by Ma Suet-yee. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 119-131). / Abstracts in English and Chinese. / Acknowledgment --- p.i / Abstract --- p.ii / 摘要 --- p.iv / Table of Contents --- p.vi / List of Tables --- p.x / List of Figures --- p.xii / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Ganoderma lucidum --- p.1 / Chapter 1.1.1 --- History of Ganoderma lucidum --- p.1 / Chapter 1.1.2 --- Classification --- p.1 / Chapter 1.1.3 --- Macroscopic and microscopic structure --- p.2 / Chapter 1.1.4 --- Ganoderma lucidum as a pathogen --- p.3 / Chapter 1.1.5 --- Availability of tree hosts in Hong Kong --- p.4 / Chapter 1.1.6 --- Medicinal effects --- p.5 / Chapter 1.2 --- Study of Populations in Fungi --- p.6 / Chapter 1.2.1 --- Definition of Population --- p.6 / Chapter 1.2.2 --- Study of Fungal Populations --- p.7 / Chapter 1.2.3 --- Techniques for Population Studies in Fungi --- p.7 / Chapter 1.2.3.1 --- Somatic Incompatibility Test / Chapter 1.2.3.2 --- Isozyme Analysis / Chapter 1.2.3.3 --- Restriction Fragment Length Polymorphisms (RFLPs) / Chapter 1.2.3.4 --- Polymerase Chain Reaction (PCR) Amplification / Chapter 1.3 --- Mitochondrial DNA (mt-DNA) in Fungi --- p.14 / Chapter 1.3.1 --- Inheritance in mt-DNA --- p.15 / Chapter 1.3.2 --- Mitochondrial DNA in Population Studies --- p.15 / Chapter 1.3.2.1 --- Mitochondrial small-subunit (mt-SSU) rDNA / Chapter 1.3.2.2 --- Cytochrome oxidase 3 (cox3) / Chapter 1.4 --- Biodiversity study on Ganoderma species --- p.19 / Chapter 1.5 --- Environment Pollutants in Hong Kong --- p.20 / Chapter 1.5.1 --- Air quality in Hong Kong --- p.20 / Chapter 1.5.2 --- Soil quality in Hong Kong --- p.20 / Chapter 1.5.3 --- Toxicity of pollutants --- p.23 / Chapter 1.5.4 --- Accumulation of heavy metals by G. lucidum --- p.26 / Chapter 1.6 --- Objectives of Study --- p.27 / Chapter 1.7 --- Project Strategies --- p.28 / Chapter 1.7.1 --- Survey on distribution and collection of Ganoderma lucidum in Hong Kong --- p.28 / Chapter 1.7.2 --- Genetic divergences of G. lucidum mitochondrial genes --- p.28 / Chapter 1.7.3 --- Contaminations on field collected G. lucidum --- p.29 / Chapter 1.8 --- Significance of Study --- p.29 / Chapter Chapter 2 --- Materials and Methods --- p.30 / Chapter 2.1 --- Collection of Ganoderma lucidum in Hong Kong --- p.30 / Chapter 2.2 --- Tissue Isolation --- p.30 / Chapter 2.3 --- Somatic Incompatibility Test --- p.36 / Chapter 2.4 --- Molecular Identification --- p.40 / Chapter 2.4.1 --- Extraction of DNA --- p.40 / Chapter 2.4.2 --- Gel electrophoresis --- p.41 / Chapter 2.4.3 --- Strain authentication by arbitrarily primed polymerase chain reaction (APPCR) --- p.41 / Chapter 2.4.4 --- PCR of mt-SSU rDNA and --- p.43 / Chapter 2.4.5 --- Sequencing of mt-SSU rDNA and cox3 --- p.44 / Chapter 2.4.6 --- Comparison of G. lucidum complex with other Ganoderma and related species / Chapter 2. 