An integrated evaluation of bioenhanced in situ LNAPL dissolution.

Performance evaluation of in situ bioremediation processes in the field is difficult due to uncertainty created by matrix and contaminant heterogeneity, inaccessibility to direct observation, expense of sampling, and limitations of some measurements. The goal of this research was to develop a strategy for evaluating in situ bioremediation of light nonaqueous-phase liquid (LNAPL) contamination and demonstrating the occurrence of bioenhanced LNAPL dissolution by: (1) integrating a suite of analyses into a rational evaluation strategy; and (2) demonstrating the strategy's application in intermediate-scale flow-cell (ISFC) experiments simulating an aquifer contaminated with a pool of LNAPL (naphthalene dissolved in dodecane). Two ISFCs were operated to evaluate how the monitored parameters changed between a "no bioremediation" scenario and an "intrinsic in situ bioremediation" scenario. Key was incorporating different measures of microbial activity and contaminant degradation relevant to bioremediation: contaminant loss; consumption of electron acceptors; and changes in total alkalinity, pH, dissolved total inorganic carbon, carbon-stable isotopes, microorganisms, and intermediate metabolites. These measurements were integrated via mass-flux modeling and mass-balance analyses to document that in situ biodegradation of naphthalene was strongly accelerated in the "intrinsic in situ bioremediation" scenario versus "no bioremediation." Furthermore, the integrated strategy provided consistent evidence of bioenhancement of LNAPL dissolution through intrinsic bioremediation by a factor of approximately 2 due to the biodegradation of the naphthalene near the pool/water interface.

Predictions of bioenhancement of nonaqueous phase liquid ganglia dissolution using first- and zero-order biokinetic models.

The bioenhanced dissolution of nonaqueous phase liquid (NAPL) contaminants that occurs as a result of an increased concentration gradient is influenced by several factors, including the biokinetics. This is important because available data suggest that at typical NAPL source zone concentrations, descriptions of dissolution bioenhancement may require kinetic expressions ranging from first- to zero-order. In this work, an analytical model for the bioenhancement factor, E, is developed for NAPL ganglia dissolution with zero-order kinetics, and compared to a model for E with first-order kinetics. The models are analyzed and an illustrative example is provided to demonstrate the importance of using the correct biokinetics when estimating the potential magnitude of the bioenhancement of NAPL ganglia dissolution.

Laboratory-scale in situ bioremediation in heterogeneous porous media: biokinetics-limited scenario.

Subsurface heterogeneities influence interfacial mass-transfer processes and affect the application of in situ bioremediation by impacting the availability of substrates to the microorganisms. However, for difficult-to-degrade compounds, and/or cases with inhibitory biodegradation conditions, slow biokinetics may also limit the overall bioremediation rate, or be as limiting as mass-transfer processes. In this work, a quantitative framework based on a set of dimensionless coefficients was used to capture the effects of the competing interfacial and biokinetic processes and define the overall rate-limiting process. An integrated numerical modeling and experimental approach was used to evaluate application of the quantitative framework for a scenario in which slow-biokinetics limited the overall bioremediation rate of a polycyclic aromatic hydrocarbon (naphthalene). Numerical modeling was conducted to simulate the groundwater flow and naphthalene transport and verify the system parameters, which were used in the quantitative framework application. The experiments examined the movement and biodegradation of naphthalene in a saturated, heterogeneous intermediate-scale flow cell with two layers of contrasting hydraulic conductivities. These experiments were conducted in two phases: Phase I, simulating an inhibited slow biodegradation; and Phase II, simulating an engineered bioremediation, with system perturbations selected to enhance the slow biodegradation rate. In Phase II, two engineered perturbations to the system were selected to examine their ability to enhance in situ biodegradation. In the first perturbation, nitrogen and phosphorus in excess of the required stoichiometric amounts were spiked into the influent solution to mimic a common remedial action taken in the field. The results showed that this perturbation had a moderate positive impact, consistent with slow biokinetics being the overall rate-limiting process. However, the second perturbation, which was to alleviate inhibition and increase the biodegradation rate, enhanced the overall biotransformation rate to a greater degree.

A quantitative framework for understanding complex interactions between competing interfacial processes and in situ biodegradation.

