A gas-fluidized bed bioreactor was developed in this research as a new method for treating polluted air. The fluidization characteristics of selected packing materials were investigated. Then, bioremediation was tested using two types of packing in a fluidized bioreactor, as well as in a comparable packed bed. Microorganisms on the particles biodegrade contaminants in the polluted air, which flows up through the bed. At high flowrates, the polluted air fluidizes the particles, while at low velocities the operation is in packed bed mode.<p>Initially, sawdust was selected for use as a packing material. Due to the poor fluidization properties of sawdust, glass spheres were added. A mixture of sawdust and glass spheres remained well mixed during fluidization. In the mixture, interparticle forces increased with increasing moisture in the sawdust, eventually causing defluidization of the bed. In the absence of bioremediation, mass transfer was studied between ethanol-contaminated air and sawdust/glass sphere packing, and found to be higher in the fluidized versus packed mode. In bioremediation experiments, ethanol removal efficiencies were as high as 95% in both operating modes. The maximum elimination capacities (EC) of ethanol were 75 and 225 g m^-3 sawdust h^-1 in the fluidized and packed beds respectively.<p>The packing of the fluidized bed bioreactor was optimized in order to boost bioremediation rates. Experiments showed that peat granules fluidized well in a bubbling regime, likely due to their relatively high density and sphericity. In peat bioremediation trials, the fluidized mode outperformed the packed bed; the maximum ECs were 1520 and 530 g m^-3 peat h^-1, respectively. Removal efficiency in the fluidized mode decreased with velocity, because the size and amount of large bubbles increased.<p>A steady-state model of the fluidized bioreactor was developed. By taking account of bubble properties during fluidization, the model helps to explain how bubble size, microbial properties and bioreactor residence time affect removal efficiency and elimination capacity of the bioreactor.<p>A peat gas-fluidized bioreactor shows promise as an efficient, low-cost technology for air treatment. Particle mixing in the fluidized bed may prevent operating problems associated with the packed bed bioreactor. Fluidized bioreactors are ideal for the treatment of high volume, low concentration air emissions.

The primary aim of the research summarized in this thesis was to confirm or refute that algae are involved in metal sedimentation from surface water, and whether this activity, if any, is enhanced by increased phosphorus availability. <p>A small-scale laboratory-based experiment was devised to elucidate the role of the chlorophyte alga Chlamydomonas noctigama in the removal of arsenic, nickel and uranium from mine water. Results indicated that the presence of <i>C. noctigama</i> significantly enhanced the removal of these metals relative to treatments without cells. Metals were present in greater concentrations in particulate matter in treatments with cells compared to treatments without cells, and there was a corresponding decrease in the concentrations of dissolved metals. This leads to the conclusion that sedimentation was mainly biotically induced. <p>Additional evidence of biotic involvement in metal removal from water by <i>C. noctigama</i> was obtained by using EDX spectroscopy and X-PEEM spectromicroscopy to observe complexation of arsenic, nickel and uranium to C. noctigama cells. Arsenic, the metal which was present at the lowest concentration in the DJX water, was present on scanned cells in low concentrations, and nickel and uranium, which were present at high concentrations in the DJX water, were present at higher concentrations. Examination of a single cell using X-PEEM spectromicroscopy showed uranium co-localized with carbon and phosphorus on the exterior of the cell. Crystalline particulate matter may have increased in the presence of cells. EDX spectroscopy showed that the crystalline particulate matter was possibly hydroxyapatite that contained various metals, including arsenic, nickel and uranium. EDX spectroscopy was used to determine the frequencies at which the cell-metal and particulate matter-metal associations occurred, and the relative concentrations of the metals associated with particulate matter. <p>No correlation between metal removal and phosphorus concentration in the media, or between algal density and phosphorus concentration was observed. However, phosphorus concentrations were not growth-limiting in these experiments, and so the effect of phosphorus on abiotic precipitation of metals remains unclear. <p> Results suggest two mechanisms by which <i>C. noctigama</i> may remove arsenic, nickel and uranium from solution: by direct sorption to the exterior of the cell, and by sorption to a cell product. <p>An experiment using cells preserved in Lugols iodine (a common phytoplankton sample preservative) indiated that Lugols preserved samples could not be used to quantify metals using spectroscopy. Consequently, historical samples preserved with Lugols iodine cannot be analyzed by this method.