4.7 --- Phylogenetic analyses --- p.46 / Chapter 2.5 --- Investigation of pollutants in Ganoderma lucidum collected in Hong Kong --- p.46 / Chapter 2.5.1 --- Metal analysis --- p.48 / Chapter 2.5.1.1 --- Acid digestion / Chapter 2.5.1.2 --- Statistical analysis / Chapter 2.5.2 --- Organic pollutant analysis --- p.49 / Chapter Chapter 3 --- Result --- p.52 / Chapter 3.1 --- Collection of Ganoderma lucidum in Hong Kong --- p.52 / Chapter 3.1.1 --- Field observation --- p.52 / Chapter 3.1.2 --- Macroscopic characteristics --- p.52 / Chapter 3.1.3 --- Microscopic characteristics --- p.53 / Chapter 3.2 --- Somatic Incompatibility Test --- p.56 / Chapter 3.3 --- DNA fingerprints by Arbitrarily-Primed PCR --- p.57 / Chapter 3.4 --- Sequencing of mt-SSU rDNA region of G. lucidum and related species --- p.60 / Chapter 3.4.1 --- Genetic variability in mt-SSU rDNA region of G. lucidum --- p.60 / Chapter 3.4.2 --- mt-SSU rDNA region of G. lucidum and other related species --- p.61 / Chapter 3.4.3 --- Phylogenetic analysis of mt-SSU rDNA region --- p.61 / Chapter 3.5 --- Sequencing of cox3 region --- p.71 / Chapter 3.5.1 --- Genetic variability in cox3 region of G. lucidum --- p.71 / Chapter 3.5.2 --- cox3 region of G. lucidum and other related species --- p.72 / Chapter 3.5.3 --- Phylogenetic analysis of cox3 region --- p.72 / Chapter 3.6 --- Metal content of field G. lucidum --- p.82 / Chapter 3.7 --- Organic pollutants in field collected G. lucidum --- p.90 / Chapter Chapter 4 --- Discussion --- p.93 / Chapter 4.1 --- Collection of Ganoderma lucidum in Hong Kong --- p.93 / Chapter 4.1.1 --- Differentiation of G. lucidum and related species in the G lucidum species complex --- p.93 / Chapter 4.1.2 --- Field observation --- p.94 / Chapter 4.2 --- Biodiversity of populations of G. lucidum in Hong Kong --- p.95 / Chapter 4.2.1 --- Individualism of G. lucidum --- p.95 / Chapter 4.2.2 --- Genetic variability in mt-SSU rDNA region of G. lucidum --- p.96 / Chapter 4.2.3 --- Genetic variability in cox3 region of G. lucidum --- p.98 / Chapter 4.2.4 --- Lineages of G. lucidum collected in Hong Kong --- p.100 / Chapter 4.2.5 --- Cryptic phylogenetic species --- p.101 / Chapter 4.3 --- Contamination of field collected Ganoderma lucidum in Hong Kong --- p.106 / Chapter 4.3.1 --- Metal contents in field collected G. lucidum in Hong Kong --- p.106 / Chapter 4.3.1.1 --- Metal contents of G. lucidum fruiting bodies collected at each site / Chapter 4.3.1.2 --- General discussion of metals / Chapter 4.3.1.3 --- Consumption of field collected G. lucidum fruiting bodies / Chapter 4.3.2 --- Comparison of metal contents between field collected Hong Kong G. lucidum with other mushrooms collected from other places --- p.112 / Chapter 4.3.3 --- Survey of organic pollutants in field collected G. lucidum in Hong Kong --- p.113 / Chapter Chapter 5 --- Conclusion --- p.116 / Chapter Chapter 6 --- Further investigation --- p.118 / Chapter Chapter 7 --- Reference --- p.119

by Wong Pik-kwan. / Thesis submitted in: September 1997. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references (leaves 116-139). / Abstract also in Chinese. / ABSTRACT --- p.i / ACKNOWLEDGEMENT --- p.iii / TABLE OF CONTENTS --- p.iv / LIST OF TABLES --- p.viii / LIST OF FIGURES --- p.ix / Chapter CHAPTER ONE --- GENERAL INTRODUCTION --- p.1 / Chapter CHAPTER TWO --- LITERATURE REVIEW / Chapter 2.1 --- Introduction --- p.4 / Chapter 2.2 --- Heavy metals --- p.6 / Chapter 2.3 --- Mechanisms of metal toxicity --- p.9 / Chapter 2.4 --- Toxic effects of metals on marine organisms --- p.10 / Chapter 2.4.1 --- Cadmium --- p.10 / Chapter 2.4.2 --- Chromium --- p.10 / Chapter 2.4.3 --- Copper --- p.11 / Chapter 2.4.4 --- Lead --- p.12 / Chapter 2.4.5 --- Nickel --- p.12 / Chapter 2.4.6 --- Zinc --- p.13 / Chapter 2.5 --- Metal