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The uptake, distribution and depuration of a water soluble, mono-aromatic hydrocarbon contained in petroleum and refined products was studied in two species of marine fish, Nature northern anchovy (Engraulis mordax) and juvenile striped bass (Horone saxatilis) were exposed to sub-lethal concentrations of 14c benzene for 48 hours. Residues in tissues exhibiting a high lipid content (non-metabolic pathway) or representing apparent major metabolic sites were measured during the exposure and afterwards when the fish were transferred to clean seawater. Fish exhibited a rapid uptake over a wide range of benzene concentrations in the water column. Accumulation in anchovy was considerably greater than in striped bass. Results indicate that the pathway of hydrocarbons through the liver, gall bladder, intestines and colon is a major depuration route. Residues were depurated rapidly after cessation of exposure; in striped bass tissues. most residues were undetectable by seven days,

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

A laboratory scale study was conducted to investigate the treatability of petroleum contaminated soils by soil washing and subsequent biological treatment of the different soil fractions. In addition to soils obtained from contaminated sites, studies were also performed on soils contaminated in the laboratory. Soil washing was performed using a bench-scale soil washing system. Washing was carried out with simultaneous fractionation of the bulk soil into sand, silt and clay fractions. Cleaning efficiencies due to soil washing ranged from 60 to 81% for different soils. After washing, the finer soil particles (silts and clays) were found to possess higher concentrations of adsorbed hydrocarbons, thus requiring further treatment. Distribution of n-alkanes was studied to obtain a better understanding of contaminant redistribution and mass transfer during washing. / Master of Science

The purpose of this study was to compare the simultaneous effects of the water soluble fraction (WSF) of JP-4, a common military and civilian jet fuel, on survival, ventilatory rate, preference-avoidance behavior, and blood enzymes and ions of the bluegill sunfish (Lepomis rnacrochirus) to determine possible interrelationships and which procedures might be most descriptive of sublethal stress. The dynamic 96-h LCSO for WSF JP-4 was 26.2% (percent of the maximum soluble amount of JP-4). A concentration of 5.1% WSF JP-4 caused a detectable shift in ventilatory functions (rate and amplitude); this concentration was equivalent to the 96-h LCOl. Fish did not display a strong preference-avoidance reaction when exposed to WSF JP-4. Some avoidance occurred at 3.5% and 4.9% WSF JP-4. At concentrations near the 96-h LCSO fish appeared to lose their ability to detect WSF JP-4, indicating potential for fish not to avoid lethal levels of WSF JP-4. Few significant changes in whole and serum blood parameters were measured in fish exposed to 13% WSF JP-4; at near lethal concentrations changes were primarily attributed to osmoregulatory failure. Therefore such changes in blood parameters would not be useful to validate or confirm exposure to WSF JP-4. There were few significant changes in gill and liver histology of fish exposed to WSF JP-4 other than an increase in size or number of lipid vacuoles in the liver of exposed fish. / Ph. D.