University of Macau / Faculty of Science and Technology / Department of Civil and Environmental Engineering

University of Macau / Faculty of Science and Technology / Department of Civil and Environmental Engineering

Alternative strategies involving in situ remediation technologies have been developed to assist with property clean up, however, cost-effectiveness and discrepancies in success rates and timeliness continue. The objective of my research was to critically demonstrate the application and usefulness of an in situ remediation technology at a petroleum hydrocarbon impacted site. This project was proposed as part of the research programs: Groundwater Plume Formation and Remediation of Modern Gasoline Fuels in the Subsurface and Enhancing In Situ Bioremediation at Brownfield Sites funded by the Ontario Centres of Excellence for Earth and Environmental Technologies as part of the multiphase project entitled “Enhancing in situ Bioremediation at Brownfield Sites”. This research focused on the demonstration of nitrate-enhanced in situ bioremediation at a decommissioned service station. Petroleum hydrocarbon impacted soil and groundwater is a common occurrence at gasoline distribution facilities, where toxicological effects are known for gasoline constituents of interest such as benzene, toluene, ethylbenzene and total xylenes (BTEX). These chemicals are volatile, readily soluble, and persistent in groundwater. In particular, residual contaminants present in the saturated zone were targeted for remediation as they serve as a long term source of contamination and contribute to mobile vapour phase and dissolved phase plumes. Site investigations characterized the complex hydrogeological conditions and contaminant distribution present in order to effectively design an in situ bioremediation treatment system. The addition of nitrate as a terminal electron acceptor (TEA) to an aquifer enhances in situ biodegradation of petroleum hydrocarbons, by providing the microbes with a sustainable energy source to promote cell maintenance and growth of the microbial population. The remediation strategy involved pulsed injections of remedial solution amended with a conservative bromide (200 mg/L Br-) and reactive nitrate (90 to 265 mg/L NO3-) tracers with the purpose of providing a continuous supply of TEA available to the indigenous microbial populations. Nitrate was selected as an alternative electron acceptor over the thermodynamically favoured O2 because of typical challenges encountered using O2 in bioremediation applications in addition to the existing anaerobic environment. In situ anaerobic degradation of BTEX compound using TEA amendments has been well documented; however benzene is often recalcitrant under denitrification conditions. The results of the Br- tracer breakthrough curves indicate that different preferential flow pathways were established under the transient saturated conditions present at the Site, although the behaviour of the injected remedial slug was generally consistent between the different units and the test solution was ultimately delivered to the target zone. The delivery of the remedial test solution was greatly influenced by the hydrogeological conditions present at the time of injection. The injectate was preferentially transported in the high permeability zone of sandy gravel aquifer Unit 3 under high saturated condition and background hydraulic gradients. However the seasonal decline in groundwater levels and hydraulic gradients resulted in the lower portion of Unit 4 comprised of higher permeable materials being able to transmit the test solution more effectively. Given the variable hydrogeological conditions present at the Site influenced by seasonal effects, the delivery of the remedial solution to target zones containing petroleum hydrocarbons at residual saturation is more effective under reduced saturated conditions. The delivery of TEA amended water to enhance the in situ biodegradation of petroleum contaminants is more effective when the treatment water has an increased residence time in the target remedial zone, attributed to low gradients and groundwater transport velocities at the Site. Longer residence periods enable the indigenous microbes to have increased contact time with the TEA which will be preferentially utilized to degrade the contaminants.   A reducing zone enriched with TEA in the anaerobic aquifer was established following consecutive injections of remedial test solution. A cumulative mass of 4 kg of NO3- was added to the target aquifer during the course of the remedial injections. Evidence demonstrating NO3- utilized as a terminal electron acceptor in the bioremediation of the petroleum-contaminated aquifer include: laboratory microcosm study confirming local indigenous microbial population’s ability to degrade hydrocarbons using NO3- as the TEA in addition to observed decrease in NO3- relative to a conservative Br- tracer and generation of nitrite, an intermediate product in denitrification in the pilot-scale operation. Contaminant mass removal likely occurred as Br- tracer evidence indicates that NO3- was utilized in the study area based on the inference of denitrification rates. Post-injection groundwater sampling indicate declining concentrations of toluene, however long term monitoring is recommended in order to evaluate the success of the remediation activity and assess the potential for rebound. Post-injection soil core results are unable to demonstrate the reduction in individual toluene, let alone BTEXTMB hydrocarbon levels, as a result of insufficient quantities of nitrate delivered to the target zone relative to the significant but heterogeneously distributed residual mass in the subsurface.

