Biotransformation of Herbicides in the Presence of Various Electron Acceptors

Abstract

Among the important factors affecting the biotransformation of xenobiotic chemicals upon their release into the environment are the dominant electron acceptor condition present and the presence of other, more readily degraded carbon sources. Here, glass-bead biofilm columns were used to investigate the effects of the presence of three different inorganic electron acceptor conditions (oxygen respiration, nitrate reduction, and sulfate reduction) on the biotransformation of the acetanilide herbicides alachlor and propachlor, and to determine the effects of two exogenous carbon sources (acetate and glucose) on their biotransformation under each of these conditions.

Biotransformation of alachlor and propachlor occurred in the presence of both carbon sources and under each of the three electron acceptor conditions. Both were transformed most rapidly under sulfate-reducing conditions. Analysis by gas chromatography/mass spectrometry (GC/MS) did not reveal any significant metabolic products. Both herbicides react abiotically with bisulfide, produced within the sulfate-reducing cultures, though most of the transformation was attributed to the microorganisms. The primary, readily degraded carbon source (acetate or glucose) was needed to establish each culture, and its continuous presence was required to sustain herbicide biotransformation in the sulfate-reducing reactors. Loss of either acetate or glucose from the column influent did not significantly affect herbicide biotransformation in the aerobic or nitratereducing reactors, at least for short periods. Temporary loss of the external electron acceptors (O₂, NO^- ₃or SO₄ ^2-) slowed transformation of both herbicides.     

金属铁还原脱氯处理有机氯化物的研究进展

本文对金属铁在还原降解六氯乙烷、四氯化碳、氯仿、三氯乙烯、四氯乙烯、二氯乙烯、氯乙烯、五氯酚、多氯联苯等有机氯化物中的应用作了评述 ,讨论了金属铁降解有机氯化物的反应机理及动力学。     

Biodegradation of Kupferschiefer black shale organic matter (Fore-Sudetic Monocline, Poland) by indigenous microorganisms

This study provides the first evidence for the direct biodegradation of persistent organic matter extracted from the organic-rich polymetallic black shale ore Kupferschiefer, one of the most important sources of metals in the world. It was demonstrated that an enriched community of indigenous heterotrophic microorganisms isolated from black shale grown under aerobic conditions could utilize shale organic matter as the sole carbon and energy source. Colonization of shale organic matter was observed. The main biodegradation intermediates and products such as phosphonic acid dioctadecyl ester and isoindole-1,3 were detected in the aqueous phase of cultures. The bacterial community showed the ability to PAH biodegradation, assimilation of organic acids and esters as well as lipase activity. The intracellular accumulation of phosphorus by bacteria during growth on organic matter was confirmed. Strains within the genus Pseudomonas were found to dominate the bacterial population at the end of the experiment. The results of this study confirm that indigenous bacteria are likely to play a role in the biotransformation of black shale and can influence the geochemical cycles of ancient organic carbon in the deep terrestrial subsurface. This process may also occur in tailings ponds containing black shale, and cause the mobilization of potentially toxic compounds to the soil and groundwater.     

Anaerobic decomposition of halogenated aromatic compounds

Halogenated compounds constitute one of the largest groups of environmental pollutants, partly as a result of their widespread use as biocides, solvents and other industrial chemicals. A critical step in degradation of organohalides is the cleavage of the carbon?halogen bond. Reductive dehalogenation is generally the initial step in metabolism under methanogenic conditions, which requires a source of reducing equivalents, with the halogenated compound serving as an electron acceptor. Dehalogenation is greatly influenced by alternate electron acceptors; e.g. sulfate frequently inhibits reductive dehalogenation. On the other hand, a number of halogenated aromatic compounds can be degraded under different electron-accepting conditions and their complete oxidation to CO(2) can be coupled to processes such as denitrification, iron(III)-reduction, sulfate reduction and methanogenesis. Reductive dehalogenation was the initial step in degradation not only under methanogenic, but also under sulfate- and iron(III)-reducing conditions. Dehalogenation rates were in general slower under sulfidogenic and iron-reducing conditions, suggesting that dehalogenation was affected by the electron acceptor. The capacity for dehalogenation appears to be widely distributed in anoxic environments; however, the different substrate specificities and activities observed for the halogenated aromatic compounds suggest that distinct dehalogenating microbial populations are enriched under the different reducing conditions. Characterization of the microbial community structure using a combination of biomolecular techniques, such as cellular fatty acid profiling, and 16 S rRNA fingerprinting/sequence analysis, was used to discern the distinct populations enriched with each substrate and under each electron-accepting condition. These combined techniques will aid in identifying the organisms responsible for dehalogenation and degradation of halogenated aromatic compounds.     

