Dissertations / Theses on the topic 'Biodegradation; Wastewater; Effluent purification'

Thesis (MSc)--University of Stellenbosch, 2003.
ENGLISH ABSTRACT: Large volumes of wastewater are generated annually during the grape harvest season from various processing and cleaning operations at wineries, distilleries and other wine-related industries. South African regulatory bodies dictate that wastewater should have a pH of 5.5 to 7.5 and a chemical oxygen demand (COD) lower than 75 mg/L. However, winery wastewater has a typical pH of 4 to 5 and a COD varying between 2 000 and 12 000 mg/L. Urban wineries channel the wastewater to local sewage treatment facilities and are often heavily fined for exceeding governmental requirements. Rural wineries usually have little or no treatment operations for their wastewater and it is often irrigated onto crops, which may result in environmental pollution and contamination of underground water resources. Various criteria are important in choosing a wastewater treatment system, such as an ecofriendly process that is flexible to withstand various concentration loads and characteristics, requiring low capital and operating costs, minimal personal attention and do not require too much land. In this study, a large variation in COD, pH and chemical composition of the winery wastewater was observed that could be related to varying factors such as the harvest load, operational procedures and grape variety. Wastewater from destemming and pressing operations contained higher concentrations of glucose, fructose and malic acid, which originated from the grape berries. The fermentable sugars (glucose and fructose) contributed to almost half of the COD with a smaller contribution from ethanol and acetic acid. The low pH can be ascribed to relative high concentrations of organic acids in the wastewater. The efficacy of biological treatment systems depends strongly on the ability of microorganisms to form biofilm communities that are able to degrade the organic compounds in the wastewater. Preliminary identification of microorganisms that naturally occur in winery wastewater indicated the presence of various bacterial and yeast species that could be effective in the biological treatment of the wastewater. When evaluated as pure cultures under aerobic conditions, some of the yeast isolates effectively reduced the COD of a synthetic wastewater, whereas the bacterial isolates were ineffective. The most effective yeast isolates were identified as Pichia rhodanensis, Kloeckera apiculata, Candida krusei and Saccharomyces cerevisiae. Our search for cost-effective biological treatment systems led to the evaluation of a Rotating Biological Contactor (RBC) for the treatment of winery wastewater. The RBC was evaluated on a laboratory scale with 10% (v/v) diluted grape juice and inoculated with a mixed microbial community isolated from winery wastewater. The results showed a reduction in the COD that improved with an extended retention time. Evaluation of the RBC on-site at a local winery during the harvest season resulted on average in a 41% decrease in COD and an increase of 0,75 pH units. RFLP analysis of the biofilm communities within the RBC confirmed a population shift in both the bacterial and fungal species during the evaluation period. The most dominant yeast isolates were identified with 18S rDNA sequencing as Saccharomyces cerevisiae, Candida intermedia, Hanseniaspora uvarum and Pichia membranifaciens. All these species are naturally associated with grapes and/or water and with the exception of Hanseniaspora uvarum, they are able to form either simple or elaborate pseudohyphae.
