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Thesis (M. Tech.: Biotechnology)-Dept. of Biotechnology and Food Technology, Durban University of Technology, 2008. xiv, 103 leaves : ill. / Phytoremediation is an emerging technology where specially selected and engineered metal-accumulating plants are used for bioremediation. Amaranthus dubius (marog or wild spinach) is a popular nutritious leafy vegetable crop which is widespread especially in the continents of Africa, Asia and South America. Their rapid growth and great biomass makes them some of the highest yielding leafy crops which may be beneficial for phytoremediation. This study was undertaken to evaluate the potential of A. dubius for the phytoremediation of Chromium (Cr), Mercury (Hg), Arsenic (As), Lead (Pb), Copper (Cu) and Nickel (Ni). Locally gathered soil and plants of A. dubius were investigated for the metals from a regularly cultivated area, a landfill site and a sewage site. Metals were extracted from the samples using microwave-digestion and analyzed using Inductively Coupled Plasma – Mass Spectroscopy (ICP-MS). Further experiments were conducted with plants from locally collected seeds of A. dubius, in a tunnel house under controlled conditions. The mode of phytoremediation, the effect of the metals on the plants, the ability of the plant to extract metals from soil (Bioconcentration Factor - BCF), and the ability of the plants to move the metals to the aerial parts of the plants (Translocation Factor - TF) were evaluated for the different metals. Finally, A. dubius was micro-propagated in a tissue culture system with and without exposure to the metal, and the effect was studied by electron microscopy.

Microorganisms have the potential to remove heavy metals from polluted waters and effluents and may be used in clean-up processes. Microbial associations were enriched for and adapted to grow in nutrient solutions containing various concentrations of different metals. As immobilised cells are known to be more stable and more efficient in metal uptake than are corresponding planktonic or free-living cells the attachment of the microbial associations was investigated using a model stream and it was found that biofilm development was better on rough surfaces such as ground glass and polystyrene than on smooth surfaces such as unetched glass plates and glass beads. When comparing metal uptake by planktonic and attached microorganisms, attached populations were found to have a greater metal-uptake capacity. The uptake of individual metals from various metal combinations was tested with various proportions of pregrown metal-adapted microbial populations as inoculum and it was found that a particular metal was taken up more readily by microbial associations which had previously been exposed to that metal. Lead (Pb2+) appeared to be taken up more readily than copper (Cu2+) or cadmium (Cd2+) while Cd2+ was more actively removed than Cu2+ from solution. pH also affected metal uptake and the optimum range for Cu2+ uptake by the Cu2+ -adapted microbial association was found to be between 5.8 and 7.0. Dead microbial biomass was investigated and found to have efficient metal uptake capacity. Living mycelium from an isolated Aspergillus species showed poor uptake of Cu2+ initially, but when this fungus was pregrown and subsequently killed by moist heat treatment the non-living mycelium was efficient in removal of Pb2+ and Cu2+ ions. The optimum mycelial biomass concentration for metal uptake was also determined. The mechanism of metal uptake by this Aspergillus species was determined, using electron microscopy and EDX techniques, to be metabolism-independent biosorption onto the hyphal surface. Thus the microbial associations and fungal cultures used in this study were shown to have the potential for use in the removal of heavy metals from polluted waters. / Thesis (Ph.D.)-University of Natal, Pietermartizburg, 1995.

