Anaerobic biodegradation of biphenyl in various paddy soils and river sediment

The anaerobic degradation of biphenyl was investigated in four uncontaminated Japanese paddy soils and one river sediment sample contaminated with benzene and chlorinated aliphatics. Two of the paddy soils and the sediment were capable of degrading biphenyl anaerobically without any additional medium or electron acceptors. The half-lives of biphenyl biodegradation in the three samples were 212 d in the Kuridashi soil, 327 d in the Kamajima soil, and 429 d in the river sediment. The Kuridashi soil metabolized 1+/-0.3% of [U-14C]-biphenyl into CO2 and 5+/-2% into water-soluble metabolites after 45 d of incubation. Submerged conditions, which result in lower nitrate and iron oxide contents, and neutral pH, appeared to be the common properties among the samples that influenced their degradation capacities. The addition of 10mM sulfate and 20mM Fe(III) as electron acceptors did not enhance the biphenyl degradation rate, whereas 10mM nitrate completely inhibited biphenyl degradation. The addition of different electron donors (lactate, acetate, or pyruvate) slightly slowed the degradation. Molybdate (an inhibitor of sulfate-reducing bacteria) had an inhibitory effect on biphenyl biodegradation, but bromoethanesulfonic acid (an inhibitor of methanogens) did not. Most biphenyl degradation was observed when only water was added, with no other electron acceptors or donors. These results suggest that sulfate-reducing bacteria and fermentative microbial populations play important roles in anaerobic biphenyl biodegradation in paddy soil.     

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.     

Anaerobic PAH degradation in soil by a mixed bacterial consortium under denitrifying conditions

Polycyclic aromatic hydrocarbons (PAHs) are one of the main classes of contaminants in the terrestrial environment. Concentrations of biphenyl, fluorene, phenanthrene and pyrene were added to soil samples in order to investigate the anaerobic degradation potential of PAHs under denitrifying conditions. A mixed population of microorganisms obtained from a paddy soil was incubated for 20 days in anaerobic conditions in the presence of soil alone or with nitrate, adding, as electron donors, PAHs and, in some samples, glucose or acetate. At regular time intervals oxidation-reduction potential, PAHs concentration, microbial ATP and nitrate concentration into the solution were measured. Degradation trends for each hydrocarbon are similar under all conditions, indicating that the molecular conformation prevails over other parameters in controlling the degradation. Poor degradation results were obtained when PAHs were the only organic matter available for the inoculum, thus confirming the recalcitrance to degradation of these compounds. Biodegradation was influenced by the addition of other carbon sources. As better degradation results were generally obtained when acetate or glucose were added, the hypothesis of a co-metabolic enhancement of PAH biodegradation seems likely. Thus, anaerobic biodegradation of PAHs studied, biphenyl, fluorene, phenanthrene and pyrene, seems to be possible both through fermentative and respiratory metabolism, provided that low molecular weight co-metabolites and suitable electron acceptors (nitrate) are present.     

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.     

Degradation of 2-chlorophenol via a hydrogenotrophic biofilm under different reductive conditions

This research studies the 2-chlorophenol (2-CP) degradation by the hydrogenotrophic biofilm cultivated in three silicone-tube membrane bioreactors under the conditions of denitrification (DN), sulfate-reduction (DS) and dechlorination (DC). Experimental results showed that after acclimation for more than four months with 2-CP, the respective 2-CP removal efficiency was 95% in DN, 94% in DS and 95% in DC reactors, under the condition of influent 2-CP 25 mg/l with hydraulic retention time (HRT) of 15 h. The metabolic pathway of 2-CP was different in different reactors. The 2-CP was thought to be utilized as carbon and energy source in DN and DS reactors, while the dechlorination occurred in the DC reactor in lack of nitrate and sulfate. The pH dramatically affected the 2-CP degradation in all reactors. Experimental results showed that the optimal pH range was around 6+/-0.2 in DN, 7+/-0.2 in DS, and 5.8-7.2 in DC reactors. Both nitrate and sulfate inhibited the 2-CP dechlorination, but the inhibition levels were different. Nitrate completely inhibited the dechlorination at once, while sulfate took a longer time to reach complete inhibition, only after the bacteria were adapted to the sulfate-reducing condition. Both inhibitions were accomplished by taking the place of 2-CP as electron acceptors. H2 served as an electron donor for dechlorination of 2-CP. The dechlorination was apparently stopped when lacking H2 and another pathway was responsible for the 2-CP degradation.     

