Aerobic decomposition of hydrocarbon contaminants in anaerobic groundwater would be enhanced by oxygenating the water. This was done by injecting oxygen microbubbles in the soil matrix packed in a 7 ft by 7 ft by 5 inches in width Vertical Slice Test Cell, VSTC, and in a 30-inch column, also packed with sand. Transfer of oxygen to water was monitored after injecting oxygen microbubbles. Compared to sparged air and hydrogen peroxide injections documented in the literature to have transferred less than 2 percent oxygen to water, oxygen microbubbles transferred over 40 percent oxygen to the flowing groundwater. Also, after injection of microbubbles gas retentions over 70 percent were achieved. Oxygen Transfer Coefficients, KLa(s), were higher in layered soil in VSTC compared to non-layered soil when the same amounts of microbubbles were injected in the cell. The effect of cell layering, quality, stability, and the amount of microbubbles injections on transfer efficiency and gas holdup was studied. It was concluded that high initial gas holdups, KLa values oxygen transfer per time and percent oxygen transferred were important parameters in maintaining a sustained oxygen transfer zone. These experiments demonstrated that only one of these parameters can be at a maximum, say, a high percent oxygen transfer or a high percent initial retention or a high KLa value. However, a maximum value for one parameter is usually at the expense of the other two being low. The optimum values for these parameters would be dictated by the biochemical, sediment, and chemical oxygen demands placed on the oxygen transfer system. / Master of Science

by Chan Sze Sze. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 193-218). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstracts --- p.ii / 摘要 --- p.v / Contents --- p.viii / List of figures --- p.xv / List of tables --- p.xix / Abbreviations --- p.xxii / Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- The North Tsing Yi Abandoned Shipyard area --- p.1 / Chapter 1.2 --- Polycyclic aromatic hydrocarbons (PAHs) in the site --- p.3 / Chapter 1.2.1 --- Characteristics of PAHs --- p.3 / Chapter 1.2.2 --- Sources of PAHs --- p.8 / Chapter 1.2.3 --- Environmental fates of PAHs --- p.9 / Chapter 1.2.4 --- Biodegradation of PAHs --- p.10 / Chapter 1.2.5 --- Toxicity of PAHs --- p.13 / Chapter 1.2.6 --- PAHs contamination in Hong Kong --- p.14 / Chapter 1.2.7 --- Soil decontamination assessment in Hong Kong --- p.16 / Chapter 1.2.8 --- Environmental standards of PAHs --- p.18 / Chapter 1.2.9 --- Remediation technology of PAHs --- p.21 / Chapter 1.2.9.1 --- Bioremediation --- p.22 / Chapter 1.3 --- Heavy metals in the site --- p.28 / Chapter 1.3.1 --- "Characteristics of copper, lead and zinc" --- p.29 / Chapter 1.3.2 --- "Sources of copper, lead and zinc" --- p.32 / Chapter 1.3.3 --- "Environmental fates of copper, lead and zinc" --- p.34 / Chapter 1.3.4 --- "Toxicities of copper, lead and zinc" --- p.36 / Chapter 1.3.5 --- "Copper, lead and zinc contamination in Hong Kong" --- p.39 / Chapter 1.3.6 --- "Environmental standards of copper, lead and zinc" --- p.40 / Chapter 1.3.7 --- Remediation technology of heavy metal --- p.42 / Chapter 1.3.7.1 --- Chemical method --- p.42 / Chapter 1.3.7.2 --- Biological method --- p.43 / Chapter 1.3.7.3 --- Stabilization and Solidification --- p.45 / Chapter 1.4 --- Aim of study --- p.47 / Chapter 1.5 --- Objectives --- p.47 / Chapter 1.6 --- Research Strategy --- p.47 / Chapter 1.7 --- Significance of study --- p.48 / Chapter 2 --- Materials and Methods --- p.49 / Chapter 2.1 --- Soil Collection --- p.49 / Chapter 2.2 --- Characterization of soil --- p.49 / Chapter 2.2.1 --- Sample preparation --- p.49 / Chapter 2.2.2 --- "Soil pH, electrical conductivity & salinity" --- p.50 / Chapter 2.2.3 --- Total organic carbon contents --- p.51 / Chapter 2.2.4 --- Soil texture --- p.51 / Chapter 2.2.5 --- Moisture --- p.53 / Chapter 2.2.6 --- Total nitrogen and total phosphorus --- p.53 / Chapter 2.2.7 --- Available nitrogen --- p.53 / Chapter 2.2.8 --- Available phosphorus --- p.54 / Chapter 2.2.9 --- Soil bacterial and fungal population --- p.54 / Chapter 2.2.10 --- Extraction of PAHs and organic pollutants --- p.55 / Chapter 2.2.10.1 --- Extraction procedure --- p.55 / Chapter 2.2.10.2 --- GC-MS condition --- p.56 / Chapter 2.2.10.3 --- Preparation of mixed PAHs stock solution --- p.56 / Chapter 2.2.11 --- Oil and grease content --- p.57 / Chapter 2.2.12 --- Total Petroleum Hydrocarbons (TPH) --- p.57 / Chapter 2.2.13 --- Total heavy metal analysis --- p.58 / Chapter 2.2.14 --- Toxicity characteristic leaching procedure (TCLP) --- p.59 / Chapter 2.2.15 --- Extraction efficiency --- p.59 / Chapter 2.3 --- Production of mushroom compost --- p.60 / Chapter 2.4 --- Characterization of mushroom compost --- p.62 / Chapter 2.4.1 --- Enzyme assay --- p.62 / Chapter 2.4.1.1 --- Laccase assay --- p.62 / Chapter 2.4.1.2 --- Manganese peroxidase assay --- p.62 / Chapter 2.5 --- Addition of mushroom to soil on site --- p.63 / Chapter 2.5.1 --- Transportation of mushroom compost to Tsing Yi --- p.63 / Chapter 2.5.2 --- Mixing of mushroom compost and soil --- p.64 / Chapter 2.6 --- Soil Monitoring --- p.64 / Chapter 2.6.1 --- On site air and soil measurements --- p.64 / Chapter 2.6.1.1 --- Air temperature and moisture --- p.64 / Chapter 2.6.1.2 --- Light intensity --- p.64 / Chapter 2.6.1.3 --- UV intensity --- p.65 / Chapter 2.6.1.4 --- Rainfall --- p.65 / Chapter 2.6.1.5 --- Soil temperature --- p.65 / Chapter 2.6.2 --- Soil chemical characteristic --- p.65 / Chapter 2.6.3 --- Relative residue pollutant (%) --- p.65 / Chapter 2.7 --- Toxicity of treated soil --- p.66 / Chapter 2.7.1 --- Seed germination test --- p.66 / Chapter 2.7.2 --- Indigenous bacterial toxicity test --- p.67 / Chapter 2.7.3 --- Fungal toxicity test --- p.68 / Chapter 2.7.3.1 --- Preparation of ergosterol standard solution --- p.70 / Chapter 2.8 --- Soil Washing --- p.70 / Chapter 2.8.1 --- Optimization of soil washing --- p.70 / Chapter 2.8.1.1 --- Effect of hydrochloric acid concentration --- p.70 / Chapter 2.8.1.2 --- Effect of incubation time --- p.71 / Chapter 2.9 --- Phytoremediation --- p.71 / Chapter 2.10 --- Mycoextraction --- p.72 / Chapter 2.11 --- Integrated bioextraction --- p.72 / Chapter 2.12 --- Cementation --- p.73 / Chapter 2.13 --- Glass encapsulation --- p.73 / Chapter 2.14 --- Statistical analysis --- p.74 / Chapter 3 --- Results --- p.75 / Chapter 3.1 --- Characterization of soil --- p.75 / Chapter 3.2 --- Characterization of mushroom compost --- p.78 / Chapter 3.2.1 --- Enzyme activity --- p.78 / Chapter 3.2.2 --- Total nitrogen and total phosphorus contents --- p.78 / Chapter 3.3 --- Soil monitoring --- p.79 / Chapter 3.3.1 --- Initial pollutant content in biopile and fungal treatment soils --- p.79 / Chapter 3.3.2 --- On site air and soil physical characteristics --- p.81 / Chapter 3.3.2.1 --- Soil temperature and air temperature --- p.81 / Chapter 3.3.3 --- Soil chemical characteristic --- p.84 / Chapter 3.3.3.1 --- Effect