uptake and elimination in marine organisms --- p.14 / Chapter 2.5.1 --- Uptake of metals --- p.14 / Chapter 2.5.2 --- Elimination of metals --- p.15 / Chapter 2.5.3 --- Metal detoxification system in fish --- p.16 / Chapter 2.6 --- Heavy metals in marine fish --- p.17 / Chapter 2.7 --- Bioaccumulation --- p.20 / Chapter 2.7.1 --- Models of metal accumulation --- p.21 / Chapter 2.7.2 --- Compartment model --- p.21 / Chapter 2.7.3 --- Physiologically based pharmacokinetic (PB-PK) model --- p.22 / Chapter 2.8 --- The influence of environmental factors on bioaccumulation of metals --- p.23 / Chapter 2.8.1 --- Temperature --- p.23 / Chapter 2.8.2 --- Salinity --- p.23 / Chapter 2.8.3 --- Organic matter --- p.24 / Chapter 2.8.4 --- pH --- p.25 / Chapter 2.8.5 --- Chelators and surfactants --- p.25 / Chapter 2.8.6 --- Other metals --- p.26 / Chapter 2.9 --- Biological effects of heavy metals on man --- p.26 / Chapter 2.10 --- The use of biological indicator organisms for metal pollution --- p.28 / Chapter CHAPTER THREE --- HEAVY METAL CONCENTRATIONS IN CULTURED MARINE FISH IN HONG KONG / Chapter 3.1 --- Introduction --- p.31 / Chapter 3.2 --- Materials and methods --- p.36 / Chapter 3.2.1 --- Sampling --- p.36 / Chapter 3.2.2 --- Water analysis --- p.36 / Chapter 3.2.3 --- Sediment analysis --- p.39 / Chapter 3.2.4 --- Mussel analysis --- p.40 / Chapter 3.2.5 --- Fish analysis --- p.40 / Chapter 3.2.6 --- Quality control and statistical analysis --- p.41 / Chapter 3.3 --- Results --- p.42 / Chapter 3.3.1 --- Seawater --- p.42 / Chapter 3.3.2 --- Sediment --- p.46 / Chapter 3.3.3 --- Mussel --- p.46 / Chapter 3.3.4 --- Fish --- p.50 / Chapter 3.4 --- Conclusion --- p.67 / Chapter 3.4.1 --- "Metal concentration in seawater, sediment, green mussel and fish" --- p.67 / Chapter 3.4.2 --- Accumulation of heavy metals in different tissues of cultured fish --- p.69 / Chapter 3.4.3 --- Relationship between the body weight and metal accumulation --- p.71 / Chapter 3.4.4 --- Heavy metal pollution in fish culture sites --- p.72 / Chapter 3.4.5 --- Selection of fish culture site --- p.72 / Chapter CHAPTER FOUR --- ACUTE AND SHORT-TERM EFFECTS OF COPPER(II) IONS ON SPARUS SARBA / Chapter 4.1 --- Introduction --- p.76 / Chapter 4.2 --- Materials and methods --- p.79 / Chapter 4.2.1 --- Experimental animals --- p.79 / Chapter 4.2.2 --- Determination of the 96 hour median lethal concentrations --- p.19 / Chapter 4.2.3 --- Determination of growth rate --- p.80 / Chapter 4.3 --- Results --- p.82 / Chapter 4.3.1 --- Determination of the 96 hour median lethal concentrations --- p.82 / Chapter 4.3.2 --- Determination of growth rate --- p.82 / Chapter 4.3.3 --- Distribution of Cu concentration in whole body and different tissues of S. sarba --- p.82 / Chapter 4.4 --- Conclusion --- p.91 / Chapter 4.4.1 --- Determination of the 96 hour median lethal concentrations --- p.91 / Chapter 4.4.2 --- Determination of growth rate --- p.93 / Chapter 4.4.3 --- Distribution of Cu concentration in whole body and different tissues of S. sarba --- p.94 / Chapter CHAPTER FIVE --- ACCUMULATION AND ELIMINATION OF COPPER(II) IONS TO SPARUS SARBA / Chapter 5.1 --- Introduction --- p.96 / Chapter 5.2 --- Materials and methods --- p.98 / Chapter 5.2.1 --- Experimental animals --- p.98 / Chapter 5.2.2 --- Uptake and elimination of Cu ion in S. sarba during continuous exposure to waterborne Cu --- p.98 / Chapter 5.3 --- Results --- p.100 / Chapter 5.4 --- Conclusion --- p.108 / Chapter CHAPTER SIX --- GENERAL CONCLUSION --- p.112 / CHAPTER SEVEN REFERENCES --- p.116