With the exceptions of mixing and heating mechanisms, and the recycling of settled solids, no radical changes or improvements have been made to conventional anaerobic digesters treating municipal sewage. These digesters usually function with a hydraulic retention time of 30 to 60 days and at a total solids concentration of 2.6 %(m/v). Volumetric loading is limited since high loadings effect the displacement of the slow growing methanogens. Thus, the hydraulic retention time is coupled to the solids retention time. A crossflow microfiltration unit has been constructed at Northern Waste Water Treatment Works, Durban, to concentrate sludge from a conventional anaerobic digester and, thus, facilitate operation with a higher solids concentration. In addition, this process should result in the retention of the active biomass which would otherwise be lost as a waste product of the treatment process. The solids retention time is, thus, decoupled from the hydraulic retention time. The net result could be higher volumetric loadings, increased microbial activity and increased volatile solids destruction and, hence, improvement in the efficiency of anaerobic digestion of sewage sludge. To test these, different experiments were conducted to specifically determine the effect of higher solids loads. Preliminary experiments were undertaken to determine the biodegradability of primary sludge from the Northern Waste Water Treatment Works. Results showed that primary sludge of 76% VS could be reduced to approximately 48 to 50% VS during an experimental period of 85 days. Reduction of the first 20% VS was rapid if conditions were optimum but subsequent reduction from 55 to 50% VS was slow. It was calculated that approximately 0.88 l gas was produced for every g volatile solids catabolised. Further experiments were conducted to investigate the effects of different solids concentrations on microbial activity. The results showed that the volume of gas produced increased as the solids concentration increased from 2 to 6%(m/v). Digesters with solids concentrations of 6 to 13%(m/v) produced similar volumes of gas. Digesters with solids concentrations of 6 to 13%(m/v) TS produced approximately 300 ml more gas than the control during the 20 days experimental period. The rate of gas production also increased as the solids concentration increased. However, digesters containing 11%(m/v) and 13%(m/v) TS produced similar rates. These results indicate that the introduction of concentrated sludge into the digester improves digestion efficiency. Finally, a semi-continuous digester was operated at a 30 days retention time and at optimum temperature to investigate the efficacy of digesters with increased solids concentrations. The results showed that the rate of gas production increased as the solids concentration increased from 2%(m/v)(control) to 3.8%(m/v). However, the digester operated with 4.7%(m/v) TS produced gas at a rate lower that the digester with 3.8%(m/v) TS. The volatile solids concentrations of all four digesters were similar, indicating neither favourable nor unfavourable effects from increased solids concentrations. The digesters operated with 3.8%(m/v) and 4.7%(m/v) TS produced higher concentrations of volatile acids than the control. The alkalinity concentrations (>_4000 mg t-1 ) were similar for all four digesters. / Thesis (M.Sc.)-University of Natal, Durban, 1995.

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The indiscriminate use of fossil fuels has led to global problems of greenhouse gas emissions, environmental degradation and energy security. Developments of alternative and sustainable energy resources have assumed paramount importance over the past decades to curb these challenges. Biohydrogen is emerging as an alternative renewable source of energy and has received considerable attention in recent years due to its social, economic and environmental benefits. It can be generated by dark fermentation on Organic Fraction of Solid Municipal Waste (OFSMW). These OFSMW exist abundantly and poses disposal challenges. This study models and optimizes the production of biohydrogen on a mixture of agro-municipal wastes; it examines a semi-pilot scale production on these substrates and the feasibility of generating bioelectricity from the process effluents and reviews the prospect of enhancing fermentative biohydrogen development using miniaturized parallel bioreactors. The fermentation process of biohydrogen production on agro-municipal wastes was modelled and optimized using a two-stage design. A mixture design was used for determination of optimum proportions of co-substrates of Bean Husk (BH), Corn Stalk (CS) and OFSMW for biohydrogen production. The effects of operational setpoint parameters of substrate concentration, pH, temperature and Hydraulic Retention Time (HRT) on hydrogen response using the mixed substrates were modelled and optimized using box-behnken design. The optimized mixtures were in the ratio of OFSMW: BH: CS = 30:0:0 and OFSMW: BH: CS = 15:15:0 with yields of 56.47 ml H2/g TVS and 41.16 ml H2/g TVS respectively. Optimization on physico-chemical parameters using the improved substrate suggested optimal setpoints of 40.45 g/l, 7.9, 30.29 oC and 86.28 h for substrate concentration, pH, temperature and HRT respectively and hydrogen yield of 57.73 ml H2/g TVS. The quadratic polynomial models from the mixture and box-behnken design had a coefficient of determination (R2) of 0.94 and 0.79 respectively, suggesting that the models were adequate to navigate the optimization space. The feasibility of a large-scale biohydrogen fermentation process was studied using the optimized operational setpoints. A semi-pilot scale biohydrogen fermentation process was carried out in 10 L bioreactor and the potential of generating bioelectricity from the process effluents was further assessed using a two-chambered Microbial Fuel Cell (MFC) process. The maximum hydrogen fraction of 46.7% and hydrogen yield of 246.93 ml H2/g TVS were obtained from the semi-pilot process. The maximum electrical power and current densities of 0.21 W/m2 and 0.74 A/m2 respectively were recorded at 500 Ω and the chemical oxygen demand (COD) removal efficiency of 50.1% was achieved from the MFC process. This study has highlighted the feasibility of applying agricultural and municipal wastes for large-scale microbial production of hydrogen, with a simultaneous generation of bioelectricity from the process effluents. Furthermore, the potential of generating an economical feasible biohydrogen production process from these waste materials was demonstrated in this work. Keywords: Biohydrogen production, Organic Fraction of Solid Municipal Waste (OFSMW), Modelling and optimization, Fermentation process, Renewable energy, Bioenergy / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2013.

Abstract available in PDF file. / Thesis (M.Sc.)-University of KwaZulu-Natal, Durban, 2012.