Hydrochemical and isotopic effects associated with petroleum fuel biodegradation pathways in a chalk aquifer

Hydrochemical data, compound specific carbon isotope analysis and isotopic enrichment trends in dissolved hydrocarbons and residual electron acceptors have been used to deduce BTEX and MTBE degradation pathways in a fractured chalk aquifer. BTEX compounds are mineralised sequentially within specific redox environments, with changes in electron acceptor utilisation being defined by the exhaustion of specific BTEX components. A zone of oxygen and nitrate exhaustion extends approximately 100 m downstream from the plume source, with residual sulphate, toluene, ethylbenzene and xylene. Within this zone complete removal of the TEX components occurs by bacterial sulphate reduction, with sulphur and oxygen isotopic enrichment of residual sulphate (epsilon(s) = -14.4 per thousand to -16.0 per thousand). Towards the plume margins and at greater distance along the plume flow path nitrate concentrations increase with delta15N values of up to +40 per thousand indicating extensive denitrification. Benzene and MTBE persist into the denitrification zone, with carbon isotope enrichment of benzene indicating biodegradation along the flow path. A Rayleigh kinetic isotope enrichment model for 13C-enrichment of residual benzene gives an apparent epsilon value of -0.66 per thousand. MTBE shows no significant isotopic enrichment (delta13C = -29.3 per thousand to -30.7 per thousand) and is isotopically similar to a refinery sample (delta13C = -30.1 per thousand). No significant isotopic variation in dissolved MTBE implies that either the magnitude of any biodegradation-induced isotopic fractionation is small, or that relatively little degradation has taken place in the presence of BTEX hydrocarbons. It is possible, however, that MTBE degradation occurs under aerobic conditions in the absence of BTEX since no groundwater samples were taken with co-existing MTBE and oxygen. Low benzene delta13C values are correlated with high sulphate delta34S, indicating that little benzene degradation has occurred in the sulphate reduction zone. Benzene degradation may be associated with denitrification since increased benzene delta13C is associated with increased delta15N in residual nitrate. Re-supply of electron acceptors by diffusion from the matrix into fractures and dispersive mixing is an important constraint on degradation rates and natural attenuation capacity in this dual-porosity aquifer.     

Denitrification using PBS as carbon source and biofilm support in a packed-bed bioreactor

Biodegradable polymer was used as carbon source and biofilm support for nitrate removal from aqueous solution as an attractive alternative for biological denitrification. The objective of this paper was to investigate the denitrification performance and microbial community of a packed-bed bioreactor using poly (butanediol succinate) (PBS), a biodegradable polymer, as carbon source and biofilm support. NO₃–N concentration was determined by UV spectrophotometer. NO₂–N concentration was assayed by hydrochloric acid naphthyl ethylenediamine spectrophotometry method. Total organic carbon (TOC) was measured using a TOC analyzer. The morphology of the samples was observed using an environmental scanning electron microscope (ESEM). The microbial community was analyzed by pyrosequencing method. The experimental results showed that an average removal efficiency of nitrate was 95 %. ESEM observation and FTIR analysis indicated the changes of PBS granules before and after microbial utilization. Pyrosequencing results showed that Betaproteobacteria predominated, and most of PBS-degrading denitrifying bacteria were assigned to the family Comamonadaceae. Denitrifying bacteria accounted for 13.02 % in total population. The PBS granules were suitable support and carbon source for denitrifying bacteria.     

