A thesis submitted to the Faculty of Science, University of the Witwatersrand in fulfilment of the requirements for the degree of Doctor of Philosophy. November 2016. / This research was directed towards the environmental monitoring and assessment of the most used non-steroidal anti-inflammatory drugs (NSAIDs) which are naproxen, ibuprofen and diclofenac. The work involved the development and application of sensitive techniques for the quantification of naproxen, ibuprofen and diclofenac in the South African aquatic environment. Based on this information, a multi-templates molecularly imprinted polymer (MIP) was synthesized and applied alongside the commercial available sorbent (Oasis MAX) in the solid-phase extraction (SPE) of target compounds from water samples. The extracted compounds were then quantified using high performance liquid chromatography (HPLC). MIP was synthesized by applying a bulk polymerization approach at 70 ͦ C where all target compounds were used as multi-templates. Other reagents used in synthesis were 2-vinyl pyridine, 1,1’-azobis-(cyclohexanecarbonitrile), ethylene glycol dimethacrylate and toluene as functional monomer, initiator, cross-linker and porogenic solvent, respectively. Synthesis of a non-imprinted polymer (NIP) under similar reaction conditions as MIP was carried out with the omission of templates. Techniques employed in characterization of MIP and NIP were Fourier transform infrared spectroscopy (FTIR), Brunauer, Emmett and Teller (BET) method, CHNS analyzer, zeta potential, cross-polarization/magic angle spinning nuclear magnetic resonance spectroscopy, thermogravimetric analysis, differential scanning calorimetry and x-ray diffraction. Monomer-template interactions were investigated using molecular dynamics. The performance of the MIP was evaluated based on its ability to selectively extract target compounds in organic (acetonitrile, acetone, chloroform and toluene) and aqueous media. The extraction capacity of the MIP in organic solvents for naproxen, ibuprofen and diclofenac increased from high polarity to low polarity solvents. In a low polarity solvent (toluene), the extraction capacity achieved for naproxen, ibuprofen and diclofenac were 14.4, 11.0 and 14.0 mg/g, respectively. In this case, the selectivity of the MIP where gemfibrozil was employed as the competing species was evident. Selectivity of the MIP collapsed during the adsorption of naproxen, ibuprofen and diclofenac from water using gemfibrozil and fenoprofen as competitors. This resulted in high extraction efficiencies for target compounds and competitors, however, both gemfibrozil and fenoprofen were easily desorbed from the MIP using weak organic solvent due to lack of molecular recognition. During the binding sites characterization, the best fit of pseudo-second-order implied a chemisorption of all target compounds onto MIP sorbent. The data also fitted well in Langmuir isotherm which meant that the adsorption of target pharmaceuticals occurred on the homogeneous binding sites of the MIP. Optimized adsorption conditions in water such as MIP amount of 50 mg, extraction time of 10 min, sample pH of 2.5 and sample volume of 10 mL were applied for the selective adsorption of naproxen, ibuprofen and diclofenac in contaminated wastewater and river water. In WWTP influent, naproxen recovery was 38%, whereas ibuprofen and diclofenac were 69% and 87%, respectively. MIP was further used as a selective adsorbent in solid-phase extraction (SPE) of three drugs from environmental samples. The selectivity of the MIP in environmental samples was compared to that of the commercially available Oasis MAX sorbent. The application of molecularly imprinted solid-phase extraction (MISPE) reduced matrix effects and improved the sensitivity of the analytical method. In this case, the detection limits for naproxen, ibuprofen and diclofenac were 0.2, 1 and 0.6 μg/L, respectively. When deionized water was spiked with 5 and 50 μg/L of target compounds, recoveries greater than 80% were obtained. Thereafter, the developed MISPE was applied for selected acidic drugs from environmental samples. Environmental samples were collected from urban (Durban) and semi-urban/rural areas (Ladysmith) of KwaZulu-Natal Province in South Africa. The most abundant compound in the environment was ibuprofen. In river water samples from Durban, the maximum concentrations found for naproxen, ibuprofen and diclofenac were 6.8, 19 and 9.7 μg/L, respectively. The maximum amounts found for the same drugs in Ladysmith river samples were generally lower with naproxen, ibuprofen and diclofenac detected at 2.8, 6.7 and 2.6 μg/L, respectively. The same trend was observed in wastewater. Further work on the monitoring of acidic compounds in wastewater was conducted using Oasis MAX as the SPE sorbent prior to HPLC analysis. All target compounds were detected in Kingsburg and Umbilo WWTPs located in Durban surroundings. The influent and effluent concentrations detected were in the ranges of 6.4 to 69 μg/L and 0.6 to 4.2 μg/L, respectively. Further to this, the removal efficiency of the target compounds during the WWTP process in Kingsburg and Umbilo was in the range of 69 to 97%. The extent of pollution in the environment was further assessed by the monitoring of ketoprofen and triclosan in wastewater and river water using SPE with Oasis HLB sorbent and HPLC. Traces of both compounds ranging from 1.2 to 9.0 μg/L were detected in wastewater. The maximum concentrations found in river water were 2.0 and 0.9 μg/L for ketprofen and triclosan, respectively. Overall, the analytical methods implemented in this work were highly accurate, precise and sensitive. The synthesized MIP was highly selective and its application in environmental studies led to the development of a less expensive analytical method. This work also gives the overview of the extent of water pollution caused by acidic pharmaceuticals in various water matrices. / MT2017

