Field evidence for a protistan role in an organically-contaminated aquifer

The association between protists, bacteria, and dissolved organic carbon (DOC) in an oxygen-depleted, 6 km-long wastewater contaminant plume within a sandy aquifer (Cape Cod, MA) was investigated by comparing abundance patterns along longitudinal and vertical transects and at a control site. Strong linear correlations were observed between unattached bacterial abundance and DOC for much of the upgradient-half of the plume (0.1-2.5 km downgradient from the source) that is characterized by quasi-steady state chemistry. However, a logarithmic decrease was observed between the number of protists supported per mg of DOC and the estimated age of the DOC within the plume. The relatively labile dissolved organic contaminants that characterize the groundwater sampled from the plume < or = 0.1 km downgradient from the contaminant source appeared to indirectly support 3-4 times as many protists (per mg of DOC) as the older, more recalcitrant DOC in the alkylbenzene sulfonate (ABS)-contaminated zone at 3 km downgradient (approximately 30 years travel time). Substantive numbers of protists (>10(4)/cm3) were recovered from suboxic zones of the plume. The higher than expected ratios of protists to unattached bacteria (10 to 100:1) observed in much of the plume suggest that protists may be grazing upon both surface-associated and unattached bacterial communities to meet their nutritional requirements. In closed bottle incubation experiments, the presence of protists caused an increase in bacterial growth rate, which became more apparent at higher amendments of labile DOC (3-20 mgC/L). The presence of protists resulted in an increase in the apparent substrate saturation level for the unattached bacterial community, suggesting an important role for protists in the fate of more-labile aquifer organic contaminants.     

Stable carbon isotope evidence for intrinsic bioremediation of tetrachloroethene and trichloroethene at area 6, Dover Air Force Base

Area 6 at Dover Air Force Base (Dover, DE) has been the location of an in-depth study by the RTDF (Remediation Technologies Development Forum Bioremediation of Chlorinated Solvents Action Team) to evaluate the effectiveness of natural attenuation of chlorinated ethene contamination in groundwater. Compound-specific stable carbon isotope measurements for dissolved PCE and TCE in wells distributed throughout the anaerobic portion of the plume confirm that stable carbon isotope values are isotopically enriched in 13C consistent with the effects of intrinsic biodegradation. During anaerobic microbial reductive dechlorination of chlorinated hydrocarbons, the light (12C) versus heavy isotope (13C) bonds are preferentially degraded, resulting in isotopic enrichment of the residual contaminant in 13C. To our knowledge, this study is the first to provide definitive evidence for reductive dechlorination of chlorinated hydrocarbons at a field site based on the delta13C values of the primary contaminants spilled at the site, PCE and TCE. For TCE, downgradient wells show delta13C values as enriched as -18.0/1000 as compared to delta13C values for TCE in the source zone of -25.0 to -26.0/1000. The most enriched delta13C value on the site was observed at well 236, which also contains the highest concentrations of cis-DCE, VC, and ethene, the daughter products of reductive dechlorination. Stable carbon isotope signatures are used to quantify the relative extent of biodegradation between zones of the contaminant plume. On the basis of this approach, it is estimated that TCE in downgradient well 236 is more than 40% biodegraded relative to TCE in the proposed source area.     

Steroid estrogens, nonylphenol ethoxylate metabolites, and other wastewater contaminants in groundwater affected by a residential septic system on Cape Cod, MA

Septic systems serve approximately 25% of U.S. households and may be an important source of estrogenic and other organic wastewater contaminants (OWC) to groundwater. We monitored several estrogenic OWC, including nonylphenol (NP), nonylphenol mono- and diethoxycarboxylates (NP1EC and NP2EC), the steroid hormones 17beta-estradiol (E2), estrone (E1) and their glucuronide and sulfate conjugates, and other OWC such as methylene blue active substances (MBAS), caffeine and its degradation product paraxanthine, and two fluorescent whitening agents in a residential septic system and in downgradient groundwater. E1 and E2 were present predominantly as free estrogens in groundwater, and near-source groundwater concentrations of all OWC were highest in the suboxic to anoxic portion of the wastewater plume, where concentrations of most OWC were similar to those observed in the septic tank on the same day. NP and NP2EC were up to 6- to 30-fold higher, and caffeine and paraxanthine were each 60-fold lower than septic tank concentrations, suggesting net production and removal, respectively, of these constituents. At the most shallow, oxic depth, concentrations of all OWC except for NP2EC were substantially lower than in the tank and in deeper wells. Yet boron, specific conductance, and the sum of nitrate-and ammonia-nitrogen were highest at this shallow depth, suggesting preferential losses of OWC along the more oxic flow lines. As far as 6.0 m downgradient, concentrations of many OWC were within a factor of 2 of near-source concentrations. The results suggest that there is the potential for migration of these OWC, which are unregulated and not routinely monitored, in groundwater.     

