In this field-scale study, air sparging (AS) system was applied at a petroleum-hydrocarbon spill site to remediate contaminated soil and groundwater in situ. The objective of this study was to evaluate the effectiveness of the AS system on volatile organic compounds (VOC) removal via the volatilization mechanism. Moreover, the AS system would also enhance the in situ bioremediation process due to the increased oxygen concentration in the subsurface. Results from the preliminary site characterization show that high concentrations of benzene and toluene were present in the subsurface in the western part of the site. Up to 15.62 and 30,957 mg/Kg of benzene and toluene were detected in soil samples, respectively. Moreover, up to 0.068 and 4.8 mg/L of benzene and toluene were observed in groundwater samples, respectively. The following remediation activities were conducted during the one-year investigation and remediation period: 1. Construction of four recovery wells were for light non-aqueous phase liquid (LNAPL) and contaminated groundwater extraction to prevent the expansion of VOC plume. The extracted groundwater was delivered to the wastewater treatment plant for treatment before discharge. 2. Installation of ten air sparging wells to enhance the removal of VOC through volatilization and biodegradation processes. 3. Conduction of (1) soil gas survey, (2) soil and groundwater sampling and analyses, and microbial enumeration periodically to evaluate the effectiveness of AS on VOC removal. Results from the field-scale study indicate that the AS system is able to effectively contain the plume. This can be confirmed by the following findings: (1) decrease in VOC concentrations in both soil and groundwater, (2) increase in carbon dioxide and increase in oxygen concentrations in the soil gas samples, and (3) increase in bacterial population in soil samples. Results from this study indicate that AS system can effectively contain the plume and manage this petroleum hydrocarbon spill site.

Abstract This research was to evaluate the treatment efficiency for in-situ treatment of pentachlorophenol (PCP) contaminated soil by electrokinetics-Fenton process combined with biodegradation. An electric gradient of 1V/cm, and graphite electrodes were employed in all experiments. Soil types, catalyst types and dosage, hydrogen peroxide concentration, cathode reservoir liquid species and reaction time were employed as the experimental factors in this study. In this study, no matter electrokinetics-Fenton process or the electrokinetics-biodegradation in the latter, prolong the reaction time can promote the removal and destruction efficiency (DRE) of target pollutant from soil. By using 0.0196 M FeSO4 with 3.5% H2O2, the DRE was only lower 2% than 0.098 M FeSO4 with 3.5% H2O2.It showed that using 0.0196 M FeSO4 can provide enough Fe2+ to react with H2O2. By increasing H2O2 concentration from 0.35% to 3.5%, a DRE rised from 68.34% to 79.77%. When iron powder was used as catalyst, the residual pentachloroplenol concentration near to anode reservoir lower than 0.0196 M FeSO4 was used. But the DRE was 56.58% lower than the 68.34% of using 0.0196 M FeSO4.As the influences of soil types to electrokinetics-Fenton process, the residual concentration of pollutant for Soil No. 2 was higher than Soil No. 1. A DRE of only 59.22% was obtained. It is postulated that a much higher content of organic matter with Soil No. 2 whereas lower the treatment efficiency because of consumption of hydroxyl radicals by the organic matter of soil. For the influence of different reservoir liquid species, in this study 0.1M acetic buffer solution was used as cathode reservoir liquid, expected to promote the removal efficiency. From the result of experiment that could not reach the expected treatment efficiency of increasing the removal efficiency from soil. From the experiment of electrokinetics process combined with cometabolism, a treatment efficiency of only 25.67% was obtained. The content of pollutant within every section of soil column were still higher than predict. But by using electrokinetics-Fenton process to pretreat the pollutant within soil first, the increasing efficiency of biodegradation was found. Even when reaction time was prolonged, the pollutant could be completely eliminated from soil. If only used iron minerals to proceed electrokinetics-Fenton process naturally exited in the soil, a DRE of only 20

