Thesis (M.Sc. (Microbiology)) --University of Limpopo, 2011 / The leaves of fifteen plant species were collected from the Lowveld Botanical Garden in Nelspruit, Mpumalanga Province, South Africa. The collection was based on a list of plants and their ethnopharmacological information provided by the Phytomedicine Programme at the University of Pretoria. The dried leaves of the plants were powdered and extracted using hexane, dichloromethane, acetone and methanol. The extracts were screened for antibacterial activity against Mycobacterium smegmatis and Rhodococcus erythropolis. The acetone extract of Milletia stulhimannii was the most active, showing activity against Mycobacterium smegmatis and Rhodococcus erythropolis with MIC values 0.13 and 0.08 mg/ml, respectively. Acetone extracts for all plants had the lowest MIC values ranging between 0.11-1.25 mg/ml and 0.08-1.25 mg/ml for M. smegmatis and R. erythropolis, respectively. Milletia stulhimannii, Albizia gummifera, Xanthocercis zambesiaca and Barringtonia racemosa extracts have shown the greatest potential for anti-tubercolosis agents. These were all active against M. smegmatis with an average MIC value of acetone extracts of 0.13 mg/ml. Apodytes dimidiata was selected for the isolation of active compounds since its activity on qualitative antibacterial activity assays was highly prominent on TLC plates in comparison to the other plant extracts. Two compounds were isolated from A. dimidiata but after purification, their MICs were above 2.5 mg/ml indicating a possible loss of activity during purification. The preliminary NMR spectra analysis suggested that the compounds were a long fatty acid and a triterpene. Future work is required to elucidate the chemical structures of the latter compounds and to test the activity of these compounds against Mycobacterium tuberculosis. / Department of Water Affairs, and University of Limpopo Research Development and Administration Office

Thesis (M.Sc. ( Physiology )) --University of Limpopo, 2011 / Refer to document / the National Research Foundation

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The capacity and efficiency of melter slag (provided by New Zealand Steel) to remove heavy metals and suspended solids from stormwater samples are studied in this thesis. A series of batch tests were carried out to investigate the adsorption-desorption mechanism of the slag to remove heavy metals (Cd2+, Zn2+ and Cu2+) from working solutions. The results showed that all the tested metallic ions could be removed by mixing the melter slag with the working solutions. Adsorption and ion exchange are the dominant mechanisms in this process. The adsorption capacity follows the descending order of Cu2+ > Zn2+ = Cd2+. Varied binding energy of different metallic ions to the slag resulted in competitive adsorption between ions.A variety of substances: inorganic salts (KCl, NaCl, KNO3 and sea water), organic acids (citric and tartaric) and inorganic acids (nitric and carbonic), were tested as desorbing agents to recover the used slag. Citric acid in sea water was found to be the best in terms of desorption efficiency and cost-effectiveness.The column tests were carried out to simulate the filtration bed in laboratory conditions. Stormwater samples were collected from residential and commercial areas in Auckland. The synthetic stormwater samples were prepared from clay and metallic ion solution. Both samples were used as feeding solutions in the tests. The removal efficiencies of suspended solids and metallic ions (Zn2+, Cd2+ and Cu2+) vary depending on different operation conditions. They are in inverse proportion to the grain size of the melter slag and the influent flow rate. The higher removal efficiency of Zn2+ than that of suspended solids infers that metallic ions are removed by both precipitation and adsorption.

Estuarine acidification, caused by disturbance of acid sulfate soils (ASS), is a recurrent problem in eastern Australia. Affected waters are characterised by low pH and elevated concentrations of metals, principally aluminium and iron. The effects of acid and elevated metal concentrations associated with ASS, on adult Sydney rock oysters, have not been previously investigated. This study tested links between ASS-affected drainage, subsequent estuarine acidification and Sydney rock oyster production problems on the Hastings and Manning Rivers, mid north coast New South Wales. The primary objective of this thesis was to establish if estuarine acidification causes mortality and slow growth in individual Sydney rock oysters by exposing oysters to low pH, iron and aluminium using field and laboratory experiments. Water quality data showed that estuarine acidification was spatially extensive in the Hastings and Manning Rivers following heavy rainfall and was due to mineral acids originating from drained or excavated ASS. Estuarine acidification regularly affected areas used for Sydney rock oyster production following heavy rainfall. Field experiments showed that Sydney rock oyster mortality rates were significantly higher at sites exposed to ASS-affected waters compared to locations that were isolated from ASS-affected waters. Oyster mortality increased with the time of exposure and smaller oysters (mean weight = 5 g) experienced significantly higher mortality relative to larger oysters (mean weight = 29 g). This was caused by acid-induced shell degradation resulting in perforation of the smaller oysters??? under-developed shells. Additionally, Sydney rock oyster growth rates were dramatically reduced at sites exposed to ASS-affected waters and the overall mean condition index of oysters at ASS-affected field sites was significantly lower than the overall mean condition index of oysters at non-impacted sites. Findings from laboratory experiments showed that ASS-affected water alters oyster valve movements and significantly reduces oyster feeding rates at pH 5.5. Acidic treatments (pH 5.1) containing 7.64 mg L-1 of aluminium or ASS-affected water caused changes in the mantle and gill soft tissues following short-term exposure. Degenerative effects described in oysters in this study were also due to iron contained in ASS-affected waters. Iron precipitates accumulated on the shell, gills and mantle and were observed in the stomach, intestine, digestive tubules and rectum. This study concluded that Sydney rock oysters are unable to tolerate acidic conditions caused by ASS outflows and cannot be viably cultivated in acid-prone areas of the estuary.

