A dissertation submitted to the Faculty of Science, University of the Witwatersrand, in fulfilment of the requirements for the degree of Master of Science in, Microbiology and Biotechnology 2016. / Heavy metal contamination, of natural water resources, resulting from the large amounts of toxic waste generated by industrial practices is of great environmental concern. Lead (Pb) in particular is one of the most toxic heavy metals that leads to several health deficiencies upon human exposure. The reduction of heavy metals like Pb to acceptable levels in the water therefore becomes critical for potable and agricultural use. Removal of heavy metals by conventional methods is expensive and results in secondary pollution. Bioremediation, a process that passively removes heavy metals from solution through microbial biosorption, is a much sought after alternative because it is more eco-friendly and cost-effective. Micrococcus luteus and Paenibacillus castaneae are two bacterial species reported to be highly resistant to Pb making them favourable as metal biosorbents. The present study aimed to further characterise these species as biosorbents by evaluating the influence of environmental conditions on their rate of biosorption of Pb. Each bacterial isolate was heat-killed and exposed to 0.5 mM (150 mg/L) Pb and the maximal rate of metal uptake calculated when the pH, temperature and biomass concentration were varied. Additionally, the initial metal concentration was increased from 0.005 to 1.25 mM to determine its effect on Pb uptake by each species. The influence of competing cations (Ni2+, Co2+, Mn2+ and Zn2+) on the rate of Pb uptake by each isolate was also established. Both bacterial isolates resulted in the biosorption of at least 50% of 0.5 mM Pb ions when used at a pH of 7, temperature of 25 oC, and a biomass concentration of 2 g/L. The rate of metal uptake for M. luteus at the above mentioned parameters was found to be 24.51 mg/g biomass, while the rate of metal uptake for P. castaneae was 15.63 mg/g biomass. These findings indicated that M. luteus took up more Pb at a faster rate in comparison to P. castaneae. The present study furthermore elucidated that as the metal concentration of Pb was increased, the amount of Pb biosorbed by M. luteus decreased from 84.76% to 46.10%. Similarly, P. castaneae yielded 81.39% biosorption from 0.005 mM Pb but only 34.29% of Pb was taken up when the concentration was increased to 1.25 mM. When the bacteria were exposed to various competing cations an increase in the rate of Pb biosorption was observed for P. castaneae while the opposite effect was noted for M. luteus. Findings from this study show that under high metal concentrations, both M. luteus and P. castaneae are capable of significantly reducing the level of Pb from pure solution. The results warrant further treatment of several industrial effluents using these biosorbents for subsequent application in wastewater treatment. / MT2017

The potential of non-viable Azalia filiculaides for the removal of Cu and Ni from aqueous solutions and the possibility of scaling up existing lab scale Azalia column systems was investigated. The effects of factors such as metal starting concentration, pH and two metals in solution on the removal of Ni and Cu from aqueous solution by dried and crushed Azalia biomass were studied in batch systems. Aqueous solutions of Ni with starting concentrations between 1000 and 2000J.lmolll gave the most efficient Ni removal by Azalla biomass. For Cu the optimum starting concentration for adsorption was 50J.lmol/l. The adsorption capacity of both eu and Ni increased as the starting pH of the sorption media increased. The optimum pH for Ni adsorption was found at pH 7 and for Cu, at pH 5. - Awlla biomass had a higher. maximum binding capacity (qrnax) for Cu than for Ni at pH 5. The removal of both Cu allct Ni showed little or no variation with the presence another metal in solution. Kinetic studies show that both Cu and Ni adsorbed rapidly onto the Azalia biomass. The removal of Cu and Ni from aqueous solutions using non-viable Azalia biomass was investigated in a lab scale fixed-bed column and an upscaled 4L column system. The nonviable Azalla filiculaides biomass when dried and used in a column for adsorption of Cu and Ni showed good physical stability under many different conditions. Preparation of the biomass before it could be used in the columns was very simple and did not involve any significant pretreatment steps. Prolonged exposure to UV light decreases Azalia biomass capacity for Ni and Cu adsorption. Column adsorption of Cu and Ni from aqueous solutions was successfully upscaled approximately 100 times. Relative to the lab scale column, the 4L column performed better for the uptake of Cu and Ni per gram of biomass. The larger column was also able to operate at relatively higher flow rates. The biomass showed good reusability with little change in the amount of Ni adsorbed in 10 consecutive cycles. Electron micrographs showecf little or no change in the physical structure and integrity of the Azolla biomass after exposure to mineral acids, Ni solution and high flow rates over 10 consecutive adsorption and desorption cycles. As much as 80% Ni and 70 % Cu was recovered when desorption profiles were generated using O.lMHCI as a desorption agent. The 4L column system was also tested using a highly concen~rat:~ Ni plating bath solution.(Nicrolyte 1). Only 18 % of the Ni could be removed from the expended Nicrolyte 1 pla~Jng solution after treating only 25L, indicating that Azolla biomass is more suited for removal of metals from more dilute industrial effluents.