Lai Ho-yan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1999. / Includes bibliographical references (leaves 104-122). / Abstracts in English and Chinese. / ACKNOWLEDGEMENT --- p.I / ABSTRACT (ENGLISH) --- p.II / ABSTRACT (CHINESE) --- p.IV / TABLE OF CONTENT --- p.VI / LIST OF FIGURES --- p.IX / LIST OF TABLES --- p.V / INTRODUCTION --- p.1 / Chapter 1.1 --- Background --- p.1 / Chapter 1.2 --- Sources of oil pollution --- p.1 / Chapter 1.3 --- Composition of oil --- p.4 / Chapter 1.4 --- Fate of oil in the environment --- p.6 / Chapter 1.5 --- Toxic effect of oil on marine ecosystem --- p.8 / Chapter 1.5.1 --- Acute toxicity --- p.8 / Chapter 1.5.2 --- Chronic toxicity --- p.9 / Chapter 1.5.3 --- Carcinogenicity of oil --- p.11 / Chapter 1.6 --- The origins of ecotoxicology --- p.12 / Chapter 1.7 --- Need for ecotoxicity tests --- p.13 / Chapter 1.8 --- Testings in ecotoxicology --- p.15 / OBJECTIVES --- p.17 / MATERIALS AND METHODS --- p.18 / Chapter 1. --- Collection of oily wastes samples --- p.18 / Chapter 2. --- Preparation of samples --- p.18 / Chapter 2.1 --- Water-soluble fraction --- p.19 / Chapter 2.2 --- Polycyclic aromatic hydrocarbon fraction --- p.20 / Chapter 2.2.1 --- Supercritical fluid extraction --- p.20 / Chapter 2.2.2 --- Silica gel chromatography --- p.22 / Chapter 2.2.3 --- Sample concentration --- p.22 / Chapter 3. --- Chemical analyses of oily wastes --- p.26 / Chapter 3.1 --- Determination of heavy metal concentration in oily waste samples --- p.26 / Chapter 3.1.1 --- Nitric acid-perchloric acid digestion --- p.26 / Chapter 3.1.2 --- Inductively coupled plasma-emission spectrometric analysis --- p.26 / Chapter 3.2 --- Determination of polycyclic aromatic hydrocarbon concentration in oily waste samples --- p.28 / Chapter 3.2.1 --- Determination of polycyclic aromatic hydrocarbon concentration in water- soluble fraction --- p.28 / Chapter a. --- Liquid-liquid extraction --- p.28 / Chapter b. --- Gas chromatography-mass spectrometric analysis of water-soluble fraction --- p.29 / Chapter 3.2.2 --- Determination of polycyclic aromatic hydrocarbon concentration in crude oily waste samples --- p.32 / Chapter a. --- Supercritical fluid extraction and silica gel column chromatography --- p.32 / Chapter b. --- Gas chromatography-mass spectrometric analysis of polycyclic aromatic hydrocarbon fraction --- p.33 / Chapter 4 --- ecotoxicological studies of oily wastes --- p.34 / Chapter 4.1 --- Toxicity tests and sample preparation --- p.34 / Chapter 4.2 --- Ecotoxicological studies of water soluble fraction --- p.34 / Chapter 4.2.1 --- "Growth inhibition test on a marine alga, Chlorella pyrenoidosa CU-2" --- p.34 / Chapter 4.2.2 --- "Survival test on a marine amphipod, Elasmopus rapax" --- p.39 / Chapter 4.2.3 --- "Survival test on a marine fish, Ambassis gymnocephalus" --- p.41 / Chapter 4.2.4 --- Microtox® test --- p.43 / Chapter 4.3. --- Ecotoxicological studies of polycyclic aromatic hydrocarbon fraction --- p.45 / Chapter 4.3.1 --- "Growth inhibition test on a marine alga, Chlorella pyrenoidosa CU-2" --- p.45 / Chapter 4.3.2 --- "Survival test on a amphipod, Parhyale plumulosa" --- p.45 / Chapter 4.3.3 --- "Survival test on the fish, Sparus sarba" --- p.47 / Chapter 4.3.4 --- Microtox® test --- p.49 / Chapter 5. --- Statistical analyses of chemical and ecotoxicological analyses --- p.50 / RESULTS --- p.51 / Chapter 1. --- Chemical analyses of oily wastes --- p.51 / Chapter 1.1 --- Inductively coupled plasma-emission spectrometric analysis --- p.51 / Chapter 1.1.1 --- Heavy metal concentration in crude oily wastes --- p.51 / Chapter 1.1.2 --- Heavy metal concentration in water-soluble fraction --- p.51 / Chapter 1.1.3 --- Heavy metal concentration in ploy cyclic aromatic hydrocarbon fraction --- p.54 / Chapter 1.2 --- Gas chromatography- mass spectrometry analysis --- p.54 / Chapter 1.2.1 --- Polycyclic aromatic hydrocarbon concentration in crude oily wastes --- p.54 / Chapter 1.2.2 --- Polycyclic aromatic hydrocarbons concentration in water-soluble fraction --- p.59 / Chapter 1.2.3 --- Polycyclic aromatic hydrocarbons concentration in polycyclic aromatic hydrocarbon fraction --- p.61 / Chapter 2. --- Ecotoxicological studies of oily wastes --- p.63 / Chapter 2.1 --- Ecotoxicological studies of water-souble fraction --- p.63 / Chapter 2.1.1 --- Growth inhibition test on Chlorella pyrenoidosa CU-2 --- p.63 / Chapter 2.1.2 --- Survival test on Elasmopous rapax --- p.63 / Chapter 2.1.3 --- Survival test on Ambassis gymnocephalus --- p.67 / Chapter 2.1.4 --- Microtox® test --- p.67 / Chapter 2.2 --- Ecotoxicological studies of polycyclic aromatic hydrocarbon fraction --- p.70 / Chapter 2.2.1 --- Growth inhibition test on Chlorella pyrenoidosa CU-2 --- p.70 / Chapter 2.2.2 --- Survival test on Parhyale plumulosa --- p.70 / Chapter 2.2.3 --- Survival test on Sparus sarba --- p.74 / Chapter 2.2.4 --- Microtox® test --- p.74 / Chapter 3. --- Statistical analyses of chemical and ecotoxicological studies --- p.77 / Chapter 3.1 --- Statistical analyses of studies on water-soluble fraction --- p.77 / Chapter 3.1.1 --- Correlation between heavy metal concentration in water-soluble fraction and toxicity tests --- p.77 / Chapter 3.1.2 --- Correlation between concentration of total polycyclic aromatic hydrocarbon in water-soluble fraction and toxicity tests --- p.80 / Chapter 3.1.3 --- Correlation among acute toxicity tests --- p.80 / Chapter 3.2 --- Statistical analyses of polycyclic aromatic hydrocarbon fraction --- p.84 / Chapter 3.2.1 --- Correlation between heavy metal level and toxicity tests --- p.84 / Chapter 3.2.2 --- Correlation between total polycyclic aromatic hydrocarbon concentration in polycyclic aromatic hydrocarbon fraction and toxicity tests --- p.84 / Chapter 3.2.3 --- Correlation between four acute toxicity tests --- p.88 / DISCUSSION --- p.91 / Chapter 1 --- Chemical analyses of oily wastes --- p.91 / Chapter 1.1 --- Inductively coupled plasma-emission spectrometric analysis --- p.91 / Chapter 1.2 --- Gas chromatography-mass spectrometry analysis --- p.93 / Chapter 2. --- ecotoxicological studies of oily wastes --- p.95 / Chapter 2.1 --- Growth inhibition test on Chlorella pyrenoidosa CU-2 --- p.95 / Chapter 2.2 --- Survival tests on Elasmopus rapax and Parhyale plumulosa --- p.96 / Chapter 2.3 --- Survival test on Ambassis gymnocephalus and Sparus sarba --- p.97 / Chapter 2.4 --- Microtox® test --- p.98 / Chapter 3 --- Statistical analyses of chemical and ecotoxicological analyses --- p.99 / Chapter 4. --- statistical analyses between acute toxicity tests --- p.101 / CONCLUSION --- p.102 / REFERENCES --- p.104