GeoChip-based analysis of microbial functional gene diversity in a landfill leachate-contaminated aquifer

The functional gene diversity and structure of microbial communities in a shallow landfill leachate-contaminated aquifer were assessed using a comprehensive functional gene array (GeoChip 3.0). Water samples were obtained from eight wells at the same aquifer depth immediately below a municipal landfill or along the predominant downgradient groundwater flowpath. Functional gene richness and diversity immediately below the landfill and the closest well were considerably lower than those in downgradient wells. Mantel tests and canonical correspondence analysis (CCA) suggested that various geochemical parameters had a significant impact on the subsurface microbial community structure. That is, leachate from the unlined landfill impacted the diversity, composition, structure, and functional potential of groundwater microbial communities as a function of groundwater pH, and concentrations of sulfate, ammonia, and dissolved organic carbon (DOC). Historical geochemical records indicate that all sampled wells chronically received leachate, and the increase in microbial diversity as a function of distance from the landfill is consistent with mitigation of the impact of leachate on the groundwater system by natural attenuation mechanisms.     

New evaluation scheme for two-dimensional isotope analysis to decipher biodegradation processes: application to groundwater contamination by MTBE

Compound-specific analysis of stable carbon and hydrogen isotopes was used to assess the fate of the gasoline additive methyl tert-butyl ether (MTBE) and its major degradation product tert-butyl alcohol (TBA) in a groundwater plume at an industrial disposal site. We present a novel approach to evaluate two-dimensional compound-specific isotope data with the potential to identify reaction mechanisms and to quantify the extent of biodegradation at complex field sites. Due to the widespread contaminant plume, multiple MTBE sources, the presence of numerous other organic pollutants, and the complex biogeochemical and hydrological regime atthe site, a traditional mass balance approach was not applicable. The isotopic composition of MTBE steadily changed from the source regions along the major contaminant plume (-26.4% to +40.0% (carbon); -73.1% to +60.3% (hydrogen)) indicating substantial biodegradation. Constant carbon isotopic signatures of TBA suggest the absence of TBA degradation at the site. Published carbon and hydrogen isotope fractionation data for biodegradation of MTBE under oxic and anoxic conditions, respectively, were examined and used to determine both the nature and the extent of in-situ biodegradation along the plume(s). The coupled evaluation of two-dimensional compound-specific isotope data explained both carbon and hydrogen fractionation data in a consistent way and indicate anaerobic biodegradation of MTBE along the entire plume. A novel scheme to reevaluate empiric isotopic enrichment factors (epsilon) in terms of theoretically based intrinsic carbon (12k/13k) and hydrogen (1k/2k) kinetic isotope effects (KIE) is presented. Carbon and hydrogen KIE values, calculated for different potential reaction mechanisms, imply that anaerobic biodegradation of MTBE follows a SN2-type reaction mechanism. Furthermore, our data suggest that additional removal process(es) such as evaporation contributed to the overall MTBE removal along the plume, a phenomenon that might be significant also for other field sites at tropic or subtropic climates with elevated groundwater temperatures (25 degrees C).     

Origin of a mixed brominated ethene groundwater plume: contaminant degradation pathways and reactions