固态碳源去除地下水硝酸盐的模拟实验

选取了5种研究较少的固体材料,棉花、丝瓜络、甘蔗渣、可降解餐盒、木屑作为去除地下水硝酸盐的外加碳源。在锥形瓶中进行反硝化对比实验,研究了不同固态碳源下NO3--N、NO2--N、NH4+-N及pH的变化情况,分析了NO3--N及总氮的去除率。研究结果表明,反硝化过程中pH呈升高趋势,在6.9～8.5范围内浮动。可降解餐盒和丝瓜络相对于其他的固态碳源来说,对NO3--N和总氮有较高的去除率,但丝瓜络的总氮去除率明显低于可降解餐盒。可降解餐盒的硝酸盐去除率达到98.28%,总氮去除率达到93.48%。可降解餐盒能够有效地去除地下水硝酸盐,达到以废治废的效果,是经济有效的最佳固态碳源。     

Formation of chloroform and carbon tetrachloride in foods treated with sodium hypochlorite

The formation of chloroform and carbon tetrachloride during the treatment of fresh vegetable homogenate with sodium hypochlorite was studied.When a n-hexane extract of the reaction mixture of fresh vegetables and sodium hypochlorite was analysed by gas chromatography, chloroform and carbon tetrachloride were detected, and many unidentified peaks were also observed. The formation conditions of chloroform and carbon tetrachloride were studied. The mechanism of chloroform formation may be different from that of carbon tetrachloride according to the study of pH on the formation of both compounds. The formation of chloroform is much greater than that of carbon tetrachoride.     

Assessing the transformation of chlorinated ethenes in aquifers with limited potential for natural attenuation: added values of compound-specific carbon isotope analysis and groundwater dating

The evaluation of biotransformation of chlorinated ethenes (CEs) in contaminated aquifers is challenging when variable redox conditions and groundwater flow regime are limiting factors. By using compound-specific stable carbon isotope analysis (C-CSIA) and ³H-³He based groundwater dating, we assessed three CE-contaminated field sites that differed in groundwater flow velocities, redox conditions, and level of contamination. CE isotopic signatures and carbon isotopic mass balances were applied to quantify CE transformation, whereas groundwater dating allowed determining degradation timescales and assessing hydrodynamic regimes. The combination of these techniques enabled at all field sites to indicate zones within the aquifers where CE dechlorination preferably occurred, sometimes even to metabolites of no toxic concern. However, the natural transformation processes were insufficient to mitigate the entire CE contamination at the studied sites. Such situations of limited transformation are worldwide far more common than sites where optimal natural (mainly redox) conditions are enabling complete CEs degradation. Despite such constraints for natural transformation, this study showed that even under non-favorable biogeochemical CEs degradation, the combination of CSIA and groundwater dating provide valuable information to the understanding of the fate of the CEs, thus, being an important contribution in the definition of efficient remediation measures at any given biogeochemical conditions.     

Attenuation reactions in a multiple contaminated aquifer in Bitterfeld (Germany)

Large-scale contaminated sites with multiple contaminants in the groundwater present a challenge to risk assessment and remediation. Attenuation reactions take place in the subsurface and act to contain contaminants, but must be thoroughly investigated on a site-specific basis. Field data from monitoring wells at a contaminated industrial site in Bitterfeld, Germany, are presented and analyzed for evidence of the prevalent biodegradation reactions. The groundwater in the Tertiary aquifer is contaminated with large quantities of chlorinated aliphatic compounds, in addition to chlorobenzenes and BTEX. In this strictly anaerobic environment, geochemical indications for several microbial processes were found, including methanogenesis, sulfate and iron reduction as well as reductive dechlorination of the chlorinated hydrocarbons. Direct evidence for the latter degradation reaction was observed along the flowpath due to the appearance of intermediates and an increase in the degree of dechlorination.     