Nitrate removal from groundwater using biodegradable polymers as carbon source and biofilm support

A new kind of biodegradable polymer, PBS, was used as both carbon source and biofilm support in a fixed-bed reactor to remove nitrate from groundwater. The experimental results showed that the denitrifying bacteria can easily attach to the surface of PBS granules and adapt to use PBS as carbon source. The SEM observation indicated that the fine biofilm can develop on the surface of PBS granules within 15 days. The denitrification rate increased with temperature increase in the range of 10-35°C, the maximum denitrification rate reached 1.00 mg/g.d at 35°C. Continuous experiment results showed that nitrate (50 mg/l) can be removed effectively and nitrite did not accumulate in the effluent.     

Role of cosubstrate and bioaccessibility played in the enhanced anaerobic biodegradation of organochlorine pesticides (OCPs) in a paddy soil by nitrate and methyl-β-cyclodextrin amendments

The present study was conducted to investigate the anaerobic biodegradation potential of biostimulation by nitrate (KNO₃) and methyl-β-cyclodextrin (MCD) addition on an aged organochlorine pesticide (OCP)-contaminated paddy soil. After 180 days of incubation, total OCP biodegradation was highest in soil receiving the addition of nitrate and MCD simultaneously and then followed by nitrate addition, MCD addition, and control. The highest biodegradation of chlordanes, hexachlorocyclohexanes, endosulfans, and total OCPs was 74.3, 63.5, 51.2, and 65.1 %, respectively. Meanwhile, MCD addition significantly increased OCP bioaccessibility (p?<?0.05) evaluated by Tenax TA extraction and a three-compartment model method. Moreover, the addition of nitrate and MCD also obtained the highest values of soil microbial activities, including soil microbial biomass carbon and nitrogen, ATP production, denitrifying bacteria count, and nitrate reductase activity. Such similar trend between OCP biodegradation and soil-denitrifying activities suggests a close relationship between OCP biodegradation and N cycling and the indirect/direct involvement of soil microorganisms, especially denitrifying microorganisms in the anaerobic biodegradation of OCPs.     

Anaerobic degradation of pyrrolidine and piperidine coupled with nitrate reduction

Biodegradability of secondary amines (pyrrolidine, piperidine, piperazine, morpholine, and thiomorpholine) under anaerobic conditions was examined in microbial consortia from six different environmental sites. The consortia degraded pyrrolidine and piperidine under denitrifying conditions. Enrichment cultures were established by repeatedly sub-culturing the consortia on pyrrolidine or piperidine in the presence of nitrate. The enrichments strictly required nitrate for the anaerobic degradation and utilized pyrrolidine or piperidine as a carbon, nitrogen, and energy source for their anaerobic growths. The anaerobic degradation of pyrrolidine and piperidine reduced nitrate to nitrogen gas, indicating that these anaerobic degradations were coupled with a respiratory nitrate reduction.     

A method for measuring the anoxic biodegradability under denitrifying conditions

A test for assessing the anoxic biodegradability of organic compounds under denitrifying conditions is proposed. The method is based on the recovery and quantification of the CO2 produced, which is evidence of complete biodegradation of the test compound (added as the sole carbon source). The tests were carried out in a mineral medium, with nitrate as electron acceptor. Whole lake sediments, sediment extracts and a commercial inoculum were assayed as a possible inoculum source by means of glucose biodegradability tests. It was found that the sediment extracts constitute a suitable and environmentally-relevant inoculum source, since they add non-significant amounts of carbon to the tests. Two xenobiotic compounds, namely, aniline and phenol, were tested in the aforementioned conditions as well as in a standard aerobic biodegradability test. Both aniline and phenol attained a biodegradation level higher than 60% in a short time period (<28 days) and thus can be considered as readily biodegradable in denitrifying environments. Nevertheless, the kinetics obtained in the anoxic test were slower than in aerobic conditions, and even suggested the accumulation of intermediate metabolites in the case of phenol. The results of this study indicate that the fate of xenobiotic compounds under anoxic conditions differs from that observed in an oxic environment, and therefore it should be considered by standard biodegradability testing procedures.     