The ever-increasing detection of harmful organic and inorganic compounds in habitable areas throughout the world has led to mounting research into applications and techniques for the treatment of contaminated soils, surface and groundwaters, and chemical and industrial wastewaters. Chemical oxidation technologies, in particular Fenton-based remediation systems, have exhibited considerable potential for the effective treatment and remediation of such contaminated waters and soils. The use of Fenton-based oxidation systems for the treatment of contaminated waters and wastewaters warrants the development of kinetic models capable of accurately simulating system behaviour. In this thesis, the kinetics of Fenton-mediated oxidation systems and kinetic models based on its governing reaction mechanism are investigated in order to highlight those parameters and conditions that effect Fenton chemistry and oxidation performance, and to demonstrate the application of such kinetic models to design and improve treatment systems. Experimental and simulated data describing the oxidation of formic acid by Fenton's reagent at low pH (3 to 4) and under a variety of initial conditions, operating regimes, and solution environments supports a proposed reaction mechanism that nominates the hydroxyl radical (OH) as the active oxidizing intermediate in Fenton-based oxidation systems. Laboratory experiments demonstrate that formic acid oxidation is inhibited in the presence of oxygen, and model simulations of these systems reveals that such behaviour is due to the effect organic radical intermediates and/or by-products have in assisting or hindering the redox cycling of the catalytic iron species. The critical role that iron redox cycling plays in affecting oxidation performance is further highlighted by experimental and simulated studies at alternate pHs and using different target organics, including those that react directly with iron in a redox capacity. Experiments at pH 4 reveal an increase in the redox cycling of iron and improved oxidation performance compared to pH 3 as the higher pH favours the superoxide radical, a stronger reductant than the hydroperoxyl radical that predominates at pH 3. Other laboratory and modelling studies on the Fenton-mediated oxidation of certain aromatic compounds highlight the manner in which quinone and quinone-like compounds, being added directly or generated as oxidation by-products, can improve oxidation performance via redox reactions with iron. Further simulations reveal the type of practical design and operating information kinetic models can provide for treatment processes, though it is noted an appropriate understanding of the oxidation mechanism of the target species is necessary for the accurate application of the model.

Cadmium in the ocean has a nutrient-like cycling pattern: with biological uptake at the surface, subsequent sinking in particulate form and then regeneration as dissolved species in deeper waters. Many measurements have been made over time of the ratio of the concentrations of dissolved Cd to those of PO₄ (Cd/PO₄) in the world ocean and this has become one of the best relationships documented between a trace metal and a nutrient. Combined with the measurements of the Cd/Ca ratio in foraminifera, the Cd/PO₄ ratio has been used to reconstruct the oceanographic circulation patterns that existed during past glacial periods and hence provides information on past climate changes. In the present study Cd/PO₄ ratios of the Southern Indian Ocean in surface and deep waters were investigated. The slopes of the relationships between Cd and PO₄ concentrations in waters of this region are high compared to the global correlations, and lie between those reported for other parts of the Southern Ocean and the Pacific Ocean. In surface waters of the Southern Indian Ocean, Cd/PO₄ ratios decrease from regions exhibiting high nutrient-low chlorophyll (HNLC) characteristic in the south to oligotrophic waters further north. It is also found that particulate Cd plays an important role in regulating the high Cd/PO₄ ratios reported in waters south of the Polar Front. Very low Cd/PO₄ ratios were measured in waters associated with the Subtropical Front southeast of New Zealand compared to other Southern Ocean and global oceanic waters. Seasonal variations in the Cd/PO₄ ratios measured for these waters strongly suggest they are associated with a significant biological uptake of dissolved Cd particularly during the phytoplankton growth season in summer. Dissolved Fe concentrations in the Southern Indian Ocean and seasonal variations of Fe in waters off the Otago Coast (southeast of New Zealand) suggest that Fe may stimulate phytoplankton growth and this might result in lower Cd/PO₄ ratios in surface waters through enhanced Cd uptake relative to PO₄ by the phytoplankton. However there is no distinct relationship between dissolved Fe concentrations and the dissolved Cd/PO₄ ratios measured in these surface waters. This finding is in disagreement with the recent 2006 hypothesis put forward by J.T. Cullen, which proposed that waters exhibiting low dissolved Cd/PO₄ ratios were associated with the HNLC regions. From a consideration of the potential Zn concentrations calculated from Si concentration measurements reported for these waters, it would appear that Zn may play a more important role than Fe in regulating Cd/PO₄ ratios in these waters. Measurements of dissolved and total Cd concentrations relative to those of PO₄ were also undertaken in the Otago Harbour and immediate surrounding coastal waters. These exhibited higher Cd concentrations and higher Cd/PO₄ ratios than open ocean waters further off the Otago Coast. The particulate Cd concentrations showed a negative correlation with Cd concentrations measured in cockle species (Austrovenus stuchburyi) collected in the harbour, suggesting that particulate Cd is not the source of Cd measured in the tissue of this species. The concentrations of Cd and other trace metals were also measured in samples of green mussel (Perna canaliculus), ribbed mussel (Aulacomya atra maoriana) and oyster (Saccostrea cucullata) collected from Otago Harbour and possible correlations explored between these concentrations and other parameters such as the shellfish condition indices and environmental gradients in the harbour. In summary, measurements of dissolved and particulate Cd concentrations in the water column can provide unique information on a number of processes occurring in the global marine environment.

