Chlorinated solvents such as trichloroethene (TCE) and perchloroethene (PCE) are commonly found in industrialized areas and can have major impact on human health and groundwater quality. The techniques for removing these substances from the subsurface environment is constantly being tuned and revised, and as such, the need for monitoring at such remediation sites is crucial. To find important correlations and hidden patterns between variables principle component analyses (PCA) and correlations matrixes were used on sets of field data from an existing remediation site in southern Sweden. Four important components were extracted in the following order; End products of dechlorination (EPD), second wave of dechlorination (SWD), first wave of dechlorination (FWD) and indicators of dechlorination (ID). The underlying pattern found in the data set was most likely derived from thermodynamic preference, explaining important correlations such as the correlation between iron and sulfate, the correlation between redox and degree of dechlorination. The law of thermodynamic preference means that we can (roughly) estimate the level of difficulty and/or the time it will take to remediate a polluted site.  These findings show that similar results shown in theory and laboratory environments also applies in the field and also that PCA is a potent tool for evaluating large data sets in this field of science. However, it is of great importance that the correlations are examined thoroughly, as correlation it not equal to causation.

This research investigates the physiological and transcriptional responses of Nitrosomonas europaea when exposed to chlorinated aromatic compounds and heavy metals under varying environmental conditions. It was found that transcriptional responses of identified sentinel genes correlate well with nitrification inhibition. Sorption of metals to biomass was also investigated and found not to correlate well with N. europaea inhibition. Whole genome microarray experiments were performed to define the transcriptional response of N. europaea when exposed to chlorobenzene. 13 out of 2460 N. europaea genes were significantly up-regulated after a 1-hour exposure to 4 μM chlorobenzene. HPLC analysis revealed that chlorobenzene was being oxidized primarily into 4-chlorophenol, and further physiological studies revealed that the presence of 4-chlorophenol could account for the inhibitory responses observed. RT-qPCR analysis of several differentially regulated genes verified that similar transcriptional responses were occurring for both chlorobenzene and 4-chlorophenol. 50% inhibitory concentrations of chlorobenzene and 4-chlorophenol resulted in moderate up-regulation of studied genes, however, increasing the concentration of 4-chlorophenol to achieve nitrification inhibition of 93% or more dramatically increased the fold regulation of several of the identified up- and down- regulated genes of interest. Increasing the 4-chlorophenol exposure time to 3 hours at the higher inhibition levels led to a general decrease in amplitude of transcriptional response for all genes tested. Cultures of N. europaea were exposed to various amounts of cadmium in aqueous solution containing EDTA, a strong metal-chelating organic, to control free ionic cadmium²⁺ (Cd²⁺) concentrations. Inhibition of ammonia oxidation as well as transcriptional up-regulation of merA, an identified sentinel gene for exposure to cadmium was found to correlate well with the concentration of Cd²⁺. The concentration of Cd²⁺ required to significantly affect N. europaea cells was found to be in the nanomolar range, which is several orders of magnitude lower than values reported in the literature for cadmium inhibition to mixed-culture activated sludge systems. The sorption of cadmium to the cells was found to be proportional to both the concentration of total cadmium and the concentration of Cd²⁺. At the concentration of metals required to cause approximately 50% nitrification inhibition, specific oxygen uptake results indicate the inhibition is specific to AMO with HAO and downstream energy-generation processes intact. To investigate more closely the inhibitory interactions between heavy metals and AMO, N. europaea inhibition by cadmium, zinc and silver was studied under substrate-limiting conditions. Unlike incubation in oxic environments, 1 hour incubations of N. europaea with cadmium and silver under anoxic conditions did not cause inhibition of nitrification activity after re-suspension in oxic media. In contrast, zinc, which is normally considered an analogue of cadmium in terms of toxic effect and transport mechanisms, was non-inhibitory to N. europaea when exposed in media lacking ammonia. Transcriptional response of merA closely followed the inhibition patterns, with samples which were inhibited after the removal of the metal having significant up-regulation of the gene, and those