Thesis (MTech (Chemical Engineering)--Cape Technikon, Cape Town, 1998 / Ion exchange resins are widely used to remove or concentrate heavy metals from aqueous solutions or slurries.This thesis attempts to properly evaluate the interaction between ion exchange resins and heavy metals at trace metal concentrations.The durability of the resins and their effectiveness in real slurries were also investigated. In this study, a chelating resin, as well as a cation, and anion exchange resin was contacted with aqueous solutions of heavy metals in both free and complexed form. Zinc, nickel and copper cyanide complexes were adsorbed onto the anion exchange resin, while the chelating and cation exchange resins were contacted with zinc and nickel nitrates, and cupric sulphate. All the tests were conducted in batch stirred tank reactors. All the metal cyanide complexes behaved in a similar manner when contacted with the anion exchange resins. These tests were p~rf0nned under variations in temperature, stirring speed, pH., ionic strength and . initial metal 90E~entrations. Fitting of a dual resistance model to the profiles for thetlptllk:e" of the complexes, show that both film diffusion and intraparticle diffusion rates were improved with an increase in temperature, and that film diffusion rates improved with an increase in stirring speed. A high ionic strength negatively affected equilibrium loading as well as diffusional rates.It was found that at these low concentrations, the diffusional rates improves with a decrease in the external metal concentration. A comparative study involving the chelating and cation·exchange resins were performed, during which the resins were contacted with the metals in free fonn. It was found that at high metal concentrations, the chelating resin induced a rate limiting effect, but at trace concentrations, this effect is virtually negated. Whereas the cation exchange resin exhibited little selectivity in adsorbing the metals, it was found that the chelating resin prefers the metals in the eu > Ni > Zn. The chelating resin proved to be no less durable then the cation exchange resin, and both slightly lost their ability to adsorb the metal cations as a result of the effects of an inert coarse sand slurry.Tests performed with a real ore leachate, showed the cation exchange resin to be efficient at a low pH , but also relatively non selective, since the adsorption of copper from the leachate was greatly reduced due to the presence of other heavy metals.

M.Tech. (Extraction Metallurgy) / For many decades, carbon nanotubes (CNTs) have been used as adsorbents for the removal of pollutants from wastewaters because of their unique properties such as inert surface, resistance to acid and base environment, rigidity and strength. Herein is a report of application of functionalised CNTs on the adsorption of metal ions from aqueous solutions and mine leachates. The CNTs were first synthesised in-house, purified, functionalised and characterised with various characterisation techniques: FTIR (Fourier Transform Infrared), SEM (Scanning Electron Microscopy, TEM (Transmission Electron Microscopy), EDS (Energy Dispersive Spectroscopy), Raman Spectroscopy, TGA (Thermal Gravimetric Analysis) and BET (Brunauer-Emmet-Teller). Together, these techniques gave substantiation for structure, surface and chemical modification of the synthesised moieties. After characterisation, the functional groups were attached to the walls of the tubes and this implies successful functionalisation...