Submitted in fulfillment for the requirements for the Degree of Doctor of Technology: Biotechnology, Durban University of Technology, 2012. / Nitrification is a crucial step in biological nutrient removal (BNR) processes, mostly carried out by a group of nitrifying bacteria which includes ammonia-oxidising bacteria (AOB) and nitrite-oxidising bacteria (NOB). Nitrification failure has proven to be a common operational problem in full-scale wastewater treatment plants (WWTP) since nitrifying bacteria are very sensitive to sudden changes in environmental or plant operating conditions. The current investigation was carried out to advance our understanding of the distribution of nitrifying bacterial populations and their performance at three different BNR plants in KwaZulu-Natal, South Africa. The latest molecular techniques such as fluorescent in situ hybridisation (FISH)-confocal scanning laser microscopy (CSLM), polymerase chain reaction (PCR) and real-time quantitative PCR (Q-PCR) were applied to detect and quantify nitrifying bacteria. When using FISH to target the nitrifying population, it necessitated optimising pre-treatment protocols of the samples to improve accuracy during quantification. Sonication was found to be the superior method of dispersion based on the least disruption of nitrifier cell integrity, irrespective of the sludge type. The effect of plant configurations and wastewater characteristics on the distribution of the nitrifying bacterial population and subsequently on the nitrification performance was evaluated using FISH and PCR. FISH results revealed the dominance of Nitrosomonas (AOB), Nitrobacter (NOB) and Nitrospira (NOB) for all BNR plants. The 16S rRNA analysis of PCR products using genus-specific primers, revealed the presence of more than one species of the same group at these plants. Nitrosomonas spp. including Nitrosomonas halophila, Nitrosomonas eutropha, Nitrosomonas europaea, Nitrosomonas aestuarii and an unidentified Nitrosomonas spp. were found to dominate among the AOB and Nitrobacter vulgaris, Nitrobacter alkalicus, Nitrobacter hamburgensis and an unidentified Nitrobacter spp. were the dominant species for NOB. Among these species, Nitrosomonas aestuarii, Nitrosomonas europaea, Nitrobacter hamburgensis were detected only from the industrial wastewater samples. The efficiency of two commonly used techniques viz., FISH and Q-PCR for the detection of nitrifiers from WWTP were also studied and compared, specifically targeting Nitrobacter sp. Even though there were slight variations in the quantification results, changes in the Nitrobacter community at these plants were consistent for both FISH and Q-PCR results. Both techniques have their own limitations and advantages. This study has helped to add to the platform of understanding the distribution and activity of nitrifying bacteria by correlating population dynamics with the operational parameters at full-scale level. The observations made in this study will assist researchers and engineers to minimise future nitrification failure at full-scale BNR plants. This study also confirmed the highly complex activities of wastewater treatment processes, which is dependant on a number of factors. Specific AOB or NOB predominant in wastewater rather suggests that the wastewater type and characteristics may contribute to significantly different microbial environments. Among the AOB, Nitrosomonas dominated at all BNR plants throughout the study period and for NOB both Nitrobacter and Nitrospira were found in significant numbers but their dominance varied across the plants. These dissimilar, distinct distribution patterns could be attributed to their environment which in turn impacted on the nitrification performance of the system. It was also noted that the co-existence of more than one group of these communities at the same plant could help the plant escape complete functional failures such as nitrification, due to sudden changes in temperature and substrate concentrations, as this function can be performed by different groups. Although it would have been meritorious to conduct a nitrogen balance in this study, this was not possible since the research focused on full-scale systems. / National Research Foundation / D