On the basis of a combination of laboratory microcosm experiments, column sorption experiments, and the current spatial distribution of groundwater concentrations, the origin of a mixed brominated ethene groundwater plume and its degradation pathway were hypothesized. The contaminant groundwater plume was detected downgradient of a former mineral processing facility, and consisted of tribromoethene (TriBE), cis-1,2-dibromoethene (c-DBE), trans-1,2-dibromoethene (t-DBE), and vinyl bromide (VB). The combined laboratory and field data provided strong evidence that the origin of the mixed brominated ethene plume was a result of dissolution of the dense non-aqueous-phase liquid 1,1,2,2-tetrabromoethane (TBA) atthe presumed source zone, which degraded rapidly (half-life of 0.2 days) to form TriBE in near stoichiometric amounts. TriBE then degraded (half-life of 96 days) to form c-DBE, t-DBE, and VB via a reductive debromination degradation pathway. Slow degradation of c-DBE (half-life >220 days), t-DBE (half-life 220 days), and VB (half-life >220 days) coupled with their low retardation coefficients (1.2, 1.2, and 1.0 respectively) resulted in the formation of an extensive mixed brominated ethene contaminant plume. Without this clearer understanding of the mechanism for TBA degradation, the origin of the mixed brominated ethene groundwater contamination could have been misinterpreted, and inappropriate and ineffective source zone and groundwater remediation techniques could be applied.     

Isotopic tracing of landfill leachates and pollutant lead mobility in soil and groundwater

Here we provide evidence of the capability of stable lead isotopes to trace landfill leachate in a shallow groundwater. The municipal landfill we have investigated is located in southeastern France. It has no bottom liner, and wastes are placed directly on the ground. Stable lead isotopes allow the characterization of this landfill leachate signature (206Pb/207Pb = 1.189 +/- 0.004) that is clearly different from that of the local atmosphere (206Pb/207Pb = 1.150 +/- 0.006) and crustal lead (206Pb/207Pb = 1.200 +/- 0.005). Piezometers located in the direct vicinity of the landfill generally display this contaminant imprint. The landfill plume is monitored up to 1000 m downgradient of the landfill, in very good agreement with evaluation from chloride concentration. Meanwhile, 206Pb/207Pb ratios measured at a piezometer located 4600 m downgradient of the landfill suggest a contamination by the landfill plume. This result shows that the complexity of a pollutant plume dispersion in this shallow groundwater system requires several independent tracers to clearly resolve origin and transport pathways for contaminants. Furthermore, seasonal rainfall variation for this Mediterranean mixed Quaternary alluvion reservoir and the use of KCl fertilizers might favor an efficient remobilization of atmospheric lead in plowed soils and its transfer into groundwater as shown by lead isotope systematics.     

Pump-and-treat remediation of chlorinated solvent contamination at a controlled field-experiment site

Pump-and-treat (P&T) remediation and associated concentration tailing are investigated at the field scale in a mildly heterogeneous sandy aquifer through the extraction of dissolved chlorinated solvent plumes that had developed over 475 d from a multicomponent dense nonaqueous-phase liquid (DNAPL) source intentionally emplaced in the aquifer at the Borden (ON) research site. Extraction was accomplished via a source-containment well located 25 m from the source and two further downgradient plume-centerline wells to remove the advancing high-concentration dissolved plumes. The 550 days of detailed P&T field data demonstrated the following: remediation, albeit slowly, of the leading 25-60 m plume section to around typical drinking water standard concentrations; concentration tailing (reduction) over 4 orders of magnitude in the plume; a steady-state concentration "plateau" in the source-containment well capturing the steadily dissolving DNAPL source; influences of extraction rate changes (concentration rebounds); and, lengthy tailing from inter-well stagnation-zone areas. Much of the contaminant behavior during the P&T appeared to be "ideal" in the sense that with appropriate specification of the source term and pumping regime, it was reasonably predicted by 3-dimensional numerical model (HydroGeoSphere) simulations that assumed ideal (macrodispersion, linear sorption, etc.) transport. Supporting lab studies confirmed nonideal sorption was, however, important at the point sample scale with enhanced PCE (tetrachloroethene) sorption to low- and high-permeability strata and moderate nonlinear and competitive sorption influences. Although there was limited evidence of nonideal tailing contributions to the field data (underprediction of some tailing curve gradients), such contributions to P&T tailing were not easily discerned and appeared to play a relatively minor role within the mildly heterogeneous aquifer studied.     