Bioremediation potential of a perchlorate-enriched sewage sludge consortium

The purpose of this work was to explore the reductive bioremediation potential of a perchlorate-enriched facultative anaerobic consortium. Rapid perchlorate reduction and bacterial growth were observed up to 1.84 g l(-1) of perchlorate, but not at 3.82 g l(-1) due to the toxicity. The specific growth rate of the mixed consortium was 0.1 h(-1). The consortium co-reduced perchlorate and nitrate with acetate as e- donor and carbon source. The presence of nitrate slowed down the perchlorate reduction rate. The other e- acceptors utilized include oxygen, chlorate, Cr(VI), and selenate. Over 95% of the 16 mg l(-1) of added Cr(VI) was reduced within 24 h of incubation with a high-density perchlorate-grown consortium. However, the consortium failed to couple growth with reduction of nitrite, sulfate, thiosulfate, and sulfite. During the search for autotrophic perchlorate reduction, many consortia from very diverse natural sources could not use sulfur compounds such as thiosulfate as e- donor.     

Anaerobic degradation of microcrystalline cellulose: kinetics and micro-scale structure evolution

The degradation kinetics and micro-scale structure change of microcrystalline cellulose during anaerobic biodegradation were investigated. A modified Logistic model was established to properly describe the kinetics, which showed good fitness and wide applicability for cellulose degradation. A maximum degradation rate of 0.14 g L⁻¹ h⁻¹ was achieved after cultivating for 51.5 h. This result was in good agreement with the scanning electron microscope and X-ray diffraction analysis. Channels of 400-500 nm size started to occur on the crystalline surface of cellulose at around the inflexion time. Accordingly, the crystallinity significantly decreased at this point, indicating a degradation of the crystalline structure zones by anaerobic bacteria. This study offers direct morphological evidence and quantitative analysis of the biodegradation process of cellulose, and is beneficial to a better understanding of the cellulose degradation mechanism.     

Anaerobic biodegradation of 4-alkylphenols in a paddy soil microcosm supplemented with nitrate

Anaerobic degradation of phenol, p-cresol, 4-n-propylphenol (n-PP), 4-i-propylphenol (i-PP), 4-n-butylphenol (n-BP) and 4-sec-butylphenol (sec-BP) was observed in a paddy soil supplemented with nitrate. We detected the metabolites 4′-hydroxypropiophenone (HPP) from n-PP, 4-i-propenylphenol from i-PP, and 4-(1-butenyl)phenol and 4′-hydroxybutyrophenone (HBP) from n-BP. Compared with the original soils, Betaproteobacteria became predominant in the microcosm during the degradation of phenol and p-cresol whereas no remarkable change was observed in the community degrading propylphenols and butylphenols. The microcosm, however, did not degrade 4-t-butylphenol (t-BP), 4-t-octylphenol (t-OP) and 4-n-octylphenol (n-OP). Paddy soil supplemented with sulfate or iron (III) as electron acceptors did not degrade phenol and 4-alkylphenols with the exception of the degradation of p-cresol in sulfate-reducing conditions. It was demonstrated for the first time that anaerobic microbial degradation of alkylphenols, in a paddy soil supplemented with nitrate as an electron acceptor, occurred via oxidation of the alpha carbon in the alkyl chain.     