Submitted in fulfilment of the requirements for the Degree of Master of Engineering, Durban University of Technology, 2017. / In this research, the woven fibre microfiltration (WFMF) fabric which is produced locally in South Africa is used as a membrane material. It is cheaper in price in comparison with the current commercial membrane materials that are in use. The WFMF is also more robust when compared with the commercial membrane materials thus is able to withstand harsh working conditions. From previous studies on the WFMF, it has been shown that it can be used as a membrane material without any compromise to permeate quality. This research seeks to optimize the working conditions of this membrane material (WFMF) with an aim of achieving lower running costs and better anti fouling strategies in comparison to the commercial MBRs. The objectives and aims of this research was to come up with a MBR system whose running cost is lower than that for the commercial systems, which can be adapted for use in any environment, especially in the hardship regions where its robustness would be an added advantage. The performance of the WFMF submerged MBR was also optimised including antifouling operating regimes. This study was done in a pilot plant that was set up at Veolia wastewater treatment plant, Durban Metro Southern Works. The feed water for the pilot plant was pumped from the return activated sludge mixing chamber by means of a submersible pump. The MLSS concentration of the feed water was about 12 g/l. The various investigations that were conducted in the course of this research included the effect of spacing between membrane modules, relaxation steps and frequencies, evaluation of aeration rates and evaluation of coarse vs. fine bubbles which were all aimed at optimizing the performance of the immersed WFMF MBR. The permeate was checked for turbidity and COD levels to ensure that they were within the accepted water standards. From the experiments it is shown that the critical flux increased with an increase in aeration rate which is in concurrence with the literature and a starting flux of 30 LMH was chosen for the running of the pilot plant for the various experimental runs to be carried out. For the pipe diffuser height effect experimental run, the best results were achieved at a height of 5 cm below the membrane modules and the use of a pipe diffuser gave better results than the use of a disc diffuser. For the membrane module spacing effect the best results were obtained at the smallest possible width i.e. 3.5 mm. The best relaxation step sequence was found to be 9 mins on and 1 min off. COD, turbidity and DO was continuously determined during the course of the experimentation. Further studies should be done on use of the disc diffuser with increased surface area of aeration holes and also hole sizes of smaller diameters to check on its effectiveness as a means of reducing fouling on the membrane surface. / M

M.Sc. (Chemistry) / Prosperity for South Africa depends on the sound management and utilization of many resources, with water playing a crucial role. Located largely in a semi-arid part of the world, South Africa’s water resources are, in global terms, scarce and extremely limited. A key environmental problem facing South Africa is water pollution. This arises from many sources, including mining and industrial effluents, and runoff of biocides, nutrients and pathogens from agricultural lands, urban areas and informal settlements with poor sanitation. The consequences are often severe, including among other impacts, habitat destruction, reduced oxygen levels, fish kills and loss of human life. Inorganic and organic pollutants as well as microbes are the main constituents of the effluent from the domestic, mining, agriculture, metal electroplating, petrochemical and transport industries. The presence of pollutants in environmental systems is of concern because ultimately, they are incorporated into drinking water and various food chains. Therefore, the overall focus of this study was mainly metals analysis in wastewater systems before and after treatment processes so as to establish the efficiency of the treatment processes....