This research was initiated after a 100-year flood caused an oil spill on the San Jacinto River (Houston, Texas) in October of 1994. After the floodwaters subsided the released petroleum floating on the water was deposited on the surrounding lands. The petroleum spill was used as an opportunity to research intrinsic petroleum biodegradation in a 9-acre petroleum impacted estuarine wetland. The first phase of this research (Phase I) began in December 1994, approximately 1.5 months after the spill of opportunity and involved the study and quantification of in-situ petroleum biodegradation. The second phase of the research (Phase II) began in March 1996 with a controlled oil release to study and evaluate the success of two bioremediation treatments versus natural biodegradation. The study of in-situ petroleum hydrocarbon degradation and the evaluation of bioremediation amendments were successfully quantified using GC-MS analytical techniques. However, the GC-MS technique is limited to the analyses of hydrocarbon compounds, a disadvantage that precludes the overall characterization of petroleum degradation. The research presented here details an analytical technique that was used to provide a full characterization of temporal petroleum biodegradation. This technique uses thin layer chromatography coupled with flame ionization detection (TLC-FID) to characterize the saturate and aromatic (hydrocarbon) fractions and the resin and asphaltene (non-hydrocarbon, polar) fractions. Other analysis techniques, such as HPLC-SARA analysis, are available for the full characterization of the four petroleum fractions. However, these techniques do not lend themselves well to the application of large sample set analysis. A significant advantage of the TLC-FID analysis to other petroleum analysis techniques is the ability to analyze several samples concurrently and quickly with relative ease and few resources. For the purposes of the Phase I and Phase II research the TLC-FID analysis method was evaluated, refined and applied to quantify the temporal biodegradation and bioremediation of petroleum. While the TLC-FID analysis produces a full characterization, it cannot supplant the GC-MS analysis for petroleum bioremediation research. However, it can be used in conjunction with the GC-MS to expand the knowledge of petroleum bioremediation and remediation strategies.

Toxic waste disposal problems have become enormous due to the proliferation of xenobiotic compounds for use in agricultural, industrial and numerous other applications. Organophosphate (OP) pesticides are commonly used in agriculture and their toxicity is associated with inhibition of cholinesterase in the exposed organism. Some OPs have been shown to produce OP-induced delayed neuropathy (OPIDN). The overall goal of the work described in this thesis was to develop bacterial consortia to remediate hazardous substances at significantly higher rates than found with natural systems. Specifically, degradation of methyl parathion (MP) by hydrolysis with a genetically engineered Escherichia coli was investigated along with degradation of one of the resulting products, p-nitrophenol (PNP), by Sphingobium chlorophenolicum ATCC 53874. Simultaneous degradation of both MP and PNP was investigated using a consortium of a genetically engineered Escherichia coli and a native S. chlorophenolicum. Concentrations of MP and PNP were measured by high performance liquid chromatography (HPLC). Non-growing freely suspended recombinant OPH+ E. coli cells efficiently degraded MP without addition of nutrients for growth. Maximum reactor productivity was found with a biomass concentration of 25 g/L. Substrate inhibition did not occur up to 3 g MP/L. The simple Michaelis-Menten kinetic model for enzymatic reactions provided a good fit of the degradation data with Vm=11.45 ??mol/min??g-biomass and Km=2.73 g/L. B. cepacia failed to degrade PNP under the experimental conditions evaluated, so further studies were not conducted. Growing cultures of S. chlorophenolicum degraded PNP at concentrations up to 0.1 g/L without a lag phase in mineral salts glutamate medium. Parameters such as initial pH, growth medium and growth stage for addition of PNP were important degradation factors. The bacterium exhibited substantial growth in the degradation process. Hydroquinone (HQ) or nitrocatechol (NC) were not identified as products of PNP degradation. The recombinant OPH+ E. coli and S. chlorophenolicum consortium failed to degrade PNP when starting with higher concentrations of MP. The presence of organic solvent in the bacterial consortium degradation medium negatively affected the degradation of PNP. The genetically engineered organism efficiently degraded high concentrations of MP, but the resulting high concentration of intermediate product (PNP) inhibited growth of the native type organism. Biodegradation by consortia of genetically engineered non-growing and native-type organisms generally will be limited by the growing native-type organism.