Anaerobic methane oxidation in a landfill-leachate plume

The alluvial aquifer adjacent to Norman Landfill, OK, provides an excellent natural laboratory for the study of anaerobic processes impacting landfill-leachate contaminated aquifers. We collected groundwaters from a transect of seven multilevel wells ranging in depth from 1.3 to 11 m that were oriented parallel to the flow path. The center of the leachate plume was characterized by (1) high alkalinity and elevated concentrations of total dissolved organic carbon, reduced iron, and methane, and (2) negligible oxygen, nitrate, and sulfate concentrations. Methane concentrations and stable carbon isotope (delta13C) values suggest anaerobic methane oxidation was occurring within the plume and at its margins. Methane delta13C values increased from about -54 per thousand near the source to > -10 per thousand downgradient and at the plume margins. The isotopic fractionation associated with this methane oxidation was -13.6+/-1.0 per thousand. Methane 13C enrichment indicated that 80-90% of the original landfill methane was oxidized over the 210-m transect. First-order rate constants ranged from 0.06 to 0.23 per year, and oxidation rates ranged from 18 to 230 microM/y. Overall, hydrochemical data suggest that a sulfate reducer-methanogen consortium may mediate this methane oxidation. These results demonstrate that natural attenuation through anaerobic methane oxidation can be an important sink for landfill methane in aquifer systems.     

Organic matter and modeling redox reactions during river bank filtration in an alluvial aquifer of the Lot River, France

A 3 year study of the infiltration of Lot River water into a well field located in an adjacent gravel and clay alluvial aquifer was conducted to assess the importance of organic matter regarding the redox processes which influence groundwater quality in a positive (denitrification) or negative (Mn dissolution) manner. Chloride was used to quantify the mixing of river water with groundwater. According to modeling with PHREEQC, the biodegradation of the infiltrated dissolved organic carbon (DOCi) is not sufficient to explain the observed consequences of the redox reactions (dissolved O2 depletion, denitrification, Mn dissolution). Another electron donor source must therefore be involved: we propose solid organic carbon (SOC) as a likely candidate, if made available for degradation by prior hydrolysis. Its contribution to redox reactions could be significant (30-80% of the total organic carbon consumed by redox reactions during river bank filtration). We show here also that even though the first few meters of infiltration are highly reactive, significant redox reactions can take place further in the aquifer, possibly affecting groundwater quality away from the river bank.     

Evidence of enantioselective degradation of alpha-hexachlorocyclohexane in groundwater

In the fall of 2000, 34 groundwater samples were collected from beneath an active pesticide reformulating and packaging facility in coastal northeastern Florida to measure the enantiomer fractions (EFs) of alpha-hexachlorocyclohexane (alpha-HCH) as an indicator of biodegradation of this chlorinated pesticide in groundwater. Concentrations of alpha-HCH as high as 500 microg/L were observed beneath the historical source area and decreased with distance downgradient. Seventy-eight percent of the EF values were greater than 0.504 and ranged up to 0.890, indicating that the (-)-alpha-HCH enantiomer is preferentially degraded relative to the (+)-alpha-HCH enantiomer at this site. Samples taken from the groundwater that flows north from the historical disposal facility to a local discharge point at a creek did not indicate enantioselective degradation (EF values ranged from 0.495 to 0.512). The acidity (pH 3.7-4.6) and short flow path to the creek for this lobe of the groundwater plume likely preclude biodegradation of alpha-HCH. In contrast, the neutral lobe of the groundwater plume, which flows eastward from the historical source area, demonstrated enantioselective degradation (EF values ranged from 0.500 to 0.890 and increased with distance from the source area). Groundwater conditions beneath this portion of the site are conducive to biodegradation of HCH owing to anaerobic reducing conditions and lengthy travel times, and the chiral signatures for alpha-HCH provide evidence that biological degradation is occurring beneath this portion of the site.     

Hydrogen isotopic enrichment: an indicator of biodegradation at a petroleum hydrocarbon contaminated field site

Compound-specific carbon and hydrogen isotope analysis was used to investigate biodegradation of benzene and ethylbenzene in contaminated groundwater at Dow Benelux BV industrial site. delta13C values for dissolved benzene and ethylbenzene in downgradient samples were enriched by up to 2+/-0.5 per thousand, in 13C, compared to the delta13C value of the source area samples. delta2H values for dissolved benzene and ethylbenzene in downgradient samples exhibited larger isotopic enrichments of up to 27+/-5 per thousand for benzene and up to 50+/-5 per thousand for ethylbenzene relative to the source area. The observed carbon and hydrogen isotopic fractionation in downgradient samples provides evidence of biodegradation of both benzene and ethylbenzene within the study area at Dow Benelux BV. The estimated extents of biodegradation of benzene derived from carbon and hydrogen isotopic compositions for each sample are in agreement, supporting the conclusion that biodegradation is the primary control on the observed differences in carbon and hydrogen isotope values. Combined carbon and hydrogen isotope analyses provides the ability to compare biodegradation in the field based on two different parameters, and hence provides a stronger basis for assessment of biodegradation of petroleum hydrocarbon contaminants.     

