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.

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An experiment was performed in a carbon tetrachloride (CT)- and nitrate-contaminated aquifer at Schoolcraft, MI, to evaluate bioaugmentation with Pseudomonas stutzeri KC, a denitrifying bacterium that degrades CT without producing chloroform (CF). A test section of the aquifer was treated to create pH conditions favorable for KC and then inoculated with culture grown aerobically on site. Activity was sustained with pulses of acetate-amended groundwater, followed by “chase” pulses of acetate-free water. In regions with effective substrate delivery, KC was detected, nitrate levels fell by 85%, pH levels increased, and CT levels decreased by ∼65%, with no significant increase in CF. After 3 weeks, denitrification and CT transformation activity decreased, and KC was no longer detected in groundwater from four wells. Loss of denitrification was attributed to the acetate-free chase. Upon eliminating the chase, CT transformation resumed, and KC was detected, but CF production was also observed, implicating indigenous organisms as agents of transformation. Final sediment analyses indicated 60−88% CT removal, little CF, and persistent KC. This work demonstrates subsurface pH adjustment, subsurface transport of KC, assimilation of KC into the aquifer community, CT removal without CF production after inoculation, and CF formation when KC activity declined.

[1] This paper evaluates the microbial transport and degradation processes associated with carbon tetrachloride (CT) biodegradation in laboratory aquifer columns operated with a pulsed microbial feeding strategy. A seven component reactive transport model based on modified saturation kinetics and on a two-site sorption model was developed to describe the linked physical, chemical, and biological processes involved in CT degradation by Pseudomonas stutzeri KC, a denitrifying bacterium that cometabolically converts CT to harmless end products. After evaluating several expressions for attachment and detachment, we selected a dynamic partitioning model in which strain KC detachment decreases at low substrate concentrations. The resulting model enabled improved understanding of the complex coupled processes operative within our system and enabled us to test a model for field-scale design and transport studies. Batch studies were used to identify initial degradation and microbial transport processes, and constrained optimization methods were used to estimate a set of reaction rates that best describe the column experiment data. The optimal set of parameters for one column provided a reasonable prediction of solute and microbial concentrations in a second column operated under different conditions, providing an initial test of the model. This modeling strategy improved our understanding of biodegradation processes and rates. The CT degradation rate in the columns was lower than values obtained from batch studies, and processes in addition to the growth and decay of strain KC cells (due to native flora) are necessary to describe the observed nitrate consumption.

Pseudomonas stutzeri KC is an aquifer isolate capable of denitrification and cometabolic transformation of carbon tetrachloride (CT) under anoxic conditions. Capillary experiments established that strain KC is chemotactic toward nitrate. A model aquifer column was used to evaluate the effects of motility in the presence of groundwater flow, and a second column was used to isolate motility in the absence of flow. Both columns contained CT-saturated sediments and groundwater containing CT and nitrate. The flow column was inoculated near its upstream end with strain KC, base, acetate, and phosphate and flushed continuously thereafter with contaminated groundwater. Cells migrated through the column at a velocity exceeding that of the groundwater, removing both aqueous and sorbed CT. The no-flow column was inoculated near its midpoint and maintained as a static incubation. Motile KC cells migrated over a 0.3 m distance within 5 days, giving a cell velocity of >5 cm/day. Over 94% of the CT in the column was removed in 26 days. The results support the hypothesis that localized depletion of nitrate during denitrification creates nitrate gradients that trigger a chemotactic response. The results also indicate that motile KC cells can degrade CT.