of type of treatment on total petroleum hydrocarbon content --- p.85 / Chapter 3.3.3.2 --- Effect of type of treatment on oil and grease content --- p.87 / Chapter 3.3.3.3 --- Soil pH --- p.89 / Chapter 3.3.3.4 --- Moisture --- p.91 / Chapter 3.3.3.5 --- Electrical conductivity --- p.92 / Chapter 3.3.3.6 --- Salinity --- p.93 / Chapter 3.3.3.7 --- Microbial population --- p.95 / Chapter 3.3.3.8 --- Removal of organopollutant PAHs in biopile and fungal treatment --- p.98 / Chapter 3.3.3.9 --- Effect of type of treatment on residual PAHs at Day 4 --- p.104 / Chapter 3.3.3.10 --- Effect of type of treatment on residual PAHs at peak levels --- p.107 / Chapter 3.3.3.11 --- Effect of type of treatment on residual organopollutants at the end of treatments --- p.109 / Chapter 3.3.3.12 --- Effect of type of treatment on total nitrogen and phosphorus contents --- p.111 / Chapter 3.3.3.13 --- Effect of type of treatment on physical and chemical properties of soil --- p.113 / Chapter 3.4 --- Toxicity of treated soil --- p.116 / Chapter 3.4.1 --- Seed germination test --- p.116 / Chapter 3.4.2 --- Indigenous bacterial toxicity test --- p.120 / Chapter 3.4.3 --- Fungal toxicity test --- p.125 / Chapter 3.5 --- Soil washing --- p.129 / Chapter 3.5.1 --- Optimisation of soil washing --- p.129 / Chapter 3.5.1.1 --- The effect of hydrochloric acid concentration --- p.129 / Chapter 3.5.1.2 --- The effect of incubation time --- p.134 / Chapter 3.6 --- Mycoextraction --- p.139 / Chapter 3.7 --- Phytoextraction and integrated bioextraction --- p.146 / Chapter 3.8 --- Cementation --- p.153 / Chapter 3.9 --- Glass encapsulation --- p.158 / Chapter 4 --- Discussion --- p.160 / Chapter 4.1 --- Characterization of soil --- p.160 / Chapter 4.2 --- Characterization of mushroom compost --- p.162 / Chapter 4.2.1 --- Enzyme activity --- p.162 / Chapter 4.2.2 --- Total nitrogen and total phosphorus contents --- p.163 / Chapter 4.3 --- Soil monitoring --- p.163 / Chapter 4.3.1 --- Initial pollutant content in biopile and fungal treatment soil --- p.163 / Chapter 4.3.2 --- On site air and soil physical characteristics --- p.164 / Chapter 4.3.3 --- Soil chemical characteristic --- p.164 / Chapter 4.3.3.1 --- Soil pH --- p.164 / Chapter 4.3.3.2 --- Moisture --- p.165 / Chapter 4.3.3.3 --- Electrical conductivity --- p.165 / Chapter 4.3.3.4 --- Salinity --- p.166 / Chapter 4.3.3.5 --- Microbial population in biopile and fungal treatments --- p.166 / Chapter 4.3.3.6 --- Removal of organopollutant PAHs in biopile and fungal treatments --- p.168 / Chapter 4.3.3.7 --- Effect of type of treatment on residual PAHs at peak levels --- p.170 / Chapter 4.3.3.8 --- Effect of type of treatment on residual oil and grease and TPH contents --- p.171 / Chapter 4.3.3.9 --- Effect of type of treatment on total nitrogen and phosphorus contents --- p.172 / Chapter 4.3.3.10 --- Effect of type of treatment on physical and chemical properties of the soil --- p.173 / Chapter 4.4 --- Toxicity of treated soil --- p.174 / Chapter 4.5 --- Summary of Pleurotus pulmonarius mushroom compost on organopollutant remediation --- p.177 / Chapter 4.6 --- Soil washing --- p.178 / Chapter 4.7 --- Mycoextraction --- p.180 / Chapter 4.8 --- Phytoextraction and integrated bioextraction --- p.182 / Chapter 4.9 --- Cementation --- p.184 / Chapter 4.10 --- Glass encapsulation --- p.185 / Chapter 4.11 --- "Summary of physical, chemical and biological heavy metal removal treatments" --- p.186 / Chapter 4.12 --- Future studies --- p.187 / Chapter 5 --- Conclusion --- p.190 / Chapter 6 --- References --- p.193