samples which were uninhibited showing no significant change in merA transcript levels compared to controls. Although sorbed metal concentrations were not found to be predictive of either extent of inhibition or transcriptional response, significantly more cadmium, zinc and silver were sorbed to biomass when incubated in aerobic media compared to anoxic media. Sorption in oxic media was found to be independent of AMO activity and similar results were obtained using Deinococcus radiodurans, a non-nitrifying gram-positive extremophile. The results indicate that greater heavy metal sorption to biomass in oxic environments may be due to general membrane chemistry effects. / Graduation date: 2011 / Access restricted to the OSU Community at author's request from March 31, 2011 - March 31, 2012

A study was carried out in a sedimentary fractured rock site located in south central Wisconsin, US, which was impacted by DNAPL releases estimated to occur in the 1950’s and 1960’s. The majority of the DNAPL has accumulated in the upper portion of the Lone Rock Formation at a depth between 140 and 180 ft bgs referred as Layer 5 in this study. A groundwater VOC plume of more than 3km long has formed in this Layer. The DNAPL is mainly composed of 1,1,1-TCA, PCE, TCE and BTEX, while large amounts of biodegradation products such as cis-DCE and 1,1-DCA are present in the plume. Long term VOC data have been collected at the site and diverse geological and hydrogeological techniques have been applied to have a better understanding of the DNAPL history and behavior of the VOC plume. Evidence of biodegradation was also documented near the DNAPL source in these studies. The thesis objectives of the present study aimed first to have a better understanding of the long term contaminant distribution and degradation history at the site. This objective was accomplished reviewing the VOC historical concentration data collected from 1992 to 2006 in the wells tapping the most contaminated. hydrogeological unit in the bedrock (Layer 5) and in the overburden aquifer (referred as Layer 2). The second objective aimed to evaluate the current degree or extent of biodegradation of chlorinated compounds, which was accomplished evaluating the current groundwater redox conditions and using a combined analysis of VOC concentration and carbon isotope data collected in groundwater in September 2007. The historical data collected between1992 to 2006 showed the degradation of the VOC plume in Layer 5 was controlled by the availability of electron acceptors and redox conditions in the fracture bedrock aquifer. This pattern and the extension of the VOC plume were linked to different DNALP pumping events in the source zone and the operation of a Hydraulic Barrier System. The current geochemical and isotope study showed a different pattern of biodegradation of chlorinated compounds in different parts of the plume. The cis-DCE tend to accumulate in the area from the source to the middle of the plume and around 80 % of biodegradation of 1,1,1-TCA to 1,1-DCA was observed in this area. The fringes of the plume were characterized by a dominant presence of TCE and 1,1,1 TCA. These patterns were linked to different redox conditions and amount of electron acceptors. The cis-DCE dominated area is characterized by anaerobic conditions and the presence of relative high amount of BTEX. The TCE-dominated area is under aerobic condition and no BTEX was found in this area. The operation of the Hydraulic Barrier System seems to have change redox condition which influenced the extent of degradation in the plume, especially in the area between the extraction wells. The formation of large amounts of VC in Layer 2 and the more reducing (at least sulfate reducing and maybe methanogenic conditions) of the groundwater in this Layer compared to Layer 5 confirmed the extent of VOC biodegradation is linked to the availability of electron donors. This study provides information about the current degree of the biodegradation of chlorinated compounds at a fracture rock site. This information is very valuable for the evaluation of natural attenuation as strategy for long term plume management or for future remediation strategies such as biostimulation or bioaugmentation at the site. This study also shows the present and long term behavior of the chlorinated compounds (degradation history) in the most contaminated hydrogeologic unit (Layer 5), has mainly been controlled by plume management strategies including DNAPL pumping in the source and the creation of a Hydraulic Barrier System. The ketones and BTEX, that acted as electron donors and carbon substrate for the microbial community responsible for the dechlorination of chlorinated compounds were shown to have controlled the past and current redox conditions and thus the degree and potential of biodegradation of chlorinated ethenes and chlorinated ethanes at the study site.