In situ bioremediation of contaminated groundwater is made technologically challenging by the physically, chemically, and biologically heterogeneous subsurface environment. Subsurface heterogeneities are important because of influences on interfacial mass transfer processes that impact the availability of substrates to microorganisms. The goal of this study was to perform a "proof-of-concept" evaluation of the utility of a quantitative framework based on a set of dimensionless coefficients for evaluating the effects of competing physicochemical interfacial and biokinetic processes at the field scale. First, three numerical modeling experiments were completed, demonstrating how the framework can be used to identify the rate-limiting process for the overall bioremediation rate, and to predict what engineered enhancements will alleviate the rate-limiting process. Baseline conditions for each scenario were established to examine intrinsic biodegradation with a given rate-limiting process (either dispersion, biokinetics, or sorption). Then different engineering treatments were examined. In each case, the treatment predicted to be appropriate for addressing the overall rate-limiting process based on the quantitative framework alleviated the limitation more successfully, and enhanced the in situ biodegradation rate more than the alternative enhancements. Second, the quantitative framework was applied to a series of large-scale laboratory and field-scale experiments, using reported parameter estimates to calculate the relevant dimensionless coefficients and predict the rate-limiting process(es). Observations from the studies were then used to evaluate those predictions.

Effects of temperature on bacterial transport and destruction in bioretention media: field and laboratory evaluations.

Microbial activities are significantly influenced by temperature. This study investigated the effects of temperature on the capture and destruction of bacteria from urban stormwater runoff in bioretention media using 2-year field evaluations coupled with controlled laboratory column studies. Field data from two bioretention cells show that the concentration of indicator bacteria (fecal coliforms and Escherichia coli) was reduced during most storm events, and that the probability of meeting specific water quality criteria in the discharge was increased. Indicator bacteria concentration in the input flow typically increased with higher daily temperature. Although bacterial removal efficiency was independent of temperature in the field and laboratory, column tests showed that bacterial decay coefficients in conventional bioretention media (CBM) increase exponentially with elevated temperature. Increases in levels of protozoa and heterotrophic bacteria associated with increasing temperature appear to contribute to faster die-off of trapped E. coli in CBM via predation and competition.

The capture and destruction of Escherichia coli from simulated urban runoff using conventional bioretention media and iron oxide-coated sand.

The performance, sustainability, and mechanisms of bacterial removal from stormwater runoff by bioretention systems are poorly understood. The potential for removal of microorganisms in bioretention systems was evaluated using column studies and simulated urban stormwater runoff. Conventional bioretention media (CBM) removed 82% of Escherichia coli O157:H7 strain B6914 cells; iron-oxide coated sand (IOCS) significantly enhanced capture, with 99% efficiency. This improvement possibly was because of the greater positive surface charge and roughness, of the IOCS. Trapped strain B6914 cells decayed more rapidly in CBM, however, with more than 99.98% die-off within one week compared with the IOCS in which approximately 48% of trapped cells survived. Predation and competition from native microorganisms in CBM were verified to play a dominant role in rapid destruction of trapped strain B6914. In particular, protozoan grazing appeared to play an important role, with the die-off of trapped B6914 increasing with increasing concentrations of protozoa.

Modeling the effects of microbial competition and hydrodynamics on the dissolution and detoxification of dense nonaqueous phase liquid contaminants.

The real significance and engineering potential for bioenhanced dissolution of chlorinated ethene dense nonaqueous phase liquid (DNAPL) contaminants are currently not well understood, in part because they can be influenced by a complex set of factors, including microbial competition for growth substrates. Mathematical simulations were performed to evaluate the effects of competition between Dehalococcoides ethenogenes and Desulfuromonas michiganensis for the electron acceptor tetrachloroethene (PCE) on the distribution of dehalorespirers, PCE dissolution, and the extent of PCE detoxification. The modeling results demonstrate that the outcome of competition between populations for growth substrates can have a significant impact on bioenhancement and, thus, on DNAPL source zone longevity and identify the key factors in determining the outcome of competition and its effects on DNAPL dissolution. The potential for bioenhancement is greatest at lower groundwater velocities. At higher velocities, kinetic properties play a key role in determining which population dominates and where, and the amount of bioenhancement that is realized. Engineered bioremediation techniques that maintain multiple dehalorespiring populations may offer the best approach for optimizing the twin cleanup goals of reduced source zone longevity and complete detoxification while maximizing the utilization of added electron donors.

In situ bioremediation in heterogeneous porous media: dispersion-limited scenario.

A quantitative framework based on a set of dimensionless numbers was developed to capture the effects of competing interfacial and biokinetic processes and define limits on the application of in situ bioremediation. An integrated numerical modeling and experimental approach was utilized to evaluate the quantitative framework. Experiments were conducted to examine the transport and biodegradation of naphthalene in a saturated, heterogeneous intermediate-scale flow cell with two layers of contrasting hydraulic conductivities. The experiments were carried out in two phases: Phase I, simulating intrinsic biodegradation; and Phase II, simulating an engineered in situ bioremediation. In Phase I, dispersion was identified as the overall rate-limiting process based on the proposed quantitative framework. Two engineered perturbations to the system were selected in Phase II to examine their abilities to enhance in situ biodegradation. In the first perturbation, nitrogen and phosphorus were spiked into the influent solution in excess of the required stoichiometric amounts. This perturbation did not have a significant impact because dispersion, not biokinetics, was the overall rate-limiting process. However, in the second perturbation, advection was increased, resulting in increased longitudinal and vertical transverse dispersion, thereby alleviating the rate-limiting process, and enhancing the overall biotransformation rate.