M.Sc. (Chemistry) / Polymer nanocomposites are a new class of composites in which at least one dimension of the particles dispersed in the polymer matrix is in the nanometer range. Recently, different types of zeolite minerals, either natural (Clinoptilolite, chabazite, modernite) or synthetic (A-type, X-type, Y-type) are being employed as particulate fillers into the polymer matrix. Owing to their unique ion exchange phenomenon, zeolites have been widely studied as heavy metal adsorbents, but very few researchers have focused on the sorption of heavy metal ions on zeolite-filled polymer nanocomposites...

The study was conducted to investigate the effects of: (i) varying the level of supply of the heavy metals iron, manganese, copper and zinc. (ii) varying the ratio between different pairs of metals in the medium. In particular the iron:manganese and zinc:copper ratios were studied. Initially the two fungi Aspergillus niger van Tieghem, (variety and strain) and Penicillium notatum Westling, were used. Penicillium notatum Westling was subsequently discarded, in favour of Penicillium glancum Link, because it did not sporulate freely in liquid culture. The fungi were grown in controlled nutrient solutions, and during the course of tho growth and development, the form and sporulation of the felts was noted. After a period of growth, the felts were removed, dried and weighed. The pH of the liquor was measured. the results were studied to determine the effect of varying levels of supply of the heavy metals, and of the varying heavy metal ratios in the culture solution. In the investigation of the effect of varying the level of supply of individual heavy metals, optimum concentrations were demonstrated for copper and manganese. Increaning the concentration of pairs of heavy metals cimultaneously was found to influence the appearance and degree of symptoms of toxicity. Cultures of Aspergillus and Penicillium were found to be able to tolerate concentrations of copper, in particular, considerably greater than the observed optima, when zinc was present in equal concentration. Citric acid, and subsequently ethylene- diaminetetra-acetic acid, were used as chelating agents, in order to prevent the precipitation of the metals in the culture solution during autoclaving. It was found that the use of chelating agents markedly reduced symptoms of toxicity. There was no conclusive evidence that the iron:manganese ratio in the culture medium was an important factor in the growth and development of cultures of Aspergillus and Penicillium. However there is considerable evidence that in cultures of Penicillium, the zinc:copper ratio in the medium is of some importance in the determination of the dry weight yield trends. This effect was not demonstrated in cultures of Aspergillus.

This study investigated the possible use of algal extracellular polysaccharide as a biosorbent for removal of heavy metals (copper and lead) from aqueous solutions as a means of bioremediation for metal containing effluents. This biopolymer has good biosorbent properties and a potential to provide a cost effective, selective and efficient purification system. A variety of environmental conditions induce the production of extracellular polysaccharides in algae. The production of exopolysaccharides by Dunaliella cultures was induced by nitrogen deficient conditions. A high ratio of carbon to nitrogen source considerably enhanced the polysaccharide release. Purified extracellular polysaccharide samples exhibited a monosaccharide composition consisting of the following sugars: xylose, arabinose, 2-0-methyl mannose, mannose, glucose and galactose. The relative abundance (%) of these sugars were calculated relative to xylose. The major sugar constituent was 2-0-methyl mannose, which was present at approximately 160% relative to xylose. The percentage relative abundance of other sugars was as follows: 18.8; 86.8; 85.3 and 22.3% for arabinose; mannose; glucose and galactose respectively. The identity of the various constituents were confirmed by mass spectrometry. The ability of Dunaliella exopolysaccharides to accumulate metals was investigated. The following parameters were studied because they affect metal uptake: solution pH, biomass concentration, temperature, time and metal concentration. The uptake of both copper and lead were pH dependent. However, metal uptake was not significantly affected by temperature. Kinetic studies showed that Dunaliella extracellular polysaccharides exhibit good bioremediation properties. Metal uptake was rapid. In addition, the exopolysaccharide has good metal binding capacity with an uptake capacity for lead of 80 mg/g from a solution containing initial lead concentration of approximately 40 mg/l. Competition studies revealed that the presence of a second metal in solution inhibits uptake of the other metal compared to uptake in single metal solution of that particular metal. The presence of lead inhibited the uptake of copper from approximately 65% in single metal solution to 10% in binary metal solution. The presence of copper also inhibited lead uptake, though not to the same extent. Higher concentrations of lead could not completely prevent removal of copper from solution and visa versa. The same was true for lead which could not be displaced by a four-fold concentration of copper. Instead, a certain percentage of copper was always removed showing that lead did not compete with copper for these binding sites. In conclusion it appears that, copper and lead bind to different sites on Dunaliella exopolysaccharides and that they exhibit selective or preferential removal of lead.