A study was carried out in a sedimentary fractured rock site located in south central Wisconsin, US, which was impacted by DNAPL releases estimated to occur in the 1950’s and 1960’s. The majority of the DNAPL has accumulated in the upper portion of the Lone Rock Formation at a depth between 140 and 180 ft bgs referred as Layer 5 in this study. A groundwater VOC plume of more than 3km long has formed in this Layer. The DNAPL is mainly composed of 1,1,1-TCA, PCE, TCE and BTEX, while large amounts of biodegradation products such as cis-DCE and 1,1-DCA are present in the plume. Long term VOC data have been collected at the site and diverse geological and hydrogeological techniques have been applied to have a better understanding of the DNAPL history and behavior of the VOC plume. Evidence of biodegradation was also documented near the DNAPL source in these studies. The thesis objectives of the present study aimed first to have a better understanding of the long term contaminant distribution and degradation history at the site. This objective was accomplished reviewing the VOC historical concentration data collected from 1992 to 2006 in the wells tapping the most contaminated. hydrogeological unit in the bedrock (Layer 5) and in the overburden aquifer (referred as Layer 2). The second objective aimed to evaluate the current degree or extent of biodegradation of chlorinated compounds, which was accomplished evaluating the current groundwater redox conditions and using a combined analysis of VOC concentration and carbon isotope data collected in groundwater in September 2007. The historical data collected between1992 to 2006 showed the degradation of the VOC plume in Layer 5 was controlled by the availability of electron acceptors and redox conditions in the fracture bedrock aquifer. This pattern and the extension of the VOC plume were linked to different DNALP pumping events in the source zone and the operation of a Hydraulic Barrier System. The current geochemical and isotope study showed a different pattern of biodegradation of chlorinated compounds in different parts of the plume. The cis-DCE tend to accumulate in the area from the source to the middle of the plume and around 80 % of biodegradation of 1,1,1-TCA to 1,1-DCA was observed in this area. The fringes of the plume were characterized by a dominant presence of TCE and 1,1,1 TCA. These patterns were linked to different redox conditions and amount of electron acceptors. The cis-DCE dominated area is characterized by anaerobic conditions and the presence of relative high amount of BTEX. The TCE-dominated area is under aerobic condition and no BTEX was found in this area. The operation of the Hydraulic Barrier System seems to have change redox condition which influenced the extent of degradation in the plume, especially in the area between the extraction wells. The formation of large amounts of VC in Layer 2 and the more reducing (at least sulfate reducing and maybe methanogenic conditions) of the groundwater in this Layer compared to Layer 5 confirmed the extent of VOC biodegradation is linked to the availability of electron donors. This study provides information about the current degree of the biodegradation of chlorinated compounds at a fracture rock site. This information is very valuable for the evaluation of natural attenuation as strategy for long term plume management or for future remediation strategies such as biostimulation or bioaugmentation at the site. This study also shows the present and long term behavior of the chlorinated compounds (degradation history) in the most contaminated hydrogeologic unit (Layer 5), has mainly been controlled by plume management strategies including DNAPL pumping in the source and the creation of a Hydraulic Barrier System. The ketones and BTEX, that acted as electron donors and carbon substrate for the microbial community responsible for the dechlorination of chlorinated compounds were shown to have controlled the past and current redox conditions and thus the degree and potential of biodegradation of chlorinated ethenes and chlorinated ethanes at the study site.

Polycyclic aromatic hydrocarbon (PAH) formation and growth from cyclopentadiene (CPD) moieties have been investigated using a laminar flow reactor and molecular modeling. The resonance-stabilized cyclopentadienyl radical is readily formed in flames and can participate in PAH growth to soot by reaction with the ??onds of aromatic species. Both CPD pyrolysis and computational results indicate that formation of indene and benzene is favored at low temperatures (below 750oC) and formation of naphthalene is favored at high temperatures. Reaction pathways from CPD have further been extended to PAH formation from the reaction of CPD and aromatic compounds with different types of ??onds. Results indicate that, while the major products from the pyrolysis of CPD, acenaphthylene, styrene and phenanthrene mixtures are from the reaction of CPD to itself rather than to these aromatic compounds with different ??onds, CPD does add to these compounds to produce larger PAH. Polychlorinated naphthalene (PCN) formation from chlorinated phenols has also been studied. In combustion exhaust gas, chlorinated phenols can produce dioxin as well as PCNs. PCN and polychlorinated dibenzofuran (PCDF) congener product distributions were consistent with proposed pathways involving phenoxy radical coupling at unchlorinated ortho-carbon sites. Tautomerization of the phenoxy radical coupling and subsequent fusion via H2O loss results in PCDF formation. Competing with this reaction pathway, CO elimination and subsequent fusion via hydrogen and/or chlorine loss was found to produce PCNs. PCDF isomer distributions were found to be weakly dependent to temperature, whereas PCN isomer distributions were found to be more temperature sensitive with selectivity to particular isomers decreasing with increasing temperature. Results of this research contribute to a better understanding of chemical mechanisms involved in the formation of toxic byproducts and soot in combustion systems.