A gas-fluidized bed bioreactor was developed in this research as a new method for treating polluted air. The fluidization characteristics of selected packing materials were investigated. Then, bioremediation was tested using two types of packing in a fluidized bioreactor, as well as in a comparable packed bed. Microorganisms on the particles biodegrade contaminants in the polluted air, which flows up through the bed. At high flowrates, the polluted air fluidizes the particles, while at low velocities the operation is in packed bed mode.<p>Initially, sawdust was selected for use as a packing material. Due to the poor fluidization properties of sawdust, glass spheres were added. A mixture of sawdust and glass spheres remained well mixed during fluidization. In the mixture, interparticle forces increased with increasing moisture in the sawdust, eventually causing defluidization of the bed. In the absence of bioremediation, mass transfer was studied between ethanol-contaminated air and sawdust/glass sphere packing, and found to be higher in the fluidized versus packed mode. In bioremediation experiments, ethanol removal efficiencies were as high as 95% in both operating modes. The maximum elimination capacities (EC) of ethanol were 75 and 225 g m^-3 sawdust h^-1 in the fluidized and packed beds respectively.<p>The packing of the fluidized bed bioreactor was optimized in order to boost bioremediation rates. Experiments showed that peat granules fluidized well in a bubbling regime, likely due to their relatively high density and sphericity. In peat bioremediation trials, the fluidized mode outperformed the packed bed; the maximum ECs were 1520 and 530 g m^-3 peat h^-1, respectively. Removal efficiency in the fluidized mode decreased with velocity, because the size and amount of large bubbles increased.<p>A steady-state model of the fluidized bioreactor was developed. By taking account of bubble properties during fluidization, the model helps to explain how bubble size, microbial properties and bioreactor residence time affect removal efficiency and elimination capacity of the bioreactor.<p>A peat gas-fluidized bioreactor shows promise as an efficient, low-cost technology for air treatment. Particle mixing in the fluidized bed may prevent operating problems associated with the packed bed bioreactor. Fluidized bioreactors are ideal for the treatment of high volume, low concentration air emissions.

The primary aim of the research summarized in this thesis was to confirm or refute that algae are involved in metal sedimentation from surface water, and whether this activity, if any, is enhanced by increased phosphorus availability. <p>A small-scale laboratory-based experiment was devised to elucidate the role of the chlorophyte alga Chlamydomonas noctigama in the removal of arsenic, nickel and uranium from mine water. Results indicated that the presence of <i>C. noctigama</i> significantly enhanced the removal of these metals relative to treatments without cells. Metals were present in greater concentrations in particulate matter in treatments with cells compared to treatments without cells, and there was a corresponding decrease in the concentrations of dissolved metals. This leads to the conclusion that sedimentation was mainly biotically induced. <p>Additional evidence of biotic involvement in metal removal from water by <i>C. noctigama</i> was obtained by using EDX spectroscopy and X-PEEM spectromicroscopy to observe complexation of arsenic, nickel and uranium to C. noctigama cells. Arsenic, the metal which was present at the lowest concentration in the DJX water, was present on scanned cells in low concentrations, and nickel and uranium, which were present at high concentrations in the DJX water, were present at higher concentrations. Examination of a single cell using X-PEEM spectromicroscopy showed uranium co-localized with carbon and phosphorus on the exterior of the cell. Crystalline particulate matter may have increased in the presence of cells. EDX spectroscopy showed that the crystalline particulate matter was possibly hydroxyapatite that contained various metals, including arsenic, nickel and uranium. EDX spectroscopy was used to determine the frequencies at which the cell-metal and particulate matter-metal associations occurred, and the relative concentrations of the metals associated with particulate matter. <p>No correlation between metal removal and phosphorus concentration in the media, or between algal density and phosphorus concentration was observed. However, phosphorus concentrations were not growth-limiting in these experiments, and so the effect of phosphorus on abiotic precipitation of metals remains unclear. <p> Results suggest two mechanisms by which <i>C. noctigama</i> may remove arsenic, nickel and uranium from solution: by direct sorption to the exterior of the cell, and by sorption to a cell product. <p>An experiment using cells preserved in Lugols iodine (a common phytoplankton sample preservative) indiated that Lugols preserved samples could not be used to quantify metals using spectroscopy. Consequently, historical samples preserved with Lugols iodine cannot be analyzed by this method.