Graduation date: 1993

M.Sc. / The Vaal Dam subcatchment is located in the upper reaches of the Vaal River. As a result the water quality is reasonably good. In contrast the Vaal River Barrage catchment includes the PWV area, resulting in poorer water quality. During this study, a fish Health Assessment Index (HAI) successfully tested in previous studies on the Olifants River System was applied to the Vaal River System. The aim was to determine if the HAI could distinguish between the Vaal Dam and Vaal River Barrage with regards to water quality. Surveys were conducted bimonthly from November 1998 to February 2000. Physical water quality variables were measured. Water and sediment samples were also collected and analyzed (Institute for Water Quality Studies, Department of Water Affairs and Forestry) to verify the HAI results. Rand Water Board and the Department of Water Affairs and Forestry also made water quality data available. Twenty sharptooth catfish, Clarias gariepinus, were collected from both localities with the aid of gill nets. Fish were checked for external parasites on the boat. On land fish were weighed and measured, after which blood was drawn and slime smears made. Blood and slime smears were examined under a light microscope for parasites. The HAI examination was performed after severing the spinal cord. Internal parasite numbers were recorded. From the parasite data collected infestation statistics were calculated. Four variations of the Parasite Index (PI) were incorporated in the HAI and results compared. During each survey gill arch, gill filament, muscle, skin and liver tissues were collected from each fish. These were analyzed for strontium, aluminium, chromium, manganese, iron, lead, copper, zinc and nickel concentrations using atomic absorption spectrophotometry. Differences in water and sediment trace metal concentrations between localities were small. Metal concentrations in fish tissues recorded from both localities were also almost identical. Possible explanations for this trend were discussed in the relevant section. Highest metal concentrations were generally recorded in gill tissue followed by liver, skin and muscle. Physical water quality variables (salinity and conductivity), and macro water analysis (e.g. phosphate and nitrate) indicated that water quality at the Vaal River Barrage was poorer. The HAI confirmed this. Higher index values were recorded from the Vaal River Barrage, with the converse being true for the Vaal Dam. Regression analysis indicated that plasma protein, haematocrit and the index value obtained using the Inverted Parasite Index, most successfully predicted (70 %) from where a randomly chosen fish were collected. When viewing index values obtained using the four versions of the PI, all four distinguished between localities. The discriminatory ability of the Inverted PI was slightly higher than that of the other PI's. It is concluded that the HAI distinguished successfully between the Vaal Dam and Vaal River Barrage on the grounds of water quality. Poor fish health correlated with decreasing water quality (salinity and eutrophication).