The removal of lead from aqueous solution and lead-acid battery manufacturing waste-water by the non-viable biomass of the water fern Azolla filiculoides was investigated in both batch and column reactors. The maximum lead uptake by the Azolla biomass at a pH value of approximately 5, was found to be 100 mg lead/g biomass from aqueous solution. Lead removal varied from 30% of the initial lead concentration at pH 1.5 to approximately 95% at pH values of 3.5 and 5.6. Lead removal from aqueous solution decreased to 30% of the initial lead concentration if the lead concentration was initially over 400 mg/l. At initial lead concentrations of less than 400 mg/l, percentage lead removal was found to be over 90% of the initial lead concentration. Lead removal remained at approximately 90% between 10°C and 50°C. Biomass concentration (4-8 mg/l) had little effect on lead removal. The presence of iron (Fe) and lead, copper (Cu) and lead or all three metal ions in solution at varying ratios to each other did not appear to have any significant effect on lead removal. Percentage lead, copper and iron removal from aqueous solution was 80-95, 45-50 and 65-75% respectively for the different multiple-metal solutions studied. No break-through points were observed for lead removal from aqueous solutions in column reactors, with initial lead concentrations of less than 100 mg/l at varying flow rates of 2, 5 and 10 ml/min. This suggested that flow rate, and therefore retention time, had little effect on percentage lead removal from aqueous solution, which was more that 95%, at low initial lead concentrations (less than 100 mg/l). At initial lead concentrations of 200 mg/l or more, an increase in flow rate, which equates to a decrease in column retention time, resulted in break-through points occurring earlier in the column run. Percentage lead removal values, from lead-acid battery efiluent in column systems, of over 95% were achieved. Desorption of approximately 30% and 40% of bound lead was achieved, with 0.5 M HNO₃ in a volume of 50 ml, from two lead-acid battery. Repeated adsorption and desorption of lead by the Azalia biomass over 10 cycles did not result in any decrease in the percentage lead removal from effluent, which strongly suggested that the Azalla biomass could be re-used a number of times without deterioration in its physical integrity, or lead removal capacity. No evidence of deterioration in the Azolla biomass's physical integrity after 10 successive adsorption and desorption procedures was observed using scanning electron microscopy. The Azolla filiculoides biomass was, therefore, found to be able to effectively remove lead from aqueous solution and lead-acid battery effluent repeatedly, with no observed reduction in it's uptake capacity or physical integrity.