Orthophosphate removal from wastewater in a subsurface-flow constructed wetland (SFCW) was studied during the fall of 1995. The SFCW consisted of a cell with an impervious liner, filled with 1.27 to 3.81 cm river rock and planted with common reeds (Phragmites australis) and soft-stem bulrushes (Scirpus validus). Municipal wastewater was pumped through the wetland and comparisons were made between inlet and outlet orthophosphate concentrations. Comparisons were also made at locations within the cell. A mean orthophosphate reduction of 62% with a range of 91% to 32% was observed for the duration of the study. An ANOVA test showed a significant reduction of orthophosphate in the wetland cell during the study. A regression analysis indicated that inlet wastewater temperatures played a significant role in orthophosphate removal, while outlet wastewater and ambient air temperature were not a significant factor.Keywords: Subsurface-Flow Constructed Wetlands, Orthophosphate, Municipal Wastewater, Water Chemistry, Nutrients. / Department of Natural Resources and Environmental Management

The research project was initiated to refine the knowledge available on the treatment of rural municipal wastewater by constructed wetlands. Field and laboratory studies were conducted to determine the treatment capacity of a constructed wetland system and to determine a substrate that would be most efficient as a phosphorus adsorption filter. The wetland system consisted of three free-water surface cells, three blast furnace slag filters and a vegetated filter strip, treating municipal wastewater. Bimonthly water samples at the inlet and outlet of each component of the wetland system were analysed for biochemical oxygen demand, nitrate and nitrite, ammonia and ammonium, total Kjeldahl nitrogen, total suspended solids, total phosphorus, ortho-phosphate, fecal coliforms and E. coli. Phosphorus and nitrogen concentrations were determined in the sediment, plant tissue and water column of the free-surface wetland cells. The free-surface wetland cells achieved removals as follows: ammonia and ammonium (52%), total Kjeldahl nitrogen (37%), total suspended solids (93%), total phosphorus (90%) and ortho-phosphate (82%). The vegetated filter strip achieved removals as follows: ammonia and ammonium (28%), total Kjeldahl nitrogen (11%), total suspended solids (22%), total phosphorus (5%) and ortho-phosphate (0%). The slag filters reduced total phosphorus by more than 99%. Phosphorus adsorption measurements were conducted on slag, calcite and shale. Slag was found to be the most effective at removing phosphate.

This thesis is concerned with the use of natural waste materials, specifically used tea leaves and coffee grounds, as adsorbents for the removal of trace metals from water, e.g. in waste water treatment. Trace metals such as lead, mercury, zinc, copper, nickel and cadmium are released to the environment in waste water as a result of human activities, and generate concern because of their potential toxicity, persistence and tendency to become concentrated in food chains. While there are many conventional methods for removing these metals from waste water, such as chemical precipitation, ion exchange, membrane technologies and electrochemical treatment, these processes can be expensive. Thus in recent years there has been increasing interest in low cost adsorbent materials as alternative adsorbents, particularly waste natural products such as rice hulls and spent coffee grounds. Most of the research reported in this thesis has been conducted with spent coffee grounds, both grounds produced by leaching of commercial ground coffee and spent grounds obtained from the manufacture of instant coffee. However, some preliminary work was also conducted using spent tea leaves. In the initial work, the adsorption of the metal ions Cu�⁺, Zn�⁺, Cd�⁺ and Pb�⁺ by these adsorbents was investigated using batch adsorption methods to determine the effects of pH, metal ion concentration, adsorbent concentration and other factors such as temperature and metal ion competition. The competitive effects of soluble material leached from the adsorbents that also bind metal ions were studied. The adsorption of the metal ions was found to follow the Langmuir adsorption isotherm. However, the maximum adsorption density was found to depend on the concentration of coffee adsorbent present. Further investigation indicated that this was a result of flocculation of the coffee solids, which acted to reduce the available surface area and thus the maximum density of adsorption sites. This was confirmed using a dispersant to break up the flocculated solids. Column adsorption studies showed that metal ions adsorbed by coffee grounds could be quantitatively leached in dilute acid solution without any loss of subsequent adsorption properties, thus promoting efficient re-use of the column for many adsorption cycles. The adsorbent was also found to be largely unaffected by biological degradation. A prototype waste water treatment column was used to treat tap water samples, with and without known additions of metal ions. The results showed that the grounds efficiently adsorbed trace metal contaminants at levels as low as [mu]g L⁻�. The acid base chemistry of both tea leaves and coffee grounds, and the soluble materials leached from the fresh tea and coffee, were studied using potentiometric titration. In addition the stoichiometry of H⁺ released during metal ion adsorption was also investigated. The latter results indicated that the stoichiometry of metal ion adsorption is not simple, i.e. it probably involves more than one type of adsorption site. The results of this thesis suggest that the use of waste coffee grounds shows considerable promise for the treatment of waste water containing trace metals, and provides an alternative commercial use for such exhausted coffee materials.