Biosorption is a property of certain type of inactive, microbial biomass to bind and concentrate heavy metals from even very dilute aqueous solutions. Biomass exhibits this property, acting just as a chemical substance, as an ion exchanger of biological origin. It is particularly the cell wall structure of certain algae that is found responsible for this phenomenon. In these experiments, the rate and extent for removal of copper is subjected to parameters such as pH, initial metal concentration, biosorbent size, contact time, temperature and the ability of the biomass to be regenerated in sorption-desorption experiments. The metal adsorption was found to be rapid within 25 minutes. The maximum copper uptake of 30 mg of copper / g of biomass has been observed, in the following conditions: 100 mg / L, 0.1 g of biomass, pH 4 and at temperature of 25°C. From this study, it was found that copper uptake is increasing with increase in pH, with optimum being pH 4. Copper uptake increases substantially from 0 to 25 minutes. Metal biosorption behaviour of raw seaweed Sargassum in six consecutive sorptiondesorption cycles were also investigated in a packed-bed column, during a continuous removal of copper from a 35 mg/l aqueous solution at pH 4. The sorption and desorption was carried out for an average of 85 and 15 hours, respectively, representing more than 40 days of continuous use of the biosorbent. The weight loss ofbiomass after this time was 13.5%. The column service time decreased from 25 hrs in the first cycle to 10 hrs for the last cycle. / Thesis (M.Sc.)-University of KwaZulu-Natal, 2005.

Submitted in partial fulfillment of the requirements for the degree in Masters of Technology: Chemistry, ML Sultan Technikon, Durban, 2003. / The pollution of water bodies by heavy metals is a serious threat to humanity. The technique known as phytoremediation is used to clean up these polluted water bodies. The accumulation of heavy metals by aquatic plants is a safer, . cheaper and friendlier manner of cleaning the environment. The aquatic plants -studied in this project are A.sessilis, P.stratiotes, R.steudelii and T.capensis. The accumulation of heavy metals in aquatic plants growing in waste water treatment ponds was investigated. The water, sludge and plants were collected from five maturation ponds at the Northern Waste Water Treatment Works, Sea Cow Lake, Durban. The samples were analysed for Zn, Mn, Cr, Ni, Pb and Cu using ICP-MS. In general it was found that the concentrations of the targeted metals were much lower in the water (0.002 to 0.109 mg/I) compared to sediment/sludge (44 to 1543mg/kg dry wt) and plants (0.4 to 2246 mg/kg dry wt). These results show that water released into the river from the final maturation pond has metal concentrations well below the maximum limits set by international environmental control bodies. It also shows that sediments act as good sinks for metals and that plants do uptake metals to a significant extent. Of the four plants investigated it was found that }t.sessi[ir (leaves, roots and stems) and }A.sessilis (roots and stems) are relatively good collectors of Mn and Cu respectively. These findings are described in the thesis. The concentration of heavy metals in the stems, leaves and roots of the three plants were compared to ascertain if there were differences in the ability of the plant at different parts of the plant to bioaccumulate the six heavy metals studied. / M

M.Sc. / South Africa is renowned for its exploitable mineral resources and continues to be a major player in the world’s mineral markets. The country is well known for containing the world’s largest gold and platinum repositories and electroplating industries, which is the major cause for delivering by-products such as cadmium (Cd), chromium (Cr) and nickel (Ni). Environmental pollution caused by active mining and seepage from closed mines, continuously threatens South African water resources. Such pollution can cause a shift in water chemistry and increase the availability of certain metals to the living organisms of such a system. Even at low concentrations metals are amongst the most toxic environmental pollutants. As a result of their persistence and capacity to accumulate in the environment, metals have a lasting detrimental effect on the ecosystem. Although there is progress in the treatment of metallic wastes, the discharge thereof by industries is still a serious water pollution problem. In the past, chemical analysis of water has proven to be of great use for the detection of pollutants within the environment. The value of chemical analysis alone has become limiting, as chemical analysis supplies information on the levels of chemicals at a certain time. Furthermore, the monitoring of water quality variables often does not reflect long-term events that may play a critical role in determining the ecosystem health. It is now generally understood that measurements of only the physical and chemical attributes of water cannot be used as surrogates for assessing the health of an aquatic ecosystem. The new trend is to incorporate biological monitoring into Abstract existing monitoring strategies. Fish are entirely dependent on the aquatic environment for their survival, rendering them a good monitor of water pollution. Macroscopic changes in organs are preceded by changes at the tissue, cellular or molecular level. These changes are the net result of adverse biochemical and physiological changes within an organism. Histological analysis is a therefore very sensitive parameter and a valuable technique in determining cellular changes in target organs as a result of exposure to stressors. Fish histology can thus be used as an indicator of exposure to contaminants and assess the degree of pollution. Because of the subjective nature of morphological studies correlations with other quantitative studies are difficult. However, incorporation of quantitative methods is essential to the continued development of histopathology as a biomarker of pollution exposure, and to the interpretation of histological responses. The aim of this study is to qualitatively and quantitatively describe the toxic induced histological changes in the selected organs of Oreochromis mossambicus after acute exposure to Cd, Cr and Ni. Fish were exposed to 10% (n=20) and 20% (n=20) of the LC50 concentration of Cd, Cr and Ni respectively under controlled conditions (23 ± 1°C) for 96 hours in an environmental room with a control group (n=5) for each exposure.