An investigation of the biodegradation of complex hydrocarbon wastes, with emphasis on chlorinated aromatic compounds, in an anaerobic/aerobic bioprocess environment was made. A reactor configuration was developed consisting of linked anaerobic and aerobic reactors which served as the model for a proposed bioremediation strategy targeting subterranean soil/sediment/aquifer chlorinated phenol-contaminated environments. Here oxygen is frequently limited and sulphate is readily available, as occurs especially in marine sediment and intertidal habitats. In the anaerobic system the successful transformation and mobilization of the model contaminant, 2,4,6-trichlorophenol, was shown to rely on reductive dechlorination by a sulphate-reducing dependent dechlororespiring co-culture. This was followed in the aerobic system by degradation of the pollutant and its metabolites, 2,4-dichlorophenol, 4-chlorophenol and phenol, by immobilized white-rot fungi.The strategy was initially investigated separately in laboratory bench- and intermediate scale reactors whereafter reactors were linked to simulate the integrated biodegradation strategy. The application of the fungal reactor to treat an actual waste stream by degrading complex mixtures of hydrocarbons in a waste oil recycling effluent was also investigated. The mineralization of phenol and 2,4,6-TCP by immobilized fungal cultures was studied in pinewood chip and foam glass bead-packed trickling reactors. The reactors were operated in sequencing batch format. Removal efficiency increased over time and elevated influent phenol and TCP (800 and 85 mg.L⁻¹) concentrations were degraded by > 98 % in 24 – 30 h batch cycles. Comparable performance between the packing materials was shown. Uptake by the packing was negligible and stripping of compounds induced by aeration had a minimal effect on biodegradation efficiency. Reactor performances are discussed in relation to sequencing batch operation and nutrient requirements necessary to sustain fungal activity in inert vs. organic material packed systems. It was shown that a co-culture consisting of sulphate-reducing and dechlororespiring bacteria established in fed-batch and soil flasks, as well as pine chip-packed fluidized bed reactors. Results showed reductive dechlorination of 2,4,6-TCP to be in strict dependence on the activity of the sulphate-reducing population, sulphate and lactate concentrations. Transformation to 2,4-DCP, 4-CP and phenol was enhanced in sulphate deficient conditions. Dechlororespiring activity was found to be dependent on the fermentative activity of sulphate-reducing bacteria, and the culture was also shown to mobilize and dechlorinate TCP in soils contaminated with the pollutant. Linking the systems achieved degradation of the compound by > 99 % through fungal mineralization of metabolites produced in the dechlororespiring stage of the system. pH correction to the anaerobic reactor was found to be necessary since acidic effluent from the fungal reactor inhibited sulphate reduction and dechlorination. The fungal reactor system was evaluated at intermediate-scale using a complex waste oil recycling effluent. Substantial COD reduction (> 96 % in 48 h batch cycles) and removal of specific effluent hydrocarbon components was shown in diluted, undiluted (COD > 37 g.L⁻¹) and 2,4,6-TCP-spiked effluents. Industrial application of the fungal reactor was evaluated in a 14 m³ pilot plant operated on-site at a waste oil processing plant.

