCs), or HOCs and heavy metal mixed contaminants, by EK treatment in large scale with the use of HPCD.

Hexachlorobenzene removal from a model sediment by using ultrasonic irradiation.

Persistent organic pollutants (POPs) are very difficult to degrade when present in the environment and finally accumulate in the human body. Therefore, it is essential to develop efficient technologies for removal of POPs from polluted sediments. In this study, the authors attempted to evaluate the potential of hexachlorobenzene (HCB; a POP) removal by using ultrasonic irradiation and its combination with other advanced chemical oxidation processes (AOPs) and surfactant addition. The average removal ratios for ultrasonic irradiation alone and ultrasonic in combination with UV light irradiation were almost identical at 40%, and the efficiency could be improved to about 49% by combination with photocatalysis or surfactant addition. The dual-frequency ultrasonic irradiation showed the highest HCB removal ratio.

Removal of hexachlorobenzene from soil by electrokinetically enhanced chemical oxidation.

This study investigates the feasibility of enhanced electrokinetic Fenton process for the remediation of hexachlorobenzene (HCB) in low permeable soil. Laboratory scale experiments were carried out in two different type of experimental setup to evaluate the influence of electrode positions in the system. Kaolin was artificially contaminated with HCB and treated by electrokinetic Fenton process. beta-Cyclodextrin was used to enhance the solubility of HCB in pore fluid. Results show that the position of electrodes in the system and the way in which Fenton's reagent was added to the system has a significant influence on the treatment efficiency.

Solubility-enhanced electrokinetic movement of hexachlorobenzene in sediments: a comparison of cosolvent and cyclodextrin.

Though solubility-enhanced electrokinetics (EK) has been investigated in remediation of soils contaminated with hydrophobic organic chemicals (HOCs), few comparative studies were performed regarding the effect of varied solubilizing agents on both EK parameters and contaminant removal. In this study, performances of two solubilizing agents, ethanol and methyl-beta-cyclodextrin (MCD), were compared in terms of either EK parameters or enhancement of hexachlorobenzene (HCB) movement in real contaminated sediments. Six bench-scale EK tests were conducted under a voltage gradient of 2V cm(-1) for 14 or 21 d. Results reveal that ethanol had a more negative effect on cumulative electroosmotic flow (EOF) than MCD. Furthermore, the distribution of ethanol in the sediment upon the completion of EK tests was lower than that of MCD. The movement of HCB in sediments was observed to increase with increasing concentrations of ethanol or MCD. Test with 50% ethanol exhibited the highest performance, followed by test with 50 g L(-1) MCD. The different performance of HCB removal for tests with varied solubilizing agents was found to be a combined effect of the distribution of solubilizing agents in sediments, the dissolution of HCB by pore liquid and the quantity of cumulative EOF. Finally, an integrated consideration of both EK parameters and contaminant removal suggests that MCD can perform better than ethanol for a long-term field application.

Remediation of hexachlorobenzene in soil by enhanced electrokinetic Fenton process.

The feasibility of enhanced electrokinetic Fenton process for the remediation of Hexachlorobenzene (HCB) in low permeability soil was investigated. Kaolin was artificially contaminated with HCB and treated by electrokinetic and electrokinetic Fenton processes. beta -Cyclodextrin was used to enhance the solubility of HCB in pore fluid. Three tests were carried out, of which two were electrokinetic experiments to observe the suitability of beta -cyclodextrin as a flushing solution for these processes. The third experiment was the electrokinetic Fenton test using beta -cyclodextrin as an enhancing agent. Results show that the removal efficiency depends on the choice of a suitable flushing solution and physical parameters like pH, electric current and electro-osmotic flow.