In this study, a chlorinated-solvent contaminated groundwater site was used as the study site. Multivariate statistical analysis explains the huge and complicated current situation of the original data efficiently, concisely, and explicitly; it simplifies the original data into representative factors, or bases on the similarity between data to cluster and identify clustering outcome. The statistical software SPSS 12.0 was used to perform the multivariate statistical analysis to evaluate groundwater quality characteristics of this site. Results show that 20 analytical items of groundwater quality of the study site are simplified into seven major representative factors through factor analysis, including ¡§background¡¨, ¡§salt residual¡¨, ¡§hardness¡¨, ¡§ethylene chloride¡¨, ¡§alkalinity¡¨, ¡§organic pollutant¡¨, and ¡§chloroform¡¨. The factor score diagram was drawn according to the score of monitoring well on each factor and 89.6% of the variance could be obtained. This study used cluster analysis to cluster in two phrases, the groundwater quality monitoring wells were classified into seven clusters according to the similarity of monitored data nature and the differences between clusters. The groundwater quality characteristics and pollutant distributions of each cluster out this site were evaluated. The clustering result indicates that for the sixth cluster (where monitoring well SW-6 was the representative well), the average concentrations of chlorides such as 1,1-dichloroethylene, 1,1-dichloroethane, and cis-1,2-dichloroethylene were the highest among the clusters, indicating those the groundwater of nearby area might be polluted by chlorinated organic compounds. In addition, to evaluate whether the clustering of cluster analysis were appropriate or not, discriminant analysis is used to evaluate clustering accuracy, in which seven Fisher discriminant coefficient formulas that were exclusively suitable for this location were established. Then, the observed values were substituted to Fisher discriminant coefficient formula. Result shows that the monitoring well¡¦s clusters obtained from discriminant analysis were totally identical with the result of actual cluster analysis; the accuracy were 100%. After performing cross-validation analysis, the result shows that the accuracy were 80%, indicating the use of discriminant analysis (with forecasting function) to verify the clustering result of the cluster analysis was highly accurate. After analyzing the pollution condition of this site using time trend and space distribution, it were determined to conclude that trichloroethylene and 1,1-dichloroethylene were the major concerning pollutants; the pollutants appeared to be spreading on a large scale, so it was difficult to use the existing data to evaluate the pollution source. After assessing environmental medium characteristics and pollutant distribution of the site, this study suggests that the use of insitu bioremediation, which is cost-effective, can be applied as a remedial mothod.

Groundwater at many existing and former industrial sites and disposal areas is contaminated by halogenated organic compounds that were released into the environment. The chlorinated solvent trichloroethylene (TCE) is one of the most ubiquitous of these compounds. In situ chemical oxidation (ISCO) has been successfully used for the removal of TCE. The objective of this study was to apply the ISCO technology to remediate TCE-contaminated groundwater. In this study, potassium permanganate (KMnO4) was used as the oxidant during the ISCO process. The study consisted bench-scale and pilot-scale experiments. In the laboratory experiments, the major controlling factors included oxidant concentrations, effects of soil oxidant demand (SOD) on oxidation efficiency, and addition of dibasic sodium phosphate on the inhibition of production of manganese dioxide (MnO2). Results show that higher molar ratios of KMnO4 to TCE corresponded with higher TCE oxidation rate under the same initial TCE concentration condition. Moreover, higher TCE concentration corresponded with higher TCE oxidation rate under the same molar ratios of KMnO4 to TCE condition. Results reveal that KMnO4 is a more stable and dispersive oxidant, which is able to disperse into the soil materials and react with organic contaminants effectively. Significant