Chlorinated aliphatic hydrocarbons (CAHs) include tetrachloroethene (PCE), trichloroethene (TCE), and others. The industrial solvent TCE is among the most ubiquitous chlorinated compounds found in groundwater pollution. TCE in environment can be removed by physical, chemical and biological procedures. Dehalorespiration is a biological pathway from which bacteria can derive energy from the reductive dechlorination of chlorinated ethenes using hydrogen or organic acids as electron donors and yielding chloride and ethene as degradation products. Dehalorespiration can be used to remediate chlorinated ethene contaminated aquifers if an appropriate aquifer ecosystem exists including populations of dechlorinating bacteria and companion organisms that contribute to the biogeochemical environment conducive to dehalorespiration activity. Enhanced in-situ aerobic or anaerobic bioremediation of chlorinated solvents is a cost-effective, expanding technology for the clean-up of chlorinated solvent-contaminated sites. The objective of this pilot-scale study was to apply an enhanced in situ bioremediation technology to remediate TCE-contaminated groundwater. Both aerobic and anaerobic remedial systems were evaluated at a TCE-spill site located in southern Taiwan. In the aerobic bioremediation zone, the effectiveness of air, nutrient, and sugarcane molasses injection to enhance the aerobic cometabolism on TCE degradation was evaluated. Results show that the decreases in TCE concentration were observed over 204 days operating period. Up to 73¢H-99¢H of TCE removal efficiency was obtained in this treatment system. In the anaerobic test zone, the effectiveness of nutrient and sugarcane molasses injection to enhance the anaerobic dechlorination on TCE degradation was also evaluated. Results show that the decreases in TCE concentration were observed over a 193-day operating period. Up to 53¢H-91¢H of TCE removal efficiency was obtained in this treatment system. Polymerase chain reaction was applied to analyze the gene variation in TCE-microbial degraders during the treatment process. Results from this study indicate that the aerobic TCE-degraders (type¢¹methanotrophs and type ¢º methanotrophs) and the gene of degradation enzymes (toluene monooxygenase, toluene dioxygenase, particulate methane monooxygenase) were detected after the treatment process in the aerobic test zone. In the anaerobic treatment zone, Dehalococcoides (anaerobic TCE-degrader) and the gene of degradation enzyme (vcrA and tceA) were detected and a significant drop of TCE concentration was also observed. Based on 16S rDNA sequence analysis, samples of groundwater from aerobic/anaerobic bioremediation zone are close related to the genera of Dehalococcoides sp. MB, Dehalococcoides ethenogenes 195, Dehalococcoides sp. VS, Acidovorax sp., Alicycliphilus sp., Burkholderiales, Caulobacter sp., Caulobacter tuntrae, Caulobacter vibrioides, Comamonadaceae, Hydrogenophaga sp., Iron-reducing bacterium, Mitsuaria chitosanitabida, Rhodocyclacea, Pseudomonas sp., Rhodoferax ferrireducens, Acinetobater sp., actinomycete, Pseudomonas aeruginosa and Variovorax sp. Results reveal that both the aerobic cometabolism and anaerobic dechlorination are feasible and applicable technologies to clean up TCE contaminated aquifers. Thus, the in situ bioremediation technology has the potential to be developed into an environmentally, economically and naturally acceptable remediation technology.

The goal of this study was to use molecular biology techniques to access and monitor the efficacy of bioremediation on a trichloroethene (TCE) polluted site. We added emulsified hydrogen releasing materials to stimulate onsite microbial growth and the biodegradation of TCE. This process was known as enhanced bioremediation. In this study, there were two bioremediation sites had been treated anaerobically. Groundwater samples were taken periodically for microbial analysis. Denaturing gradient gel electrophoresis (DGGE) was used to evaluate the variations in microbial community structures during the in situ groundwater remediation. The DGGE DNA bandings were sequenced to determine the 16S rRNA gene sequences and identify the dominate bacterial species. In addition, we used Dehalococcoides spp. 16S rRNA genes as the targets to do real-time PCR. Results show that the emulsified hydrogen releasing materials could enhance anaerobic reductive dechlorination. After addition of emulsified hydrogen releasing materials, we found that the volatile organic compounds concentrations (i.e., TCE, 1, 1-DCE and VC) were decreased. In microbial analysis, the diversities of the microbial community were increased after nutrient supplement. According to the DNA sequencing results, there were 31 bacterial species had been found that related to TCE degradation (i.e., Acidovorax sp., Burkholderiales, Pseudomonas sp., £]-proteobacterium, Comamonadaceae, Iron-reducing bacterium, Hydrogenophilaceae, Clostridium sp., Geobacter sp., Rhodoferax ferrireducens, Dehalospirillum multivorans and Dehalococcoides spp.). Dehalococcoides spp. can be used as a biomarker to evaluate the efficacy of anaerobic bioremediation on a TCE contaminated site. Therefore, we quantified Dehalococcoides populations to explain the capacity of bioremediation after addition of emulsified hydrogen releasing materials to groundwater. Results reveal that Dehalococcoides cell numbers of site A were 4.47¡Ñ103-8.26¡Ñ104 CFU/liter, site B were 4.60¡Ñ102-9.31¡Ñ107 CFU/liter. This data indicated that the addition of emulsified substrate would increase the growth of total Dehalococcoides population under anaerobic conditions. Overall, results from this study demonstrated that the microbial analysis and quantities of Dehalococcoides at different time points can provide useful information to proceed with bioremediation methods.