Removal of low concentrations of carbon tetrachloride in compost-based biofilters operated under methanogenic conditions.

Research was performed to demonstrate the removal of carbon tetrachloride (CT) using compost biofilters operated under methanogenic conditions. Biofilters were operated at an empty-bed residence time of 2.8 minutes using nitrogen as the atmosphere. Hydrogen and carbon dioxide were supplied as an electron donor and carbon source, respectively, during acclimation of the bed medium microbes. Once methanogenesis was demonstrated, CT flow to the biofilter was established. Biofilters were operated over a CT concentration range from 20 to 700 ppbv for 6 months. Bed medium microbes were able to remove up to 75% of the inlet CT. At excessively high CT concentrations (> 500 ppmv), methane production and hydrogen utilization by the bed medium microbes appeared to be inhibited. CT removal by the biofilter decreased when the hydrogen supply was removed from the biofilter inlet, indicating that hydrogen acted as the electron donor for reductive dechlorination. The removal efficiency and relatively low empty bed residence times demonstrated by these laboratory-scale biofilters indicate that anaerobic biofiltration of CT may be a feasible full-scale process.     

Removal of Low Concentrations of Carbon Tetrachloride in Compost-Based Biofilters Operated under Methanogenic Conditions

ABSTRACT

Research was performed to demonstrate the removal of carbon tetrachloride (CT) using compost biofilters operated under methanogenic conditions. Biofilters were operated at an empty-bed residence time of 2.8 minutes using nitrogen as the atmosphere. Hydrogen and carbon dioxide were supplied as an electron donor and carbon source, respectively, during acclimation of the bed medium microbes. Once methanogenesis was demonstrated, CT flow to the biofilter was established. Biofilters were operated over a CT concentration range from 20 to 700 ppbv for 6 months. Bed medium microbes were able to remove up to 75% of the inlet CT. At excessively high CT concentrations (>500 ppmv), methane production and hydrogen utilization by the bed medium microbes appeared to be inhibited. CT removal by the biofilter decreased when the hydrogen supply was removed from the biofilter inlet, indicating that hydrogen acted as the electron donor for reductive dechlorination. The removal efficiency and relatively low empty bed residence times demonstrated by these laboratory-scale biofilters indicate that anaerobic biofiltration of CT may be a feasible full-scale process.     

Microbially mediated redox processes in natural analogues for radioactive waste

Natural analogues allow scientists to investigate biogeochemical processes relevant to radioactive waste disposal that occur on time scales longer than those that may be studied by time-limited laboratory experiments. The Palmottu U-Th deposit in Finland and the Bangombé natural nuclear reactor in Gabon involve the study of natural uranium, and are both considered natural analogues for subsurface radioactive waste disposal. The microbial population naturally present in groundwater may affect the redox conditions, and hence, the radionuclide solubility and migration. Therefore, groundwater samples from the two sites were investigated for microbial populations. The total numbers of cells ranged from 10(4) to 10(6) cells ml(-1). Iron-reducing bacteria (IRB) were the largest culturable microbial population in the Palmottu groundwater and were present at up to 1.3 x 10(5) cells ml(-1). Sulfate-reducing bacteria (SRB) and acetogens could also be cultured from the Palmottu groundwater. The numbers of IRB and SRB were largest in groundwater with the lowest uranium concentrations. Removal of dissolved U(VI) from solution was concomitant with the growth of IRB enrichment cultures and the reduction of iron. The redox buffer in the Palmottu groundwater consists of iron and uranium species, both of which are affected by IRB. IRB and aerobic heterotrophs were cultured from the Bangombé groundwater, where redox potentials are buffered by iron and organic carbon species. Microbial populations similar to those found at Palmottu and Bangombé are found throughout the Fennoscandian Shield, a potential host rock for subsurface radioactive waste disposal. These results confirm that microorganisms can be expected to play a role in stabilizing radioactive waste disposed of in the subsurface by lowering redox potential and immobilizing radionuclides.     