Graduation date: 1995

This dissertation titled ‘An appraisal of the law relating to oil pollution in the inland, territorial and maritime waters of Nigeria’ examines whether the law governing oil pollution in Nigeria is satisfactory in so far as determining issues of liability and compensation for oil pollution are concerned. The thesis examines a research hypothesis on the determination of the question whether the law adequately caters for victims of oil pollution occurring in the inland, territorial and maritime waters of Nigeria and if not, what are the observable defects and how can these defects be remedied. Not only has there been a considerable environmental degradation in Nigeria occasisoned by oil exploration and exploitation, particularly in the areas around the Niger Delta, but there has been serious socio-economic consequences pertinent to sustainable development of Nigeria as a nation. These impacts and the government’s attempts to tackle the problems have been the focus of this thesis. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2012.

Chronic oil pollution or frequent small-scale oil discharges from vessel operations is an important source of marine oil pollution and considered a constant threat to marine and coastal environments. In Canada’s Pacific region, evidence of such illegal discharges has been gathered by the National Aerial Surveillance Program (NASP) from 1998 to 2007. We used this information to fit Generalized Linear Models (GLMs) for offshore waters and inshore waters and explore the relationships between oil spill occurrences and four predictor variables: shipping traffic densities for different vessel types, distance to shore, distance to port and density of small harbours. The best-fit models for both regions show that areas closer to shore and with higher density of small harbours have a higher probability of oil spill occurrences. However, only in inshore waters was shipping traffic significantly related with oil spill occurrences. Tug boats and oil tanker traffic show a significant positive relationship with oil spill observations, while carriers presented a negative association. Mapped results for offshore areas depicted the highest probability of oily discharges in Barkley Sound and at the entrance of the Juan de Fuca Strait; whereas in inshore waters, oil pollution hot spots were found in the vicinity of major commercial and tourist centres. These probability maps were used to identify Coastal and Marine Protected Areas (CMPAs) and Important Bird Areas (IBAs) more likely to be exposed to chronic oil pollution during a period of 10 years. Three areas were highlighted as the most vulnerable based on their likelihood of exposure and the sensitivity of the species they contain to oil pollution. These sites are the Tofino Mudflats, Barkley Sound, Scott Islands and the Sturgeon and Robert Banks, in the Fraser River delta. Our findings provide better understanding of the relationships between oil spill occurrences and vessel operations and help us identify likely oil pollution hot spots and sites particularly vulnerable to this stressor in Canada’s Pacific region. This information can be useful to NASP in improving its efficiency and in targeting monitoring efforts to troublesome areas. Additionally, this research contributes to regional studies that focus on analyzing the distribution of anthropogenic stressors from sea-based activities in British Columbia. Finally, we highlight the importance of collecting accurate data to properly model the probability of oil spill occurrences and encourage future research aiming to better understand and ultimately reduce the chronic release of pollutants from shipping activities into the marine environment.

We explored the combination of metagenomic library and DNA microarray technologies into a single platform as a novel way to rapidly screen metagenomic libraries for genetic targets. In the "metagenomic microarray" system, metagenomic library clone DNA is printed on a microarray surface, and clones of interest are detected by hybridization to single-gene probes. This study represents the initial steps in the development of this technology. We constructed two 5,000-clone large-insert metagenomic libraries from two diesel-contaminated Arctic soil samples. We developed and optimized an automated fosmid purification protocol to rapidly-extract clone DNA in a high-throughput 96-well format. We then created a series of small prototype arrays to optimize various parameters of microarray printing and hybridization, to identify and resolve technical challenges, and to provide proof-of-principle of this novel application. Our results suggest that this method shows promise, but more experimentation must be done to establish the feasibility of this approach.