The biodegradability and physicochemical properties of unsubstituted and methylated polycyclic aromatic hydrocarbons (PAHs) were investigated. The focus was on the development of models expressing the influence of molecular structure and properties on observed behavior. Linear free energy relationships (LFERs) were developed for the estimation of aqueous solubilities, octanol/water partition coefficients, and vapor pressures as functions of chromatographic retention time. LFERs were tested in the estimation of physicochemical properties for twenty methylated naphthalenes containing up to four methyl substituents. It was determined that LFERs can accurately estimate physicochemical properties for methylated naphthalenes. Twenty unsubstituted and methylated PAHs containing up to four aromatic rings were biodegraded individually by Sphingomonas paucimobilis strain EPA505, and Monod-type kinetic coefficients were estimated for each PAH using the integral method. Estimated extant kinetic parameters included the maximal specific biodegradation rate, the affinity coefficient, and the inhibition coefficient. The generic Andrews model adequately simulated kinetic data. The ability of PAHs to serve as sole energy and carbon sources was also evaluated. Quantitative structure-biodegradability relationships (QSBRs) were developed based on the estimates of the kinetic and growth parameters. A genetic algorithm was used for QSBR development. Statistical analysis and validation demonstrated the predictive value of the QSBRs. Spatial and topological molecular descriptors were essential in explaining biodegradability. Mechanistic interpretation of the kinetic data and the QSBRs provided evidence that simple or facilitated diffusion through the cell membranes is the rate-determining step in PAH biodegradation by strain EPA505. A kinetic experiment was conducted to investigate biodegradation of PAH mixtures by strain EPA505. The investigation focused on 2-methylphenanthrene, fluoranthene, and pyrene, and their mixtures. Integrated material balance equations describing different interaction types were fitted to the depletion data and evaluated on a statistical and probabilistic basis. Mixture degradation was most adequately described by a pure competitive interaction model with mutual substrate exclusivity, a fully predictive model utilizing parameters estimated in the sole-PAH experiments only. The models developed in this research provide insight into how molecular structure and properties influence physicochemical properties and biodegradability of PAHs. The models have considerable predictive value and could reduce the need for laboratory testing.

Natural nitro compounds with diverse structures and biological functions are produced by bacteria, fungi, plants and animals. Little is known about the behavior of such compounds in natural ecosystems. The lack of accumulation in the biosphere implies that they are biodegraded. Microbial strategies for biodegradation of synthetic nitro compounds are well established; however only two pathways are known for degradation of natural nitro compounds. The research described here examines the genes that encode the key enzymes required for biodegradation of 3-nitrotyrosine (3NTyr) and 1-nitro-2-phenylethane (NPE). 3NTyr is a biological marker for disease and inflammation in plants and animals. A 3NTyr degrading microbe, Variovorax sp. JS669 was isolated from soil. We identified the JS669 denA, which encodes an enzyme that catalyzes denitration of 4-hydroxy-3-nitro-phenylacetate, the key step in metabolism of 3NTyr. The isolation of 3NTyr degraders and development of molecular probes specific to denA revealed that 3NTyr degradation is a widespread phenomena in natural habitats and the compound is metabolized by phylogenetically diverse bacteria. Phylogenetic analysis of the 4-hydroxy-3-nitro-phenylacetate denitrase from JS669 revealed it to be the first functionally annotated protein in a clade of unidentified Class A flavin monooxygenases. NPE has been identified from several plants, yet the biodegradation of the compound remained a mystery. Here we report the degradation of NPE and its analog 2-nitropropylbenzene. Discovery of the metabolic pathway revealed a novel microbial strategy to use a meta-ring fission degradation pathway to cleave an undesirable side chain from an aromatic compound and use the remainder of the compound as a carbon and energy source. Two genes that encode enzymes in the biodegradation pathway were identified and both are deeply branched within their respective phylogenetic trees, indicating that both represent highly specialized microbial enzymes. Furthermore, microbial degradation of NPE resulted in the production of 3-nitropropionic acid, a natural toxin that inhibits succinate dehydrogenase and is responsible for livestock illness and death. This is the first report of bacterial production of 3-nitropropionic acid, and might represent a significant source of 3-nitropropionic acid in natural habitats. The findings from these studies contribute to the overall understanding of microbial metabolism. Specifically, this research reveals genes that encode novel enzymes and strategies for the biodegradation of two natural nitro compounds. Furthermore, discovery of mechanisms for the biodegradation of such compounds reveals novel microbial metabolic diversity and provides insight into the evolution of degradation pathways for synthetic compounds.

Thesis (Ph.D)--Earth and Atmospheric Sciences, Georgia Institute of Technology, 2010. / Committee Chair: Taillefert, Martial; Committee Member: DiChristina, Thomas; Committee Member: Sobecky, Patricia; Committee Member: Van Cappellen, Philippe; Committee Member: Webb, Samuel. Part of the SMARTech Electronic Thesis and Dissertation Collection.