Soil and groundwater at many existing and former industrial areas and disposal sites is contaminated by organic solvent compounds that were released into the environment. Organic solvent compounds are heavier than water. When they are released into the subsurface, they tend to adsorb onto the soils and cause the appearance of LNAPL (light nonaqueous phase liquid) and DNAPL (dense nonaqueous phase liquid) pool. The industrial petroleum hydrocarbons (e.g., methyl tertiary-butyl ether, MTBE and benzene) and chlorinated solvent (e.g., trichloroethylene, TCE) are among the most ubiquitous organic compounds found in subsurface contaminated environment. One cost-effective approach for the remediation of the chlorinated solvent and petroleum products contaminated aquifers is the installation of permeable reactive zones or barriers within aquifers. As contaminated groundwater moves through the emplaced reactive zones, the contaminants are removed, and uncontaminated groundwater emerges from the downgradient side of the reactive zones. The objectives of this study were developed to evaluate the feasibility of applying in-situ chemical oxidation (ISCO) barrier and in-situ slow polycolloid-releasing substrate (SPRS) biobarrier system on the control of petroleum hydrocarbons and chlorinated solvent plume in aquifer. In the ISCO barrier system, it contained oxidant-releasing materials, to release oxidants (e.g., persulfate) contacting with water for oxidating contaminants existed in groundwater. In this study, laboratory-scale fill-and-draw experiments were conducted to determine the compositions ratios of the oxidant-releasing materials and evaluate the persulfate release rates. Results indicate that the average persulfate-releasing rate of 7.26 mg S2O82-/d/g was obtained when the mass ratio of sodium persulfate/cement/sand/water was 1/1.4/0.24/0.7. The column study was conducted to evaluate the efficiency of in situ application of the developed ISCO barrier system on MTBE and benzene oxidation. Results from the column study indicate that approximately 86-92% of MTBE and 95-99% of benzene could be removed during the early persulfate-releasing stage (before 48 pore volumes of groundwater pumping). The removal efficiencies for MTBE and benzene dropped to approximately 40-56% and 85-93%, respectively, during the latter part of the releasing period due to the decreased persulfate-releasing rate. Results reveal that acetone, byproduct of MTBE, was observed and then further oxidized completely. Results suggest that the addition of ferrous ion would activate the persulfate oxidation. However, excess ferrous ion would compete with organic contaminants for persulfate, causing the decrease in contaminant oxidation rates. In the SPRS biobarrier system, the food preparation industry has tremendous experiences in producing stable oil-in-water (W/O, 50/50) emulsions with a uniformly small droplet size. Surfactant mixture (71 mg/L of SL and 72 /L of SG) blending with water could yield a stable and the optimal emulsion was considered the best. The small absolute value of the emulsion zeta potential reduces inter-particle repulsion, causing the emulsion droplets to stick to each other when they collided. Overtime, large masses of flocculated droplets can form which then clog the sediment pores. The results can be used to predict abiotic interactions and distribution of contaminant mass expected after SPRS injection, and thus provides a more accurate estimate of the mass of TCE removed due to enhanced biodegradation. The effect of TCE partitioning to the vegetable oil on contaminant migration rates can be approximated using a retardation factor approach, where 0.28 years through a 3 m barrier. In anaerobic microcosm experiments, result show that SPRS can be fermented to hydrogen and acetate could be used as a substrate to simulate reductive dehalorination. The apparent complete removal of nitrate and sulfate by SPRS addition was likely a major factor that promoted the complete reduction of TCE at later stages of this study. Results from the column experiment indicate that occurrence of anaerobic reductive dechlorination in the biobarrier system can be verified by: (1) the oil: water partition coefficients of dissolved TCE into vegetable oil were be used to predict abiotic interactions and distribution of contaminant mass expected after SPRS injection. (2) The SPRS can ferment to hydrogen and acetate could be used as a substrate to simulate reductive dechlorination. The proposed treatment scheme would be expected to provide a more cost-effective alternative to remediate other petroleum hydrocarbons and chlorinated solvents-contaminated aquifers. Experiments and operational parameters obtained from this study provide an example to design a passive barriers system for in-site remediation.