amount of MnO2 production can be effectively inhibited with the addition of Na2HPO4. Results show that the increase in the first-order decay rate was observed when the oxidant concentration was increased, and the half-life was approximately 24.3 to 251 min. However, the opposite situation was observed when the second-order decay rate was used to describe the reaction. Results from the column experiment show that the breakthrough volumes were approximately 50.4 to 5.06 pore volume (PV). Injection of KMnO4 would cause the decrease in TCE concentration through oxidation. Results also indicate that the addition of Na2HPO4 would not inhibit the TCE removal rate. In the second part of this study, a TCE-contaminated site was selected for the conduction of pilot-scale study. A total of eight remediation wells were installed for this pilot-scale study. The initial TCE concentrations of the eight wells were as follows: C1 = 0.59 mg/L, C1-E = 0.64 mg/L, C1-W = 0.61 mg/L, EW-1 = 0.65 mg/L, EW-1E = 0.62 mg/L, EW-1W = 0.57 mg/L, C2 = 0.62 mg/L, C3 = 0.35 mg/L. C1, EW-1, C2, and C3 were located along the groundwater flow direction from the upgradient (C1) to the downgradient location (C3), and the distance between each well was 3 m. C1-E and C1-W were located in lateral to C1 with a distance of 3 m to C1. EW-1E and EW-1W were in lateral to EW-1 with a distance of 3 m to EW-1. In the first test, 2,700 L of KMnO4 solution was injected into each of the three injection wells (C1, C1-E, and C1-W) with concentration of 5,000 mg/L. Three injections were performed with an interval of 6 hr between each injection. After injection, the TCE concentrations in those three wells dropped down to below detection limit (<0.0025 mg/L). However, no significant variations in TCE concentrations were observed in other wells. In the second test, 2,700 L of KMnO4 solution was injected into injection well (EW-1) with concentration of 5,000 mg/L. Six injections were performed with an interval of 6 hr between each injection. After injection, the TCE concentrations in the injection well dropped down to below detection limit (<0.0025 mg/L). TCE concentrations in (C1, C1-E, C1-W, EW-1E, EW-1W, C2, and C3) dropped to 0.35-0.49 mg/L. After injection, no significant temperature and pH variation was observed. However, increase in conductivity and oxidation-reduction potential (ORP) was observed. This indicates that the KMnO4 oxidation process is a potential method for TCE-contaminate site remediation. The groundwater conductivity increased from 500 £gS/cm to 1,000 £gS/cm, and ORP increased from 200 to 600 mv. Increase in KMnO4, MnO2, and total Mn was also observed in wells. Results from the slug tests show that the hydraulic conductivity remained in the range from 10-4 to 10-5 m/sec before and after the KMnO4 injection.

Naturally occurring bacterial populations are capable of detoxifying chlorinated compounds and immobilizing the radionuclide uranium via reductive processes. This study addressed the following three knowledge gaps in the fields of chlorinated solvent and uranium bioremediation, 1) the risks and benefits of coupling bioremediation with thermal treatment for clean-up of chlorinated ethene-contaminated sites, 2) the accuracy of available techniques for the monitoring of chlorinated solvent bioremediation, and 3) the role of gram positive Desulfitobacterium spp. in uranium immobilization. Experiments demonstrated that thermal treatment increases electron donor availability, but the increased electron donor was not used to fuel reductive dechlorination and was actually consumed for methanogenesis. Two approaches for monitoring chlorinated solvent bioremediation were investigated, molecular techniques and compound-specific isotope analysis (CSIA). Results demonstrated that while Dehalococcoides (Dhc) gene expression was up-regulated under conditions inhibitory to dechlorination, the isotope effects associated with dechlorination reactions catalayzed by Dhc populations in consortia and in pure cultures were similar. U(VI) reduction by multiple Desulfitobacterium isolates was demonstrated. Interestingly, while almost all U(VI)-reducing populations have been reported to produce uraninite (UO2), the product of U(VI) reduction by Desulfitobacterium isolates was a unique form of insoluble mononuclear U(IV).