Numerical simulation of a fine-grained denitrification layer for removing septic system nitrate from shallow groundwater

One of the most common methods to dispose of domestic wastewater involves the release of septic effluent from drains located in the unsaturated zone. Nitrogen from such systems is currently of concern because of nitrate contamination of drinking water supplies and eutrophication of coastal waters. It has been proposed that adding labile carbon sources to septic distribution fields could enhance heterotrophic denitrification and thus reduce nitrate concentrations in shallow groundwater. In this study, a numerical model which solves for variably saturated flow and reactive transport of multiple species is employed to investigate the performance of a drain field design that incorporates a fine-grained denitrification layer. The hydrogeological scenario simulated is an unconfined sand aquifer. The model results suggest that the denitrification layer, supplemented with labile organic carbon, may be an effective means to eliminate nitrogen loading to shallow groundwater. It is also shown that in noncalcareous aquifers, the denitrification reaction may provide sufficient buffering capacity to maintain near neutral pH conditions beneath and down gradient of the drain field. Leaching of excess dissolved organic carbon (DOC) from the denitrification layer is problematic, and causes an anaerobic plume to develop in simulations where the water table is less than 5-6 m below ground surface; this anaerobic plume may lead to other down gradient changes in groundwater quality. A drain field and denitrification layer of smaller dimensions is shown to be just as effective for reducing nitrate, but has the benefit of reducing the excess DOC leached from the layer. This configuration will minimize the impact of wastewater disposal in areas where the water table is as shallow as 3.5 m.     

电动力学技术强化原位生物修复研究进展

介绍了利用电动力学技术强化土壤及地下水原位生物修复的原理和最新进展。电动力学强化的基本原理是利用电渗析、电迁移和电泳等电动力学效应加速污染环境中有机污染物和微生物运动,注入营养物、电子受体或活性微生物,或者利用电极反应和电流热效应为地下生物降解创造有利条件。研究表明．电动力学技术能有效地强化原位生物修复,而且该技术不破环生态环境．安装和操作简单,成本低廉．有广泛的应用前景。     

Degradation of carbon tetrachloride in the presence of iron and sulphur containing compounds

The effect of several sulphur compounds: sodium sulphate, sodium sulphide, ferrous sulphide,pyrite and an organosulphonic acid on the kinetics of the iron (Fe °) induced degradation of carbon tetrachloride was examined under aerobic conditions. It was observed that all of the sulphur compounds investigated significantly accelerated the reaction. The mechanisms of the processes studied as well as their possible influence on the efficiency of the iron-induced dehalogenation of pollutants, both in situ and in above-ground treatment are discussed.     

Humic colloid-borne migration of uranium in sand columns

Column experiments were carried out to investigate the influence of humic colloids on subsurface uranium migration. The columns were packed with well-characterized aeolian quartz sand and equilibrated with groundwater rich in humic colloids (dissolved organic carbon (DOC): 30 mg dm(-3)). U migration was studied under an Ar/1% CO2 gas atmosphere as a function of the migration time, which was controlled by the flow velocity or the column length. In addition, the contact time of U with groundwater prior to introduction into a column was varied. U(VI) was found to be the dominant oxidation state in the spiked groundwater. The breakthrough curves indicate that U was transported as a humic colloid-borne species with a velocity up to 5% faster than the mean groundwater flow. The fraction of humic colloid-borne species increases with increasing prior contact time and also with decreasing migration time. The migration behavior was attributed to a kinetically controlled association/dissociation of U onto and from humic colloids and also a subsequent sorption of U onto the sediment surface. The column experiments provide an insight into humic colloid-mediated U migration in subsurface aquifers.     

Hydraulic constraints on the performance of a groundwater denitrification wall for nitrate removal from shallow groundwater