Applicability of stable isotope fractionation analysis for the characterization of benzene biodegradation in a BTEX-contaminated aquifer

In recent years the analysis of stable isotope fractionation has increasingly been used for characterizing and quantifying biodegradation of contaminants in aquifers. The correlation of carbon and hydrogen isotope signatures of benzene in a BTEX-contaminated aquifer located in the area of a former hydrogenation plant gave indications that biodegradation mainly occurred under anoxic conditions. This finding was consistent with the investigation of hydrogeochemical conditions within the aquifer. Furthermore, the biodegradation of benzene was calculated by changes in carbon isotope signatures using the Rayleigh-equation-streamline approach. Since contaminant concentrations can be also affected by nonisotope-fractionating abiotic processes such as dilution, volatilization, or irreversible sorption to the aquifer matrix, the Rayleigh-equation-streamline approach was adjusted for scenarios assuming that biodegradation and abiotic processes occur either consecutively or simultaneously along a groundwater flow path between contaminant source and sampling well. The results of the scenarios differed significantly, indicating that an abiotic process (typically dilution) causes a decrease in benzene concentration within the investigated aquifer transect. This comparison of results derived from the different scenarios can help to identify whether biodegradation is the predominant process for decrease in contaminant concentration. However, for a proper quantification of biodegradation, the temporal sequence between biodegradation and dilution needs to be known. The uncertainty associated with the quantification of pollutant biodegradation by the Rayleigh-equation-streamline approach increases when nonisotope-fractionating abiotic processes cause a significant decrease in contaminant concentrations.     

Modeling the dynamics of fermentation and respiratory processes in a groundwater plume of phenolic contaminants interpreted from laboratory- to field-scale

A biodegradation model with consecutive fermentation and respiration processes, developed from microcosm experiments and simulated mathematically with microbial growth kinetics, has been implemented into a field-scale reactive transport model of a groundwater plume of phenolic contaminants. Simulation of the anaerobic plume core with H2 and acetate as intermediate products of biodegradation allows the rates and parameter values forfermentation processes and individual respiratory terminal electron accepting processes (TEAPS) to be estimated using detailed, spatially discrete, hydrochemical field data. The modeling of field-scale plume development includes consideration of microbial acclimatization, substrate toxicity toward degradation, bioavailability of mineral oxides, and adsorption of biogenic Fe(ll) species in the aquifer, identified from complementary laboratory process studies. The results suggest that plume core processes, particularly fermentation and Fe(lll)-reduction, are more important for degradation than previously thought, possibly with a greater impact than plume fringe processes (aerobic respiration, denitrification, and SO4-reduction). The accumulation of acetate as a fermentation product within the plume contributes significantly to the mass balance for carbon. These results demonstrate the value of quantifying fermentation products within organic contaminant plumes and strongly suggest that the conceptual model selected for reactive processes plays a dominant role in the quantitative assessment of risk reduction by naturally occurring biodegradation processes.     

Reductions in contaminant mass discharge following partial mass removal from DNAPL source zones

Although in situ remediation technologies have been used to aggressively treat dense nonaqueous phase liquid (DNAPL) source zones, complete contaminant removal or destruction is rarely achieved. To evaluate the effects of partial source zone mass removal on dissolved-phase contaminant flux, four experiments were conducted in a two-dimensional aquifer cell that contained a tetrachloroethene (PCE) source zone and down-gradient plume region. Initial source zone PCE saturation distributions, quantified using a light transmission system, were expressed in terms of a ganglia-to-pool ratio (GTP), which ranged from 0.16 (13.8% ganglia) to 1.6 (61.5% ganglia). The cells were flushed sequentially with a 4% (wt.) Tween 80 surfactant solution to achieve incremental PCE mass removal, followed by water flooding until steady-state mass discharge and plume concentrations were established. In all cases, the GTP ratio decreased with increasing mass removal, consistent with the observed preferential dissolution of PCE ganglia and persistence of high-saturation pools. In the ganglia-dominated system (GTP = 1.6), greater than 70% mass removal was required before measurable reductions in plume concentrations and mass discharge were observed. For pool-dominated source zones (GTP < 0.3), substantial reductions (>50%) in mass discharge were realized after only 50% mass removal.     