The two objectives of this work were to develop a simplified method for the analysis of chlorinated organics in water samples and to improve an existing soil remediation technology. The contaminants considered for these studies were chlorinated hydrocarbons because of their relative frequency of appearance at contaminated sites. The first half of this study involved the analysis of chlorinated ethenes by gas chromatography with flame ionization detection (GC-FID). I tested the hypothesis that the FID response factor is the same for all chlorinated ethene compounds. The rationale for this investigation is that if the hypothesis is correct, a single calibration curve can be used for GC/FID analysis of all chlorinated ethene compounds, saving time and money during sample analysis. Based on my measurements, a single calibration curve fits PCE, TCE, and cis-DCE (R2=0.998). However, the apparent slope of the calibration curve for vinyl chloride is approximately 45% lower, indicating that a separate calibration curve must be used to quantify vinyl chloride. I believe this difference in vinyl chloride is due to loss of analyte mass due to volatilization. The second half of the study considered the effect of solvent composition for a soil remediation technology, entitled remedial extraction and catalytic hydrodehalogenation (REACH), developed by Dr. Hun Young Wee and Dr. Jeff Cunningham (Wee and Cunningham, 2008). The objective of this thesis is to convert 1,2,4,5-tetrachlorobenzene (TeCB) to cyclohexane, thus improving on the work of Wee (2007). Recent work by Osborn (2011) tested successfully the use of palladium and rhodium catalysts for this conversion, though it took twelve hours for full conversion. Osborn (2011) performed her experiments in a 50:50 water-ethanol solvent; previous work by Wee and Cunningham (2008) suggests that using a 67:33 water-ethanol composition may dramatically reduce the reaction time. Therefore, the goal of this research was to use palladium and rhodium catalysts with a 67:33 water-ethanol solvent composition, with an aim of reducing the reaction time required to fully convert benzene to cyclohexane. The data suggest that the time required for conversion of the analyte to its product was improved dramatically compared to previous experiments. However, powdered palladium catalyst was used in this study instead of pellet form as in previous studies. The powdered palladium allowed for full conversion of the target chemical, TeCB, to benzene in less than 5 minutes. Benzene was fully converted to cyclohexane within 45 minutes in the batch reactor when a rhodium catalyst was used jointly with palladium. This study suggests that the 67:33 water-ethanol solvent composition be utilized in continuous flow tests in the future to improve the efficiency of the REACH system. The results also suggest that powdered palladium catalyst be considered because of its ability to force the reaction to completion in significantly less time than previous experiments.

Zero-valent metals such as bulk iron and zinc are known to dechlorinate toxicorganic compounds. Enhancement in reaction rates has been achieved through bimetallicnanosized particles such as nickel/iron (Ni/Fe) and palladium/iron (Pd/Fe). Batchdegradation of model compounds, trichlroethylene (TCE) and 2,2'-dichlorobiphenyls(DCB), were conducted using bimetallic Ni/Fe and Pd/Fe nanoparticles. Completedegradation of TCE and DCB is achieved at room temperature. Zero-valent iron, as themajor element, undergoes corrosion to provide hydrogen and electrons for the reductivecatalytic hydrodechlorination reaction. The second dopant metals of nickel and palladium(in nanoscale) act as catalyst for hydrogenation through metal hydride formation thatproduces completely dechlorinated final product. Different compositions of bimetallicNi/Fe and Pd/Fe nanoparticles were synthesized and their reactivity was characterized interms of reaction rate constants, hydrogen generation through iron corrosion, andproducts formation. The observed TCE degradation rate constant was two orders ofmagnitude higher than the bulk iron and nanoiron, indicating that the bimetallicnanoparticles are better materials compared to the monometallic iron systems. Longevitystudy through repeated cycle experiments showed minimum loss of activity. The surfacearea-normalized rate constant was found to have a strong correlation with the hydrogengeneration by iron corrosion reaction. A mathematical model was derived thatincorporates the reaction and Langmuirian-type sorption terms to estimate the intrinsicreaction rate constant and rate-limiting step in the degradation process. Bimetallicnanoparticles were also immobilized into the chitosan matrix for the synthesis of ananocomposite membrane reactor to achieve membrane-phase destruction of chlorinatedorganics under convective flow condition. Formation of uniformly distributed nanosizedparticles is confirmed by high resolution transmission electron microscopy. Themembrane-phase degradation results demonstrated similar trends with the previoussolution phase analysis with the observed enhanced reaction rates. The advantage of themembrane system is its ability to prevent the agglomeration of the nanoparticles in themembrane matrix, to minimize the loss of precious metals into the bulk solution phase,and to prevent the formation of precipitated Fe(III) hydroxide. These are due to thechelating effect of the amine and hydroxyl functional groups in the chitosan backbones.