Levels of ⁶⁵Zn in water, sediments, plants, and animals in Alder Slough, a small ecosystem in the Columbia River Estuary, were determined periodically during a one year period. Concentrations of total Zn in organisms were also measured to permit computation of specific activities (μCi⁶⁵Zn/g total Zn). Temporal fluctuations in the activities of ⁶⁵Zn and the concentrations of total Zn occurred in all components of the ecosystem. Activities of ⁶⁵Zn and concentrations of total Zn varied among individual fish from the study area. Pooled estimates of population standard deviations of these values in all samples of two fish species taken during the year amounted to about 20% of their grand means. Usually, individuals with high ⁶⁵Zn activities also proved to have high total Zn concentrations, hence specific activities were less variable than either ⁶⁵Zn or total Zn. The nuclear reactors at Hanford, Washington, the source of most of the ⁶⁵Zn in the Columbia River, were shut down for approximately 45 days during this study. This event presented the opportunity to investigate the response of various components of the ecosystem to reduced ⁶⁵Zn input. The rates of ⁶⁵Zn specific activity decline in several organisms were measured following reactor shutdown. The time required for specific activity of an organism to be reduced by one-half under the conditions prevailing is termed "ecological half-life". The "ecological half-life" differs from the biological half-life of an organism because: 1. it is defined in terms of specific activity 2. it reflects the continued addition of radioactivity to the organism from its food web and water. "Ecological half-lives" vary with the trophic levels of the organisms and with changes in prevailing ecological conditions. / Graduation date: 1968

Contamination of recreational and shellfish waters with fecal pollution is a major water quality issue with associated economic impacts and human health risks. Reliable fecal source identification and rapid, quantitative analyses are essential components of risk assessment. Enteric bacteria that are endemic to specific hosts have a potential role as public health indicators of fecal pollution. Building on previous work to discriminate ruminant and human fecal contamination, we cloned class Bacteroidetes 16S rRNA genes from pig, elk, dog, cat, and seagull fecal DNAs. Unique restriction patterns were identified among clones from each of the host species using Terminal Restriction Fragment Length Polymorphisms (T-RFLP). Clones exhibiting unique patterns were sequenced and analyzed phylogenetically, along with human, horse, and cattle sequences recovered from previous work. The analysis revealed both endemic and cosmopolitan (global) host distributions. The sequence data were used to identify host-specific genetic markers for pig and horse feces, and to design PCR primers that identify these sources of fecal pollution in water. There was a high degree of sequence overlap among the fecal Bacteroidetes of wild and domestic ruminants, and among human, domestic pet, and seagull Bacteroidetes. We compared fecal Bacteroidetes rRNA genes from these hosts using subtractive hybridization, a method that identifies differences between closely related genomes or gene sequences. A Bacteroidetes rDNA marker that distinguishes elk and cow feces was identified, as well as a host-specific marker for dog fecal Bacteroidetes. The four newly designed PCR primers were tested for specificity and sensitivity, and the dog primer was successfully used, along with the human and ruminant-specific primers, in a collaborative study comparing fecal source tracking methods. We also developed a real time Taq nuclease assay for quantification of fecal Bacteroidetes 16S rDNA, and compared it with an EPA-approved enumeration method for the current standard public health indicator, Escherichia coli, in serial dilutions of sewage primary influent. There was a strong, positive correlation between the methods, and the Taq nuclease assay was sensitive and much more rapid than the E. coli assay. PCR source identification and enumeration of fecal Bacteroidetes 16S rDNA show promise for application in a health risk-based analysis of fecal pollution. / Graduation date: 2005