Comparative effectiveness of mixed organic substrates to mushroom compost for treatment of mine drainage in passive bioreactors

Bioreactors are one possible best sustainable technology to address the mine-impacted water problems. Several prospective substrates (mushroom compost, cow manure, sawdust, wood chips, and cut rice straw) were characterized for their ability to serve as a source of food and energy for sulfate-reducing bacteria. Twenty bench-scale batch bioreactors were then designed and set up to investigate relative effectiveness of various mixtures of substrates to that of mushroom compost, the most commonly used substrate in field bioreactors, for treating mine drainage with acidic (pH 3) and moderate pH (pH 6). Overall, reactive mixtures showed satisfactory performances in generating alkalinity, reducing sulfate and removing metals (Al>Fe>Mn) (up to 100%) at both pH conditions, for all substrates. The mixture of sawdust and cow manure was found as the most effective whereas the mixture containing 40% cut rice straw gave limited efficiency, suggesting organic carbon released from this substrate is not readily available for biodegradation under anaerobic conditions. The mushroom compost-based bioreactors released significant amount of sulfate, which may raise a more concern upon the start-up of field-scale bioreactors. The correlation between the extent of sulfate reduction and dissolved organic carbon/SO(4)(2-) ratio was weak and this indicates that the type of dissolved organic carbon plays a more important role in sulfate reduction than the absolute concentration and that the ratio is not sensitive enough to properly describe the relative effectiveness of substrate mixtures.     

纳米铁系双金属-微生物体系去除地下水NO-3-N研究

采用不同液相还原法制备纳米Fe0、Fe/Ni和Fe/Cu粒子,将其与反硝化细菌混合应用于地下水NO3--N去除研究。考察3种体系对NO3--N去除速率的影响,并对其脱氮产物及RNA水平上纳米铁系双金属对反硝化细菌的毒性效应进行了分析和讨论。结果表明,9 d内纳米Fe0体系可完全将NO3--N去除,过程中伴随NO2--N先升高后降低的生成趋势,NH 4+-N生成52%;纳米Fe/Ni体系脱氮速率最快,6 d内可将NO 3--N完全去除,几乎未检测到NO 2--N的生成,而NH 4+-N的转化率高达69%;纳米Fe/Cu体系7 d内可将NO3--N去除完全,NH4+-N的生成率降低,仅39%,但是出现33%NO2--N积累。从反应前后反硝化细菌总RNA浓度变化看,3种纳米粒子对反硝化细菌的毒性大小为纳米Fe/Ni﹥纳米Fe/Cu﹥纳米Fe0。     

水质净化高效复合微生态制剂的研制

在室内模拟条件下,采用正交实验的方法对光合细菌、枯草芽孢杆菌和反硝化细菌的复配比例进行了研究,筛选一种用于水产养殖水质净化的高效复合微生态制剂。结果表明,当光合细菌(菌细胞浓度约为2×109CFU/mL)、枯草芽孢杆菌(菌细胞浓度约为8×108CFU/mL)和反硝化细菌(菌细胞浓度约为8×108CFU/mL)按菌液体积比为1∶2∶1进行复配利于水中溶解氧的提高和COD、氨氮、亚硝态氮、硝态氮的降解。验证实验表明,筛选组合各指标均优于商品微生态制剂和空白对照,其中溶解氧含量显著高于商品微生态制剂EM和复合芽孢菌处理,在实验第5天对COD的降解率为95%,显著优于EM和复合芽孢菌处理的66.3%和47.9%,实验第7天对氨氮、亚硝态氮和硝态氮的降解率分别达到70%、89%和56%。     

An anaerobic continuous-flow fixed-bed reactor sustaining a 3-chlorobenzoate-degrading denitrifying population utilizing versatile electron donors and acceptors

An anaerobic continuous-flow fixed-bed column reactor capable of degrading 3-chlorobenzoate (3-CBA) under denitrifying conditions was established, and its rate reached 2.26 mM d(-1). The denitrifying population completely degraded 3-CBA when supplied at 0.1-0.54 mM, but its activity was partly suppressed when 3-CBA was supplied at 0.89 mM. Nitrate was concomitantly consumed throughout the operation of the reactor, the amount of which was similar to or up to 35% higher than the theoretical stoichiometric value that was calculated by assuming that 3-CBA degradation is coupled with denitrification. Batch incubation experiments proved that nitrate is strictly required for 3-CBA degradation in the absence of molecular oxygen. The population also degraded 3-CBA aerobically. Benzoate and 4-CBA were degraded under denitrifying conditions as well as 3-CBA, but 2-CBA was not. Considering that the previously reported denitrifying 3-CBA-degrading cultures do not exhibit 4-CBA degradation under denitrifying conditions, nor aerobic 3-CBA degradation [FEMS Microbiol. Lett. 144 (1996) 213, Appl. Environ. Microbiol. 66 (2000) 3446], the microbial population developed in this experiment was physiologically versatile with respect to the utilization of both electron donors and electron acceptors.     