Sustainable oil and grease removal from synthetic stormwater runoff using bench-scale bioretention studies.

One of the principal components of the contaminant load in urban stormwater runoff is oil and grease (O&G) pollution, resulting from vehicle emissions. A mulch layer was used as a contaminant trap to remove O&G (dissolved and particulate-associated naphthalene, dissolved toluene, and dissolved motor oil hydrocarbons) from a synthetic runoff during a bench-scale infiltration study. Approximately 80 to 95% removal of all contaminants from synthetic runoff was found via sorption and filtration. Subsequently, approximately 90% of the sorbed naphthalene, toluene, oil, and particulate-associated naphthalene was biodegraded within approximately 3, 4, 8, and 2 days after the event, respectively, based on decreases in contaminant concentrations coupled with increases of microbial populations. These results indicate the effectiveness and sustainability of placing a thin layer of mulch on the surface of a bioretention facility for reducing O&G pollution from urban stormwater runoff.

Engineered bioretention for removal of nitrate from stormwater runoff.

A bioretention unit is a simple, plant- and soil-based, low-impact treatment and infiltration facility for treating stormwater runoff in developed areas. Nitrate, however, is not attenuated in conventional bioretention facilities. Thus, this study systematically evaluated a reengineered concept of bioretention for nitrate removal via microbial denitrification, which incorporates a continuously submerged anoxic zone with an overdrain. Experimental studies were performed in four phases. In the first two phases, column studies demonstrated that, overall, newspaper is the best solid-phase electron-donor substrate for denitrification out of the set studied (alfalfa, leaf mulch compost, newspaper, sawdust, wheat straw, wood chips, and elemental sulfur) based on superior nitrate removal and effluent water quality. The nitrate loading and hydraulic loading studies in the second phase provided design information. In the third phase, system viability after 30- and 84-day dormant periods was evaluated in column studies, demonstrating that newspaper-supported biological denitrification should be effective under conditions of intermittent loadings. Finally, in the fourth phase, pilot-scale bioretention studies demonstrated the effectiveness of the proposed design, showing nitrate plus nitrite mass removals of up to 80%. These results indicate that engineered bioretention for the removal of nitrogen from stormwater runoff has the potential for successful application as an urban stormwater treatment practice.

Bioenhancement of NAPL pool dissolution: experimental evaluation.

Experiments were conducted to quantify nonaqueous phase liquid (NAPL) pool dissolution and its enhancement by in situ biodegradation. The experiments were performed using square cross-section, glass-bead packed column reactors with a small pool of a toluene-in-dodecane mixture (toluene mole fraction, X(tol) approximately 0.02 or 0.09). Experimental quasi-steady-state toluene dissolution fluxes were determined using a 14C-carbon mass-balance approach during water flushing with and without biodegradation. The experiments demonstrated a statistically significant bioenhancement of the toluene dissolution flux of up to roughly twofold at average pore water velocities of approximately 0.1 and 1 m/day when the toluene mole fraction was low ( approximately 0.02); however, little or no bioenhancement was observed with the higher mole fraction ( approximately 0.09). Although it cannot be determined conclusively, the weight of evidence based on biomass measurements and model analyses suggests that the reduced bioenhancement for the high mole fraction was due to higher dissolved toluene concentrations, which may have caused toxicity effects. Importantly, even though NAPL dissolution was not bioenhanced in every case, the biodegradation reduced toluene concentrations to low levels in the reactor effluents.

Kinetics of phenol and chlorophenol utilization by Acinetobacter species.

Although microbial transformations via cometabolism have been widely observed, the few available kinetic models of cometabolism have not adequately addressed the case of inhibition from both the growth and nongrowth substrates. The present study investigated the degradation kinetics of self-inhibitory growth (phenol) and nongrowth (4-chlorophenol, 4-CP) substrates, present individually and in combination. Specifically, batch experiments were performed using an Acinetobacter isolate growing on phenol alone and with 4-CP present. In addition, batch experiments were also performed to evaluate the transformation of 4-CP by resting, phenol-induced Acinetobacter cultures. The Haldane kinetic model adequately predicted the biodegradation of phenol alone, although a slight discrepancy was noted in cases of higher initial phenol concentrations. Similarly, a Haldane model for substrate utilization was also able to describe the trends in 4-CP transformation by the resting cell cultures. The 4-CP transformation by the Acinetobacter species growing on phenol was modeled using a competitive kinetic model of cometabolism, which included growth and nongrowth substrate inhibition and cross-inhibition terms. Excellent agreement was obtained between the model predictions using experimentally estimated parameter values and the experimental data for the synchronous disappearance of phenol and 4-CP.