The overall goal of this work was to investigate microbial activity leading to the anaerobic degradation of polycyclic aromatic hydrocarbons and an organophilic clay sediment cap used at a creosote-contaminated Superfund site. To determine whether or not PAHs were being degraded under anaerobic conditions in situ, groundwater and sediment porewater samples were analyzed for metabolic biomarkers, or metabolites, formed in the anaerobic degradation of naphthalene (a low-molecular weight PAH). In addition, a groundwater push-pull method was developed to evaluate whether the transformation of deuterated naphthalene to a deuterated metabolite could be monitored in situ and if conservative rates of transformation can be defined using this method. Metabolites of anaerobic naphthalene degradation were detected in all samples that also contained significant levels of naphthalene. Anaerobic degradation of naphthalene appears to be widespread in the upland contaminated aquifer, as well as within the adjacent river sediments. A zero-order rate of transformation of naphthalene-D₈ to naphthoic acid-D₇was calculated as 31 nM·d-¹. This study is the first reported use of deuterated naphthalene to provide both conclusive evidence of the in situ production of breakdown metabolites and an in situ rate of transformation. Methane ebullition was observed in areas of the sediment cap footprint associated with organophilic clay that was used a reactive capping material to sequester mobile non-aqueous phase liquid (NAPL) at the site. Anaerobic slurry incubations were constructed using sediment core samples to quantify the contribution of the native sediment and the different layers of capping material (sand and organophilic clay) to the overall methane production. Substrate addition experiments using fresh, unused organophilic clay, as well as measured changes in total carbon in organophilic clay over time supported the hypothesis that microbes can use organophilic clay as a carbon source. Quantitative PCR (qPCR) directed at the mcrA gene enumerated methanogens in field samples and incubations of native sediment and capping materials. Denaturing gradient gel electrophoresis (DGGE) was also performed on DNA extracted from these samples to identify some of the predominant microorganisms within the sediment cap footprint. The organophilic clay incubations produced up to 1500 times more methane than the native sediment and sand cap incubations. The organophilic clay field sample contained the greatest number of methanogens and the native sediment contained the least. However, the native sediment incubations had greater numbers of methanogens compared to their respective field sample and comparable numbers to the organophilic clay incubation. An increase in methane production was observed with the addition of fresh, unused organophilic clay to the already active organophilic clay incubations indicating that organophilic clay stimulates methanogenesis. In addition, organophilic clay retrieved from the field lost about 10% of its total carbon over a 300-day incubation period suggesting that some component of organophilic clay may be converted to methane. DGGE results revealed that some of the predominant groups within the native sediment and sediment cap were Bacteriodetes, Firmicutes, Chloroflexi, and Deltaproteobacteria. An organism 98% similar to Syntrophus sp. was identified in the organophilic clay suggesting this organism may be working in concert with methanogens to convert the organic component of organophilic clay ultimately to methane. The capacity of organophilic clay to sequester organic contaminants will likely change over time as the organic component is removed from the clay. This, in turn, affects the use of this material as a long-term remedial strategy in reduced, contaminated environments.

The bioremediation of petroleum contaminated soil in large-scale treatment units was studied in conjunction with Sybron Chemicals Inc., Salem, VA. The soil had been previously contaminated and was spiked with additional petroleum. Water with different characteristics was circulated through the soil in order to evaluate the effect of nutrient enhanced treatment without and with addition of two inoculation materials - Sybron’s ABR Hydrocarbon Degraders and Rhodococcus sp. - on the rate of hydrocarbon degradation. Treatment units without nutrients and introduced organisms served as controls. Total petroleum hydrocarbon concentrations (TPH) were monitored using two alternative analytical methods, infrared spectrophotometry and gas chromatography. The results of the field study and different laboratory experiments, a radiotracer flask assay, static soil microcosms, and soil columns were compared. While nutrient addition did enhance biodegradation, the addition of autochthonous organisms was not found to accelerate hydrocarbon degradation rates in the previously contaminated soil. A significant decline of surface tension in the circulated water after inoculation with Rhodococcus, was thought to be due to microbial production of surfactants, but did not increase TPH degradation. The radiotracer technique and microcosm study confirmed these results. The soil column study indicated that the rapid degradation of soluble and slower degradation of less soluble hydrocarbons occurred in two subsequent phases with approximately zero order rates. Typical degradation rates for the more soluble or degradable petroleum hydrocarbons were approximately 40 ppm/week and for the less soluble and degradable compounds 10 ppm/week. Microcosms were found to successfully predict the degradation rates of the soluble hydrocarbons, while the soil columns simulated degradation of the less soluble hydrocarbons best. The analysis of soil extracts for petroleum hydrocarbon concentrations with infrared spectrophotometry was found to be defective. / Master of Science