Mechanisms of electron acceptor utilization: implications for simulating anaerobic biodegradation

Simulation of biodegradation reactions within a reactive transport framework requires information on mechanisms of terminal electron acceptor processes (TEAPs). In initial modeling efforts, TEAPs were approximated as occurring sequentially, with the highest energy-yielding electron acceptors (e.g. oxygen) consumed before those that yield less energy (e.g., sulfate). Within this framework in a steady state plume, sequential electron acceptor utilization would theoretically produce methane at an organic-rich source and Fe(II) further downgradient, resulting in a limited zone of Fe(II) and methane overlap. However, contaminant plumes often display much more extensive zones of overlapping Fe(II) and methane. The extensive overlap could be caused by several abiotic and biotic processes including vertical mixing of byproducts in long-screened monitoring wells, adsorption of Fe(II) onto aquifer solids, or microscale heterogeneity in Fe(III) concentrations. Alternatively, the overlap could be due to simultaneous utilization of terminal electron acceptors. Because biodegradation rates are controlled by TEAPs, evaluating the mechanisms of electron acceptor utilization is critical for improving prediction of contaminant mass losses due to biodegradation. Using BioRedox-MT3DMS, a three-dimensional, multi-species reactive transport code, we simulated the current configurations of a BTEX plume and TEAP zones at a petroleum-contaminated field site in Wisconsin. Simulation results suggest that BTEX mass loss due to biodegradation is greatest under oxygen-reducing conditions, with smaller but similar contributions to mass loss from biodegradation under Fe(III)-reducing, sulfate-reducing, and methanogenic conditions. Results of sensitivity calculations document that BTEX losses due to biodegradation are most sensitive to the age of the plume, while the shape of the BTEX plume is most sensitive to effective porosity and rate constants for biodegradation under Fe(III)-reducing and methanogenic conditions. Using this transport model, we had limited success in simulating overlap of redox products using reasonable ranges of parameters within a strictly sequential electron acceptor utilization framework. Simulation results indicate that overlap of redox products cannot be accurately simulated using the constructed model, suggesting either that Fe(III) reduction and methanogenesis are occurring simultaneously in the source area, or that heterogeneities in Fe(III) concentration and/or mineral type cause the observed overlap. Additional field, experimental, and modeling studies will be needed to address these questions.     

Biodegradation of benzene, toluene, ethylbenzene and xylenes in gas-condensate-contaminated ground-water

The rate and extent of biodegradation of benzene, toluene, ethylbenzene and xylenes (BTEX) in ground-water was studied in samples from a contaminated site which contained total BTEX concentrations of up to 20 000 microg litre(-1). All compounds were rapidly degraded under natural aerobic conditions. Elevation of incubation temperature, supply of organic nutrients or addition of inorganic fertiliser did not increase the rate or extent of biodegradation and it appeared that oxygen supply was the factor limiting BTEX degradation at this site. Attempts to increase the dissolved oxygen concentration in the ground-water by the addition of hydrogen peroxide to give a final concentration of 200 mg litre(-1) resulted in the complete inhibition of biodegradation. No biodegradation occurred under anaerobic conditions except when nitrate was provided as a terminal electron acceptor for microbial respiration. Under denitrifying conditions there was apparent biodegradation of benzene, toluene, ethyl-benzene, m-xylene and p-xylene but o-xylene was not degraded. Degradation under denitrifying conditions occurred at a much slower rate than under oxygenated conditions.