Denitrification walls are a practical approach for decreasing non-point source pollution of surface waters. They are constructed by digging a trench perpendicular to groundwater flow and mixing the aquifer material with organic matter, such as sawdust, which acts as a carbon source to stimulate denitrification. For efficient functioning, walls need to be permeable to groundwater flow. We examined the functioning of a denitrification wall constructed in an aquifer consisting of coarse sands. Wells were monitored for changes in nitrate concentration as groundwater passed through the wall and soil samples were taken to measure microbial parameters inside the wall. Nitrate concentrations upstream of the wall ranged from 21 to 39 g N m(-3), in the wall from 0 to 2 g N m(-3) and downstream from 19 to 44 g N m(-3). An initial groundwater flow investigation using a salt tracer dilution technique showed that the flow through the wall was less than 4% of the flow occurring in the aquifer. Natural gradient tracer tests using bromide and Rhodamine-WT confirmed groundwater bypass under the wall. Hydraulic conductivity of 0.48 m day(-1) was measured inside the wall, whereas the surrounding aquifer had a hydraulic conductivity of 65.4 m day(-1). This indicated that during construction of the wall, hydraulic conductivity of the aquifer had been greatly reduced, so that most of the groundwater flowed under rather than through the wall. Denitrification rates measured in the center of the wall ranged from 0.020 to 0.13 g N m(-3) day(-1), which did not account for the rates of nitrate removal (0.16-0.29 g N m(-3) day(-1)) calculated from monitoring of groundwater nitrate concentrations. This suggested that the rate of denitrification was greater at the upstream face of the wall than in its center where it was limited by low nitrate concentrations. While denitrification walls can be an inexpensive tool for removing nitrate from groundwater, they may not be suitable in aquifers with coarse textured subsoils where simple inexpensive construction techniques result in major decreases in hydraulic conductivity.     

Aerobic biodegradation of dichloroethenes by indigenous bacteria isolated from contaminated sites in Africa

The widespread use of tetrachloroethene (PCE) and trichloroethene (TCE) as dry cleaning solvents and degreasing agents for military and industrial applications has resulted in significant environmental contamination worldwide. Anaerobic biotransformation of PCE and TCE through reductive dechlorination frequently lead to the accumulation of dichloroethenes (DCEs), thus limiting the use of reductive dechlorination for the biotransformation of the compounds. In this study, seven bacteria indigenous to contaminated sites in Africa were characterized for DCE degradation under aerobic conditions. The specific growth rate constants of the bacterial isolates ranged between 0.346-0.552d(-1) and 0.461-0.667d(-1) in cis-DCE and trans-DCE, respectively. Gas chromatographic analysis revealed that up to 75% of the compounds were degraded within seven days with the degradation rate constants ranging between 0.167 and 0.198d(-1). The two compounds were also observed to be significantly degraded, simultaneously, rather than sequentially, when present as a mixture. Phylogenetic analysis of the 16S rRNA gene sequences of the bacterial isolates revealed their identity as well as their relation to other environmentally-important bacteria. The observed biodegradation of DCEs may contribute to PCE and TCE removal at the aerobic fringe of groundwater plumes undergoing reductive dechlorination in contaminated sites.     

A strategy for aromatic hydrocarbon bioremediation under anaerobic conditions and the impacts of ethanol: a microcosm study