M.Sc. / This study aims to assess the potential impacts of intense feedlot activity on the aquatic freshwater environment, with reference to three feedlots, ranging in production size and all situated in the upper Vaal catchment area. Field assessments were done over a high flow and low flow period, while controlled exposures were also done to quantify a potential stress reaction to growth hormone exposure (using Clarias gariepinus as test organism). It was ascertained that water quality variables contributing towards differences between upstream and downstream environmental conditions are NH4 concentrations pH and conductivity. Lead concentrations were also periodically higher downstream from feedlot activity, in comparison with upstream. Taking the sediment assimilation potential of growth hormones into consideration, it was determined that Feedlot C showed the highest assimilation potential, while Feedlot A reflected the lowest. Alterations on family level invertebrate community structures indicated a categorical decline in abundances and species richness at sites situated downstream from feedlots. However, some clear seasonal influences were also observed. Further community and diversity analyses reflected alterations in invertebrate community structures that were not reflected in SASS 5 scores. With regards to the biomarkers applied in this study, it was noted that there was a significant (p<0.05) difference in the cellular energy allocation (CEA) between control and hormone exposed groups. The total amount of energy available (Ea) increased significantly for test organisms exposed to Diethylstilbestrol (DES), while there was a significant increase in energy consumption (Ec) of test organisms exposed to Trenbolone acetate (TBA). In addition to CEA, metabolic profiling of blood plasma was also performed, which indicated a definite ordination in metabolic constituents after fifteen days of exposure. This was established by subjecting the data to principle component analysis (PCA), which accounted for 83 % variance observed. The impacts and biotic responses identified in this study were contextualised with known literature on the effects of feedlot activity and growth hormone exposure on the aquatic environment. Finally, conclusions were drawn and recommendations made with regard to improving feedlot operational activities. The results obtained in this study contribute towards an integrated framework for the environmental management of feedlot activities.

M.Sc. (Zoology) / Please refer to full text to view abstract

Thesis (MTech (Environmental Health))--Cape Peninsula University of Technology, 2017. / Surface water is used as a source of water supply in many countries, including South Africa. One of the sources of surface water pollution is leachate and surface runoff from landfills. In agricultural soils, the landfill runoff and leachate deteriorate the quality and affect the fertility of soil. The entry of metals and microorganisms from landfill leachate to adjacent environments is through surface runoff due to rainfall. Adverse effects on human- and environmental health triggers a need to monitor and control contaminants in the environment. The aims of the study are to determine the effect of landfill runoff and leachate on agricultural soil and river water (Veldwachters River) running adjacent to the Devon Valley landfill site and to identify potential metal-tolerant organisms in environmental samples collected in Stellenbosch, Western Cape, South Africa. Samples (agricultural soil, river water and sediments) were collected once a month for a period of six months from the study area for analysis. Physicochemical parameters that are known to have major effects on environmental samples were assessed and the concentrations of various metals (Al, Pb, Cr, Mn, Mo, Co, Ni, Cu, Zn, Fe, Cd and V) were also determined by means of inductively coupled plasma mass spectrometry (ICP-MS). Soil texture analysis was tested in order to monitor the metal distribution in soils under the influence of environmental factors.