The increased use of alcohols as gasoline additives, and possible substitutes, has prompted the investigation of the fate of gasoline/alcohol mixtures in the environment. In situ bioremediation is one technique that can successfully be applied to remove ground water contaminants particularly in situations where the adsorptive capacity of the soil plays a major role. Frequently, enhanced in situ bioremediation techniques rely on indigenous microorganisms to degrade ground water contaminants; this technique may sometimes include the addition of acclimated bacteria. In this study, soil microcosms were constructed in order to simulate the conditions found in a saturated aerobic aquifer. The biodegradation potential of methanol, benzene, and m-xylene was investigated. Uncontaminated soil from the surface, 12, 16.5, and 18 foot depths was utilized to observe the differences in microbial responses throughout the soil profile. The biodegradation potential of the indigenous microbiota was determined and compared to that of benzene acclimated bacteria, for all the compounds in the mixture. To observe the impact that chemical and physical soil characteristics may have on microbial responses, soils from each depth were classified on the basis of their particle size, moisture content and pH. Substantial methanol, benzene, and m-xylene biodegradation by the indigenous microorganisms occurred in all subsurface soils. While methanol was readily biodegradable over concentrations ranging from about 80 mg/L to about 200 mg/L, benzene inhibited methanol biodegradation at about 125 mg/L in all soil depths. The addition of benzene acclimated bacteria considerably increased the biodegradation rates of all compounds in the mixture. Such increases in biodegradation rates may be attributed to the activities of both groups, the indigenous microorganisms and the benzene acclimated bacteria. The results obtained by this study suggest that biodegradation of methanol, benzene, and m-xylene can readily occur in a saturated aerobic subsurface environment. The physical and chemical properties of a ground water aquifer seem to have a marked effect on microbial responses, and consequently on the biodegradation potential of water contaminants. / Master of Science

Two weathered, petroleum contaminated soils were studied to determine if weathered products are amenable to bioremediation and to determine which TPH (total petroleum hydrocarbon) fractions were degrading during particular time frames under different remediation alternatives. Delineation of fractional degradation patterns results in inferences regarding the efficacy of different treatment methods on various petroleum products. A sandy loam and a clay soil were both studied to determine if soil matrix affects the degradation patterns. The experimental matrix included sacrificial static microcosms, soil columns and aerated slurry reactors. Both soils were evaluated under all bioreactor configurations using both a nutrient amended water and a water lacking nutrients. Controls were also used to evaluate abiotic losses. Biodegradation rates generally followed a biphasic pattern, initially rapid then followed by a slow or stagnant period. Degradation rates increased from static microcosms to soil columns to slurry reactors. The slow phase was controlled by the presence of recalcitrant compounds which decreased in number and concentrations from static microcosms to columns to slurry reactors, and generally with nutrient addition. Nutrient addition enhanced degradation for all sandy soil treatments, but only slurry reactor treatment for the clay soil. The entire TPH spectrum was broken down into five minute parcels based on GC elution time. The compounds that eluted quickest generally were the easiest to degrade. The fraction that effectively covered the TPH components in gasoline was well removed under all treatment modes. All nutrient amended studies resulted in rapid essentially complete removal of the light fractions within two weeks. The fraction encompassing the middle distillates such as diesel fuel and jet fuel was degraded under all treatment methods, however only the slurry reactors resulted in final TPH levels that would have met regulatory limits. Fractions that eluted after 15 minutes were not effectively degraded by the static microcosms or the soil columns for either soil, eliminating in situ bioremediation as a viable treatment alternative for crude oil, fuel oil and gas oil contamination, at concentrations present in this study. Persistence of recalcitrant compounds was the major factor leading to the poor biodegradation observed in the static microcosms and soil columns. Fractional degradation was highly dependent on the initial concentration of the fraction. Generally, fractions present in the largest concentrations degraded fastest. / Master of Science

Thesis (MTech (Chemistry))--Cape Peninsula University of Technology, 2016. / Polycyclic aromatic hydrocarbons (PAHs) are an alarming group of organic substances for humans and environmental organisms due to their ubiquitous presence, toxicity, and carcinogenicity. They are semi-volatile substances which result from the fusion of carbon and hydrogen atoms and constitute a large group of compounds containing two to several aromatic rings in their molecule. Natural processes and several anthropogenic activities involving complete or incomplete combustion of organic substances such as coal, fossil fuel, tobacco and other thermal processes, generally result in the release of the PAHs into the environment. However, the fate of the PAHs is of great environmental concern due to their tendency to accumulate and their persistence in different environmental matrices and their toxicity. Animal studies have revealed that an excessive exposure to PAHs can be harmful. Evidence of their carcinogenic, mutagenic, and immune-suppressive effects has been reported in the literature. In the soil environment, they have the tendency to be absorbed by plants grown on soil being contaminated by the PAHs. It is, therefore, important to evaluate their occurrence levels in different environmental matrices such as soil concentrations.