M.Sc., Faculty of Science, University of the Witwatersrand, 2011 / Contamination of water bodies by heavy metals and metalloids is an established problem and several studies have been conducted to deal with it. South Africa is amongst those countries whose water systems are most affected as a result of intensive mining activities. This research was dedicated to the development of insoluble chelating polymers for use as adsorbents to abstract heavy metal ions from mining and industrial wastewater. Branched polyethylenimine (PEI), well known for its metal chelating potential, was cross linked by epichlorohydrin in order to convert it into a water-insoluble form. The water-insoluble property gives the advantage of being used in situ and a possibility of regeneration and re-use, making it a more feasible and cost-effective method. Its surface was also modified for selective removal of specifically-targeted heavy metal and metalloid ions. The binding affinity of the synthesized materials to heavy metal and metalloid ions has been determined as well as their ability to be regenerated for reuse. These processes demonstrated that cross-linked polyethylenimine (CPEI) exhibited good complexation ability with high affinity to Cr and some divalent metal ions such as Fe, Zn, and Ni. On the other hand, it showed very poor ability to bind oxo-anions such as SeO32- and AsO2- which has been attributed to the unavailability of suitable functional groups to interact with these ions. The observed order of complexation was: Cr > Zn> Fe >> Ni > Mn > Pb >> As > U > Se. The phosphonated polyethylenimine (PCPEI) showed high selectivity for As, Mn and uranyl ions. The observed order of removal was: U > Mn> Ni > Zn > As >> Cr > Pb > Fe >> Hg > Se; whereas the suffocated polyethylenimine (SCPEI) exhibited high affinity to Se, and Hg. The observed order of adsorption was: Hg > Se >> U > Zn >Pb > Ni >> As > Cr > Fe. v The adsorption behaviour of these polymeric materials involved more than one mechanism such as complexation, normal surface charge exchange, and anion replacement and all these mechanisms are governed by the functional groups. The nitrogen atom on the chelating group (-NH) in the cross-linked polyethylenimine; the phosphorus atom on the chelating group (-PO3H2) in phosphonated cross-linked polyethylenimine; and sulphur atom on the chelating group (-SO3H) in suffocated cross-linked polyethylenimine act as Lewis bases and donate electrons to metal cations which are considered Lewis acids. The existence of the chelating groups in SCPEI and PCPEI facilitate the removal of oxo-anions through anion replacement since they exist as bases in solution and hence cannot be electron acceptors. Thus, the expected mechanism is the normal anion replacement. This mechanism can explain the high removal of Se by SCPEI since Se has similar chemical behaviour as sulphur and are in the same group in the periodic table. As such they can easily replace each other. Sulphur is released from the polymer into the solution by replacing the selenium ions in the polymer. Similar behaviour occurs between phosphorus in PCPEI and arsenic ions as As and P belong to the same group in the periodic table and hence have similarities in their chemical behaviour. The Langmuir and Freundlich isotherm models were used to interpret the adsorption nature of the metal ions onto synthesized polymers. The Freundlich isotherm was found to best fit and describe the experimental data describing the adsorption process of metal and metalloid ions onto the synthesized polymeric materials The kinetic rates were modelled using the pseudo first-order equation and pseudo second-order equation. The pseudo second-order equation was found to explain the adsorption kinetics most effectively implying chemisorption. vi The thermodynamic study of the adsorption of metals and metalloids by the synthesized CPEI, PCPEI and SCPEI resulted in high activation energies > 41 KJ mol-1 which confirm chemisorption as a mechanism of interaction between adsorbate and adsorbent. So far, the developed polymeric materials showed good results and have potential to be applied successfully for remediation of heavy metal-polluted waters, and they have potential for use in filter systems for household use in communities that use borehole water impacted by mining and industrial waste waters. The desorbed metals can be of use to metal processing industries.

The research presented in this thesis entailed an investigation of heavy metal uptake by Palmaria palmata, a red marine alga. The alga was dry and organically certified. The heavy metals of interest were those commonly found in the wastewaters of the printed wiring board industry, namely Cu2+Pb2+, Cd2+, Zn 2+ and Ni2+. The ultimate objective of the work was to determine whether or not the factors expected to influence the metal uptake to the greatest extent could be optimized within functional ranges, leading eventually to process design (beyond the scope of this thesis). These factors were pH, temperature, initial concentration of metal in aqueous solution, and contact time. A number of preliminary experiments were performed to establish a basis for the design of the optimization studies. / Although it was not possible to adequately define optimal regions of operation for the biosorption of heavy metals by Palmaria palmata , general trends were elucidated, and the limitations of the methodology used were clarified. (Abstract shortened by UMI.)