Thesis (M.Sc. (Aquaculture)) -- University of Limpopo, 2014 / Population growth in urban areas is putting pressure on sewage treatment plants. The improper treatment of sewage entering the aquatic ecosystems causes deterioration of the water quality of the receiving water body. The effect of sewage effluent on the Sand River was assessed. Eight sampling sites were selected, site 1 and 2 were upstream of the of the sewage treatment plant along the urbanised area of Polokwane, whilst sites 3, 4, 5, 6, 7 and 8 were downstream. The physicochemical parameters and coliform counts in the water samples were determined. Macroinvertebrate abundances and diversity (H') was determined at the different sites during the dry and rainy season. The water quality status of the Sand River with respect to the South African scoring system (SASS) scores and average score per taxon (ASPT) was determined. A linear regression was performed to test the correlation of the SASS scores with abundance and H'. Heavy metal concentrations in water, sediment, grass (Ishaemum fasciculatum) and fish (Oreochromis mossambicus) at the sites were evaluated. The suitability of the Sand River and surrounding borehole water for irrigation was also determined. Hierarchical average linkage cluster analysis produced two clusters, grouping two sites above the sewage treatment works and six sites downstream of the sewage effluent discharge point. Principal component analysis (PCA) identified total nitrogen, total phosphorus, conductivity and salinity as the major factors contributing to the variability of the Sand River water quality. These factors are strongly associated with the downstream sites. Canonical correspondence analysis (CCA) indicated that Chironomidae family was found on the nitrogen gradient during the dry season. However during the rainy season, Chironomidae was found in the centre of the ordination which indicated that it was ubiquitous. Escherichia coli levels (1463.73 counts/100ml) in the maturation ponds of Polokwane wastewater treatment works could potentially lead to contamination of the Polokwane aquifer. High diversity was recorded at the sites before discharge and the sites further downstream. There was significant correlation (P<0.05) between the SASS scores and macroinvertebrate diversity during the dry season (R2=0.69) and the rainy season (R2=0.77). Fish samples had significantly higher (P<0.05) iron and copper concentrations, while the vi sediment had significantly higher lead concentration (P>0.05). The United States salinity laboratory (USSL) diagram indicated that the sodium hazard (SAR) and alkalinity hazard (conductivity) was low and the Sand River and borehole water was suitable for irrigation. The residual sodium carbonate (RSC) was below 1.24 meq/l, also indicating that both the Sand River and borehole water is still suitable for irrigation. The total phosphorus concentrations fluctuated across the different site. Total nitrogen concentrations showed a gradual decrease downstream from the point of discharge. The coliform levels also showed a gradual decrease downstream. This shows that the river still has a good self-purification capacity.

Waterborne diarrhoeal infections continue to be a major health setback in developing countries, especially in rural areas which lack adequate supply of portable water and sanitation facilities. Globally, waterborne diarrhoeal infections occur with an estimated mortality rate of 10–25 million deaths per year, 95% of which are children under the age of 5 years. The Vibrio species is one of the major groups of enteric pathogens that are responsible for diarrhoeal infections. Many strains of these bacterial species continue to cause epidemics of diarrhoea throughout the world. In this study, the prevalence of Vibrio pathogens in wastewater final effluents was assessed. Wastewater final effluent and discharge point samples were collected monthly between September 2012 and August 2013. All samples were collected aseptically using sterile 1 L Nalgene bottles containing 0.5 ml of sterile sodium thiosulphate solution and transported on ice to the laboratory for analyses within 6 h of collection. The membrane filtration method was used for enumeration of presumptive Vibrio densities on thiosulfate citrate bile salt (TCBS) agar plates. Polymerase chain reaction (PCR) was then used to confirm the identities of the presumptive Vibrio species using the species-specific primers. The confirmed isolates were further subjected to molecular characterization to confirm their respective pathotypes. Presumptive Vibrio densities varied from 0 to 2.11 × 102 cfu/100 ml. Out of 300 confirmed Vibrio isolates; 13.3% (40/300) were Vibrio fluvialis, 22% (66/300) were confirmed to be Vibrio parahaemolyticus, and 24.7% (74/300) proved to be Vibrio vulnificus, and 40% (120/300) were other Vibrio species which were not assessed for in this study. The strains of Vibrio fluvialis were found to exhibit 100% resistance to Polymixin and Tetracycline. However, Gentamicin was active against all the three Vibrio species selected for the purpose of this research. The recovery of Vibrio species in the discharged effluents throughout the sampling period even in adequately disinfected effluents is not acceptable considering the fact that Vibrio is a pathogenic bacterium. The findings of this study underline the need for constant monitoring of the microbiological qualities of discharged effluents and might also be suggestive for a review of the disinfection methods used at the treatment works.