Yip, Ho-yin. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 181-201). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstract --- p.ii / Contents --- p.vi / List of Figures --- p.x / List of Tables --- p.xvii / Abbreviations --- p.xix / Chapter 1. --- Introduction --- p.1 / Chapter 1.1 --- Polycyclic aromatic hydrocarbons (PAHs) --- p.1 / Chapter 1.1.1 --- Characteristics of PAHs --- p.1 / Chapter 1.1.2 --- Sources of PAHs --- p.2 / Chapter 1.1.3 --- Environmental fates of PAHs --- p.3 / Chapter 1.1.4 --- Effects of PAHs on living organisms --- p.5 / Chapter 1.1.4.1 --- General effects --- p.5 / Chapter 1.1.4.2 --- Effects on plants --- p.6 / Chapter 1.1.4.3 --- Effects on invertebrates --- p.7 / Chapter 1.1.4.4 --- Effects on fishes --- p.7 / Chapter 1.1.4.5 --- Effects on reptiles and amphibians --- p.8 / Chapter 1.1.4.6 --- Effects on birds --- p.9 / Chapter 1.1.4.7 --- Effects on mammals --- p.9 / Chapter 1.2 --- PAH contamination in Hong Kong --- p.10 / Chapter 1.3 --- Treatments of PAH contamination --- p.12 / Chapter 1.3.1 --- Physical treatments --- p.12 / Chapter 1.3.2 --- Chemical treatments --- p.13 / Chapter 1.3.3 --- Biological treatments --- p.14 / Chapter 1.4 --- Advanced oxidation processes (AOPs) --- p.16 / Chapter 1.5 --- Summary --- p.24 / Chapter 2. --- Objectives --- p.27 / Chapter 3. --- Materials and Methods --- p.28 / Chapter 3.1 --- Chemicals --- p.28 / Chapter 3.2 --- Photocatalytic reactor --- p.30 / Chapter 3.3 --- Determination of PAHs concentrations --- p.30 / Chapter 3.3.1 --- Extraction of PAHs --- p.30 / Chapter 3.3.2 --- Quantification of PAHs --- p.32 / Chapter 3.4 --- Optimization of physico-chemical conditions for PCO --- p.37 / Chapter 3.4.1 --- Determination of the reaction time for optimization of PCO --- p.37 / Chapter 3.4.2 --- Effect of titanium dioxide (Ti02) concentration and light intensity --- p.38 / Chapter 3.4.3 --- Effect of initial pH and hydrogen peroxide (H2O2) concentration --- p.38 / Chapter 3.4.4 --- Effect of initial PAHs concentration --- p.39 / Chapter 3.5 --- Toxicity analysis --- p.39 / Chapter 3.5.1 --- Microtox® test for acute toxicity --- p.39 / Chapter 3.5.2 --- Mutatox® test for genotoxicity --- p.42 / Chapter 3.6 --- Determination of total organic carbon (TOC) removal in optimized PCO --- p.43 / Chapter 3.7 --- Determination of degradation pathways --- p.43 / Chapter 3.7.1 --- Extraction of intermediates and/or degradation products --- p.45 / Chapter 3.7.2 --- Identification of intermediates and/or degradation products --- p.45 / Chapter 4. --- Results --- p.49 / Chapter 4.1 --- Determination of PAHs concentrations --- p.49 / Chapter 4.2 --- Optimization of extraction method --- p.49 / Chapter 4.3 --- Optimization of physico-chemical conditions for PCO --- p.49 / Chapter 4.3.1 --- Determination of the reaction time for optimization of PCO --- p.49 / Chapter 4.3.2 --- Effect of Ti02 concentration and light intensity --- p.60 / Chapter 4.3.3 --- Effect of initial pH --- p.88 / Chapter 4.3.4 --- Effect of initial H2O2 concentration --- p.99 / Chapter 4.3.5 --- Effect of initial PAHs concentration --- p.104 / Chapter 4.3.6 --- Improvements on removal efficiency (RE) after optimization --- p.113 / Chapter 4.4 --- Toxicity analysis --- p.122 / Chapter 4.4.1 --- Microtox® test for acute toxicity --- p.122 / Chapter 4.4.2 --- Mutatox® test for genotoxicity --- p.122 / Chapter 4.5 --- Determination of TOC removal in optimized PCO --- p.129 / Chapter 4.6 --- Determination of degradation pathways --- p.129 / Chapter 5. --- Discussion --- p.150 / Chapter 5.1 --- Determination of PAHs concentrations --- p.150 / Chapter 5.2 --- Optimization of extraction method --- p.150 / Chapter 5.3 --- Optimization of physico-chemical conditions for PCO --- p.151 / Chapter 5.3.1 --- Determination of the reaction time for optimization of PCO --- p.151 / Chapter 5.3.2 --- Effects of Ti02 concentration and light intensity --- p.152 / Chapter 5.3.3 --- Effects of initial pH --- p.160 / Chapter 5.3.4 --- Effects of initial H202 concentration --- p.163 / Chapter 5.3.5 --- Effects of initial PAHs concentration --- p.165 / Chapter 5.3.6 --- Improvements on RE after optimization --- p.167 / Chapter 5.4 --- Toxicity analysis --- p.169 / Chapter 5.4.1 --- Microtox® test for acute toxicity --- p.169 / Chapter 5.4.2 --- Mutatox® test for genotoxicity --- p.170 / Chapter 5.5 --- Determination of TOC removal in optimized PCO --- p.171 / Chapter 5.6 --- Determination of detoxification pathways --- p.172 / Chapter 6. --- Conclusion --- p.177 / Chapter 7. --- References --- p.181 / Chapter 8. --- Appendix I --- p.202

This study evaluated the efficacy of bioremediation as a treatment option for a hydrocarbon and heavy metal contaminated soil. Microbial degradation of hydrocarbons under aerobic, nitrate-reducing and sulphate-reducing conditions was examined. Nutrient supplementation with nitrogen and phosphate as well as aeration seemed to be the most important factors for enhancing biodegradation. From initial batch studies, a carbon: nitrogen ratio of 50: 1 was found to be optimal for biodegradation. However, very low carbon to nitrogen ratios were undesirable since these inhibited microbial activity. Manipulation of the pH did not seem to be beneficial with regard to hydrocarbon biodegradation. However, low pH values induced elevated concentrations of leachate heavy metals. Aerobic conditions provided optimal conditions for hydrocarbon catabolism with up to 54% of the original contaminant degraded after 2 months of treatment. Further treatment for up to 20 months did not significantly increase hydrocarbon biodegradation. Under nitrate- and sulphatereducing conditions, 6% and 31 % respectively of the initial contaminant was degraded after 2 months while after a further 20 months, 50% and 42%, respectively were degraded. The addition of soil bulking agents and the use of sparging did not significantly increase biodegradation. Similarly, the addition of inoculum did not influence biodegradation rates to any great degree. The presence of heavy metals up to concentrations of 400 mgt1 Mn, 176 mgt1 Zn and 94 mgt1 Ni did not reduce microbial activity within the soil. During the treatment phase, heavy metal and hydrocarbon migration were limited even under water saturation and low pH conditions. A Biodegradation Index was developed and evaluated and may, potentially, find use as an in situ assessment technique for microbial hydrocarbon catabolism. The iodonitrophenyltetrazolium salt assay was also found to be an effective and rapid alternative assay for monitoring bioremediation progress. / Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 1996.