Anaerobic biodegradation of ethylene dibromide and 1,2-dichloroethane in the presence of fuel hydrocarbons

Field evidence from underground storage tank sites where leaded gasoline leaked indicates the lead scavengers 1,2-dibromoethane (ethylene dibromide, or EDB) and 1,2-dichloroethane (1,2-DCA) may be present in groundwater at levels that pose unacceptable risk. These compounds are seldom tested for at UST sites. Although dehalogenation of EDB and 1,2-DCA is well established, the effect of fuel hydrocarbons on their biodegradability under anaerobic conditions is poorly understood. Microcosms (2 L glass bottles) were prepared with soil and groundwater from a UST site in Clemson, South Carolina, using samples collected from the source (containing residual fuel) and less contaminated downgradient areas. Anaerobic biodegradation of EDB occurred in microcosms simulating natural attenuation, but was more extensive and predictable in treatments biostimulated with lactate. In the downgradient biostimulated microcosms, EDB decreased below its maximum contaminant level (MCL) (0.05 microg/L) at a first order rate of 9.4 +/- 0.2 yr(-1). The pathway for EDB dehalogenation proceeded mainly by dihaloelimination to ethene in the source microcosms, while sequential hydrogenolysis to bromoethane and ethane was predominant in the downgradient treatments. Biodegradation of EDB in the source microcosms was confirmed by carbon specific isotope analysis, with a delta13C enrichment factor of -5.6 per thousand. The highest levels of EDB removal occurred in microcosms that produced the highest amounts of methane. Extensive biodegradation of benzene, ethylbenzene, toluene and ortho-xylene was also observed in the source and downgradient area microcosms. In contrast, biodegradation of 1,2-DCA proceeded at a considerably slower rate than EDB, with no response to lactate additions. The slower biodegradation rates for 1,2-DCA agree with field observations and indicate that even if EDB is removed to below its MCL, 1,2-DCA may persist.     

Destruction efficiencies and dynamics of reaction fronts associated with the permanganate oxidation of trichloroethylene

Although potassium permanganate (KMnO4) flushing is commonly used to destroy chlorinated solvents in groundwater, many of the problems associated with this treatment scheme have not been examined in detail. We conducted a KMnO4 flushing experiment in a large sand-filled flow tank (L x W x D = 180 cm x 60 cm x 90 cm) to remove TCE emplaced as a DNAPL in a source zone. The study was specifically designed to investigate cleanup progress and problems of pore plugging associated with the dynamics of the solid-phase reaction front (i.e., MnO2) using chemical and optical monitoring techniques. Ambient flow through the source zone formed a plume of dissolved TCE across the flow tank. The volume and concentration of TCE plume diminished with time because of the in situ oxidation of the DNAPL source. The migration velocity of the MnO2 reaction front decreased with time, suggesting that the kinetics of the DNAPL oxidation process became diffusion-controlled because of the pore plugging. A mass balance calculation indicated that only approximately 18% of the total applied KMnO4 (MnO4- = 1250 mg/ L) participated in the oxidation reaction to destroy approximately 41% of emplaced TCE. Evidently, the efficiency of KMnO4 flushing scheme diminished with time due to pore plugging by MnO2 and likely CO2, particularly in the TCE source zone. In addition, the excess KMnO4 used for flushing may cause secondary aquifer contamination. One needs to be concerned about the efficacy of KMnO4 flushing in the field applications. Development of a new approach that can provide both contaminant destruction and plugging/ MnO4- control is required.     