Capping is typically used to control contaminant release from the underlying sediments. However, the presence of conventional caps often eliminates or slows natural degradation that might otherwise occur at the surface sediment. This is primarily due to the development of reducing conditions within the sediment that discourage hydrocarbon degradation. The objective of this study was to develop a novel active capping method, an electrode enhanced cap, to manipulate the redox potential to produce conditions more favorable for hydrocarbon degradation and evaluate the approach for the remediation of PAH contaminated sediment. A preliminary study of electrode enhanced biodegradation of PAH in sediment slurries showed that naphthalene and phenanthrene concentration decreased significantly within 4 days, and PAH degrading genes increased by almost 2 orders of magnitude. In a sediment microcosm more representative of expected field conditions, graphite cloth was used to form an anode at the sediment-cap interface and a similar cathode was placed a few centimeters above within a thin sand layer. With the application of 2V voltage, ORP increased and pH dropped around the anode reflecting water electrolysis. Various cap amendments (buffers) were employed to moderate pH changes. Bicarbonate was found to be the most effective in laboratory experiments but a slower dissolving buffer, e.g. siderite, may be more effective under field conditions. Phenanthrene concentration was found to decrease slowly with time in the vicinity of the anode. In the sediment at 0-1 cm below the anode, phenanthrene concentrations decreased to ~70% of initial concentration with no bicarbonate, and to ~50% with bicarbonate over ~70 days, whereas those in the control remained relatively constant. PAH degrading gene increased compared with control, providing microbial evidence of PAH biodegradation. A voltage-current relationship, which incorporated separation distance and the area of the electrodes, was established to predict current. A coupled reactive transport model was developed to simulate pH profiles and model results showed that pH is neutralized at the anode with upflowing groundwater seepage. This study demonstrated that electrode enhanced capping can be used to control redox potential in a sediment cap, provide microbial electron acceptors, and stimulate PAH degradation. / text

published_or_final_version / Ecology and Biodiversity / Doctoral / Doctor of Philosophy

The study examined the removal mechanism of non-acclimated and acclimated aerobic activated sludge for 29 target organic micropollutants (OMPs) at low concentration. The selection of the target OMPs represents a wide range of physical-chemical properties such as hydrophobicity, charge state as well as a diverse range of classes, including pharmaceuticals, personal care products and household chemicals. The removal mechanisms of OMPs include adsorption, biodegradation, hydrolysis, and vaporization. Adsorption and biodegradation were found to be the main routes for OMPs removal for all target OMPs. Target OMPs responded to the two dominant removal routes in different ways: (1) complete adsorption, (2) strong biodegradation and weak adsorption, (3) medium biodegradation and adsorption, and (4) weak sorption and weak biodegradatio. Kinetic study showed that adsorption of atenolol, mathylparaben and propylparaben well followed first-order model (R2: 0.939 to 0.999) with the rate constants ranging from 0.519-7.092 h-1. For biodegradation kinetics, it was found that benzafibrate, bisphenol A, diclofenac, gemfibrozil, ibuprofen, caffeine and DEET followed zero-order model (K0:1.15E-4 to 0.0142 μg/Lh-1, R2: 0.991 to 0.999), while TCEP, naproxen, dipehydramine, oxybenzone and sulfamethoxazole followed first-order model (K1:1.96E-4 to 0.101 h-1, R2: 0.912 to 0.996). 4 Inhibition by sodium azide (NaN3)and high temperature sterilization was compared, and it was found that high temperature sterilization will damage cells and change the sludge charge state. For the OMPs adaptation removal study, it was found that some of OMPs effluent concentration decreased, which may be due to the slow adaptation of the sludge or the increase of certain bacteria culture; some increased due to chromic toxicity of the chemicals; most of the OMPs had stable effluent concentration trend, it was explained that some of the OMPs were too difficutl to remove while other showed strong quick adaptation. A new module combined of sequencing batch reactor (SBR) and nanofiltration membrane filtration (NF-MBR) was developed to further study the OMPs removal and to exam the concept of compounds (CRT). The NF-MBR was proved to be a promising bioreactor, as OMPs were rejected by NF membrane which leaded to a low OMPs concentration in permeate water, the apparent removal rate was over 80% for most of the OMPs.