Integrated Algae Pond Systems (IAPS) are a derivation of the Oswald designed Algal Integrated Wastewater Pond Systems (AIWPS®) and combine the use of anaerobic and aerobic bioprocesses to effect wastewater treatment. IAPS technology was introduced to South Africa in 1996 and a pilot plant designed and commissioned at the Belmont Valley WWTW in Grahamstown. The system has been in continual use since implementation and affords a secondarily treated water for reclamation according to its design specifications which most closely resemble those of the AIWPS® Advanced Secondary Process developed by Oswald. As a consequence, and as might be expected, while the technology performed well and delivered a final effluent superior to most pond systems deployed in South Africa it was unable to meet The Department of Water Affairs General Standard for nutrient removal and effluent discharge. The work described in this thesis involved the design, construction, and evaluation of several tertiary treatment units (TTU') for incorporation into the IAPS process design. Included were; Maturation Ponds (MP), Slow Sand Filter (SSF) and Rock Filters (RF). Three MP's were constructed in series with a 12 day retention time and operated in parallel with a two-layered SSF and a three-stage RF. Water quality of the effluent emerging from each of these TTU's was monitored over a 10 month period. Significant decreases in the chemical oxygen demand (COD), ammonium-N, phosphate-P, nitrate-N, faecal coliforms (FC) and total coliforms (TC) were achieved by these TTU's. On average, throughout the testing period, water quality was within the statutory limit for discharge to a water course that is not a listed water course, with the exception of the total suspended solids (TSS). The RF was determined as the most suitable TTU for commercial use due to production of a better quality water, smaller footprint, lower construction costs and less maintenance required. From the results of this investigation it is concluded that commercial deployment of IAPS for the treatment of municipal sewage requires the inclusion of a suitable TTU. Furthermore, and based on the findings presented, RF appears most appropriate to ensure that quality of the final effluent meets the standard for discharge.

PhD (Environmental Sciences) / Department of Hydrology and Water Resources / See the attached abstract below

Thesis (MPA)--Stellenbosch University, 2014. / The asset management process and strategy has been adopted and implemented within the public sector. Although the National Treasury developed the Capital Asset Management Guidelines to assist with the development and implementation of the asset management system the guidelines have received very little attention over the years. The Auditor General’s findings indicate a lack of proper implementation of the asset management process. The study provides a clear indication of the implementation of the asset management process by most government entities, local government municipalities and the public sector. The outcome is that while most entities have made progress with regards to the implementation, much remains to be done. Most entities implement the asset management process selectively. This does not yield positive and sustainable results and they end up abandoning the whole asset management system. The report provides the process of successful development and implementation of asset management, it also indicate that if there are element that are missed or not properly done the process will not yield constant results or performance and the impact on the organization performance will not be positive. The management of the organization must ensure that asset management is aligned with strategic objectives of the organization and performance measurement must be directly linked with asset management, performance measurement should be used to determine progress and implementation. Service level agreements must be established between all relevant departments including the finance department and there must be agreement on the objectives between all stakeholders. The key to proper implementation is the alignment of the asset management process with the strategic objectives of the organization.

Water is an indispensible and yet a difficult resource to be renewed, thus water scarcity has become one of the major challenges faced worldwide, with the Southern regions of Africa being the most impacted and affected, especially the Eastern Cape Province of South Africa where rural communities depend on receiving waterbodies that are often negatively impacted by wastewater final effluents. This present study was conducted between August and December 2010 to assess the physicochemical and microbial qualities of the final effluents of peri-urban and rural communities based wastewater treatment plants in the Eastern Cape Province. The physicochemical parameters were determined on site and in the laboratory, while bacteriological qualities were determined using culture based techniques. The virological qualities were determined by molecular methods using reverse transcriptase polymerase chain reaction for the target RNA virus and the conventional polymerase chain reaction for the target DNA virus. For both wastewater treatment plants, the physicochemical parameters ranged as follows: chemical oxygen demand (5.95-45 mg/L); total dissolved solids (114.5-187.0 mg/L); salinity (0.12-0.20 psu); temperature (14.2-25.7oC); pH (6.0-7.6); nitrate and nitrites (1.55-6.7 mg/L and 0.023-1.15 mg/L respectively); biological oxygen demand (3.5-7.8 mg/L); turbidity (1.49-6.98 NTU); and chlorine residual (0-2.97 mg/L). Feacal indicator bacteria counts ranged as follows: feacal coliforms (0-1.25×104 cfu/100 ml); total coliforms (0-3.95×104 cfu/100 ml); and enterococci (0-5.0×103 cfu/100 ml). xviii Seventy five percent of the rural community based plant and 80 percent of the peri-urban community based plant were positive for coxsackie A virus, while hepatitis A virus was detected in all the rural community based plant 80 percent of the peri-urban community based plant. This study suggests the need for intervention by appropriate regulatory agencies to ensure regular monitoring of the qualities of final effluents of wastewater treatment facilities in the Eastern Cape Province and ensure compliance to established guidelines.