Trace metal levels in uncontaminated groundwater of a coastal watershed: importance of colloidal forms

Groundwater and surface water were collected using trace metal clean techniques from the upper glacial aquifer of West Neck Bay (Shelter Island) in eastern Long Island, NY, during the late spring and summer of 1999. The collection sites on Shelter Island are located in an area that is primarily residential and believed to have uncontaminated groundwater. Ultrafiltration was used to size-fractionate the dissolved (<0.45 microm) fraction into colloidal (1 kDa - 0.45 microm) and low molecular weight (<1 kDa) size pools. These fractions were analyzed for trace metals (Al, Ag, Cd, Cu, Mn, Pb, and Zn), organic carbon, and inorganic nutrients (NH4, NO3, PO4). The levels of metals and organic carbon in the groundwater were as low as those found in the open ocean, far removed from anthropogenic inputs. These findings corroborate the need to apply trace metal clean techniques in the determination of metal levels in uncontaminated groundwater. A significant fraction of dissolved metals (22-96%) and organic carbon (approximately 40%) in the groundwater and in surface waters of the Bay was found to be associated with colloids. The significance of the metal association with the colloidal fraction decreased in the order of Al > Cu > Ag > Zn = Cd = Mn and appeared to be dependent on the affinities of these metals for humic substances. In contrast, NO3 and NH4 were found to be almost entirely (approximately 98-99%) in the low molecular weight size fraction. Metal/aluminum and metal/carbon ratios measured in the colloids were similar to those reported for humic substances and significantly different from those of soils. This suggests that colloidal particles might originate from humic materials as opposed to purely inorganic minerals. These results indicate the need to consider the colloidal fraction in the fate and mobility of metals in groundwater and that, despite the low levels of organic matter (<50 microM of DOC) measured in groundwater, some groundwater colloids appear to be organic in nature.     

Microbial incorporation of 13C-labeled acetate at the field scale: detection of microbes responsible for reduction of U(VI)

A field-scale acetate amendment experiment was performed in a contaminated aquifer at Old Rifle, CO to stimulate in situ microbial reduction of U(VI) in groundwater. To evaluate the microorganisms responsible for microbial uranium reduction during the experiment, 13C-labeled acetate was introduced into well bores via bio-traps containing porous activated carbon beads (Bio-Sep). Incorporation of the 13C from labeled acetate into cellular DNA and phospholipid fatty acid (PLFA) biomarkers was analyzed in parallel with geochemical parameters. An enrichment of active sigma-proteobacteria was demonstrated in downgradient monitoring wells: Geobacter dominated in wells closer to the acetate injection gallery, while various sulfate reducers were prominent in different downgradient wells. These results were consistent with the geochemical evidence of Fe(III), U(VI), and SO(4)2- reduction. PLFA profiling of bio-traps suspended in the monitoring wells also showed the incorporation of 13C into bacterial cellular lipids. Community composition of downgradient monitoring wells based on quinone and PLFA profiling was in general agreement with the 13C-DNA result. The direct application of 13C label to biosystems, coupled with DNA and PLFA analysis,     

Methyl tert-butyl ether biodegradation by indigenous aquifer microorganisms under natural and artificial oxic conditions

Microbial communities indigenous to a shallow groundwater system near Beaufort, SC, degraded milligram per liter concentrations of methyl tert-butyl ether (MTBE) under natural and artificial oxic conditions. Significant MTBE biodegradation was observed where anoxic, MTBE-contaminated groundwater discharged to a concrete-lined ditch. In the anoxic groundwater adjacent to the ditch, concentrations of MTBE were > 1 mg/L. Where groundwater discharge occurs, dissolved oxygen (DO) concentrations beneath the ditch exceeded 1.0 mg/Lto a depth of 1.5 m, and MTBE concentrations decreased to <1 microg/L prior to discharge. MTBE mass flux calculations indicate that 96% of MTBE mass loss occurs in the relatively small oxic zone prior to discharge. Samples of a natural microbial biofilm present in the oxic zone beneath the ditch completely degraded [U-14C]MTBE to [14C]CO2 in laboratory liquid culture studies, with no accumulation of intermediate compounds. Upgradient of the ditch in the anoxic, MTBE- and BTEX-contaminated aquifer, addition of a soluble oxygen release compound resulted in oxic conditions and rapid MTBE biodegradation by indigenous microorganisms. In an observation well located closest to the oxygen addition area, DO concentrations increased from 0.4 to 12 mg/L in <60 days and MTBE concentrations decreased from 20 to 3 mg/L. In the same time period at a downgradient observation well, DO increased from <0.2 to 2 mg/L and MTBE concentrations decreased from 30 to <5 mg/L. These results indicate that microorganisms indigenous to the groundwater system at this site can degrade milligram per liter concentrations of MTBE under natural and artificial oxic conditions.