Increased use of ethanol-blended gasoline (gasohol) and its potential release into the subsurface have spurred interest in studying the biodegradation of and interactions between ethanol and gasoline components such as benzene, toluene, ethylbenzene and xylene isomers (BTEX) in groundwater plumes. The preferred substrate status and the high biological oxygen demand (BOD) posed by ethanol and its biodegradation products suggests that anaerobic electron acceptors (EAs) will be required to support in situ bioremediation of BTEX. To develop a strategy for aromatic hydrocarbon bioremediation and to understand the impacts of ethanol on BTEX biodegradation under strictly anaerobic conditions, a microcosm experiment was conducted using pristine aquifer sand and groundwater obtained from Canadian Forces Base Borden, Canada. The initial electron accepter pool included nitrate, sulfate and/or ferric iron. The microcosms typically contained 400 g of sediment, 600 approximately 800 ml of groundwater, and with differing EAs added, and were run under anaerobic conditions. Ethanol was added to some at concentrations of 500 and 5000 mg/L. Trends for biodegradation of aromatic hydrocarbons for the Borden aquifer material were first developed in the absence of ethanol, The results showed that indigenous microorganisms could degrade all aromatic hydrocarbons (BTEX and trimethylbenzene isomers-TMB) under nitrate- and ferric iron-combined conditions, but not under sulfate-reducing conditions. Toluene, ethylbenzene and m/p-xylene were biodegraded under denitrifying conditions. However, the persistence of benzene indicated that enhancing denitrification alone was insufficient. Both benzene and o-xylene biodegraded significantly under iron-reducing conditions, but only after denitrification had removed other aromatics. For the trimethylbenzene isomers, 1,3,5-TMB biodegradation was found under denitrifying and then iron-reducing conditions. Biodegradation of 1,2,3-TMB or 1,2,4-TMB was slower under iron-reducing conditions. This study suggests that addition of excess ferric iron combined with limited nitrate has promise for in situ bioremediation of BTEX and TMB in the Borden aquifer and possibly for other sites contaminated by hydrocarbons. This study is the first to report 1,2,3-TMB biodegradation under strictly anaerobic condition. With the addition of 500 mg/L ethanol but without EA addition, ethanol and its main intermediate, acetate, were quickly biodegraded within 41 d with methane as a major product. Ethanol initially present at 5000 mg/L without EA addition declined slowly with the persistence of unidentified volatile fatty acids, likely propionate and butyrate, but less methane. In contrast, all ethanol disappeared with repeated additions of either nitrate or ferric iron, but acetate and unidentified intermediates persisted under iron-enhanced conditions. With the addition of 500 mg/L ethanol and nitrate, only minor toluene biodegradation was observed under denitrifying conditions and only after ethanol and acetate were utilized. The higher ethanol concentration (5000 mg/L) essentially shut down BTEX biodegradation likely due to high EA demand provided by ethanol and its intermediates. The negative findings for anaerobic BTEX biodegradation in the presence of ethanol and/or its biodegradation products are in contrast to recent research reported by Da Silva et al. [Da Silva, M.L.B., Ruiz-Aguilar, G.M.L., Alvarez, P.J.J., 2005. Enhanced anaerobic biodegradation of BTEX-ethanol mixtures in aquifer columns amended with sulfate, chelated ferric iron or nitrate. Biodegradation. 16, 105-114]. Our results suggest that the apparent conservation of high residual labile carbon as biodegradation products such as acetate makes natural attenuation of aromatics less effective, and makes subsequent addition of EAs to promote in situ BTEX biodegradation problematic.     

Fate of nitrogen for subsurface drip dispersal of effluent from small wastewater systems

Subsurface drip irrigation systems apply effluent from onsite wastewater systems in a more uniform manner at a lower rate than has been possible with other effluent dispersal methods. The effluent is dispersed in a biologically active part of the soil profile for optimal treatment and where the water and nutrients can be utilized by landscape plants. Container tests were performed to determine the fate of water and nitrogen compounds applied to packed loamy sand, sandy loam, and silt loam soils. Nitrogen removal rates measured in the container tests ranged from 63 to 95% despite relatively low levels of available carbon. A Hydrus 2D vadose zone model with nitrification and denitrification rate coefficients calculated as a function of soil moisture content fit the container test results reasonably well. Model results were sensitive to the denitrification rate moisture content function. Two-phase transport parameters were needed to model the preferential flow conditions in the finer soils. Applying the model to generic soil types, the greatest nitrogen losses (30 to 70%) were predicted for medium to fine texture soils and soils with restrictive layers or capillary breaks. The slow transport with subsurface drip irrigation enhanced total nitrogen losses and plant nitrogen uptake opportunity.     

Microbial biotransformation of aryl sulfanylpentafluorides

We report, for the first time, the biotransformation of potential pollutants bearing the pentafluorosulfanyl (SF₅-) functional group in a fungus and bacteria. Cunninghamella elegans transformed p-methoxy phenyl SF₅ via demethylation; Pseudomonas knackmussii and P. pseudoalcaligenes KF707 transformed amino-, hydroxyamino- and diamino- substituted phenyl SF₅, forming the N-acetylated derivatives as the main product. Cell-free extract of Streptomyces griseus transformed 4-amino-3-hydroxy-phenyl SF₅ to the N-acetylated derivative in the presence of acetyl CoA, confirming that an N-acetyltransferase is responsible for the bacterial biotransformations. Approximately 25 % of drugs and 30 % of agrochemicals contain fluorine, and the trifluoromethyl group is a prominent feature of many of these since it improves lipophilicity and stability. The pentafluorosulfanyl substituent is seen as an improvement on the trifluoromethyl group and research efforts are underway to develop synthetic methods to incorporate this moiety into biologically active compounds. It is important to determine the potential environmental impact of these compounds, including the potential biotransformation reactions that may occur when they are exposed to microorganisms.