AFRIKAANSE OPSOMMING: Groot hoeveelhede afloopwater word jaarliks gedurende die druiwe-oestyd deur verskeie prosessering- en skoonmaakoperasies deur wynkelders, distilleer- en ander wynverwante industrieë gegenereer. Suid-Afrikaanse beheerliggame vereis dat afloopwater ‘n pH van 5.5 tot 7.5 en ‘n chemiese suurstofbehoefte (COD) van minder as 75 mg/l moet hê. Kelderafloopwater het egter gewoonlik ‘n pH van 4 tot 5 en ‘n COD van 2 000 tot 12 000 mg/L. Stedelike wynkelders voer die afloopwater na ń plaaslike rioolsuiweringsaanleg wat dikwels tot swaar boetes vir oortreding van die wetlike vereistes lei. Plattelandse wynkelders het gewoonlik min of geen behandelingsprosesse vir hul afloopwater nie en gebruik die water dikwels vir gewasbesproeiing, wat tot omgewingsbesoedeling en kontaminasie van ondergrondse waterbronne kan lei. Verskeie kriteria is belangrik in die keuse van ‘n waterbehandelingstelsel, byvoorbeeld ‘n omgewingsvriendelike proses wat verskillende konsentrasieladings en samestellings kan hanteer, ‘n lae kapitaal- en bedryfskoste en minimale persoonlike aandag vereis en min ruimte benodig. Hierdie studie het getoon dat kelderafloopwater ‘n groot variasie in COD, pH en chemiese samestelling het wat met wisselende faktore soos die oeslading, operasionele prosesse en selfs die druifkultivar verband kan hou. Afloopwater van ontstingeling- en parsoperasies het hoër konsentrasies glukose, fruktose en appelsuur wat van die druiwekorrels afkomstig is. Die fermenteerbare suikers (glukose en fruktose) dra tot amper 50% van die COD by, met ‘n kleiner bydrae deur etanol en asynsuur. Die lae pH kan grootliks aan organiese sure in die afloopwater toegeskryf word. Die effektiwiteit van biologiese behandelingstelsels steun sterk op die vermoë van mikroorganismes om biofilmgemeenskappe te vorm wat die organiese verbindings in die afloopwater kan afbreek. Voorlopige identifikasie van mikro-organismes wat natuurlik in wynafloopwater voorkom, het die teenwoordigheid van verskeie bakteriese en gisspesies aangedui. Evaluering van hierdie isolate onder aërobiese toestande het getoon dat sommige van die gis-isolate die COD van ‘n sintetiese afloopwater effektief kon verlaag, terwyl die bakteriese isolate oneffektief was. Die mees effektiewe gis-isolate is as Pichia rhodanensis, Kloeckera apiculata, Candida krusei en Saccharomyces cerevisiae geïdentifiseer. Ons soektog na ‘n koste-effektiewe biologiese behandelingsisteem het tot die evaluering van ‘n ‘Rotating Biological Contactor’ (RBC) vir die behandeling van afloopwater gelei. Die RBC is op laboratoriumskaal met 10% (v/v) verdunde druiwesap geëvalueer en met ‘n gemengde mikrobiese gemeenskap wat uit afloopwater geïsoleer is, innokuleer. Die resultate het ‘n verlaging in die COD getoon wat met ‘n langer retensietyd verbeter het. Evaluering van die RBC by ‘n plaaslike wynkelder gedurende die oesseisoen het gemiddeld ‘n verlaging van 41% in die COD en ‘n verhoging van 0,75 pH eenhede getoon. RPLP analise van die biofilmgemeenskappe in die RBC het ‘n bevolkingsverskuiwing in beide die bakteriese en swamspesies aangetoon. Die mees dominante gisspesies is met 18S rDNA volgordebepaling as Saccharomyces cerevisiae, Candida intermedia, Hanseniaspora uvarum en Pichia membranifaciens geïdentifiseer. Al hierdie spesies word gewoonlik met druiwe en/of water geassosieer en is, met die uitsondering van Hanseniaspora uvarum, in staat om òf eenvoudige òf komplekse pseudohife te vorm.

Thesis (DTech (Chemical Engineering))--Cape Peninsula University of Technology, 2017.
The high rate of industrialisation in most developing countries has brought about challenges of wastewater management especially in the mineral processing industry. Cyanide has been used in base metal extraction processes due to its lixiviant properties thus, its presence in wastewater generated is inevitable. Furthermore, partial and/or the use of unsuitable treatment methods for such wastewater is a potential hazard to both human and the environment. There are several reports on biotechnological treatments of cyanide containing wastewater but few mineral processing industries have adopted this approach. Hence, the thermodynamic study of biodegradation of cyanide containing wastewater was undertaken. The primary aim of this study was to explore the application of bioenergetic models and biological stoichiometry to determine the functionality and thermodynamic requirements for cyanide degrading isolate (Fusarium oxysporum EKT01/02), grown exclusively on Beta vulgaris, for a system designed for the bioremediation of cyanidation wastewater. Chapter 2 reviews some of the applicable thermodynamic parameters such as enthalpy, entropy, heat of combustion, heat capacity, Gibbs energy, including stoichiometry models in relation to their applicability for microbial proliferation in cyanidation wastewater. The chapter places emphasis on the application of agro-industrial waste as a suitable replacement for refined carbon sources for microbial proliferation in bioremediation systems because such systems are environmentally benign. The choice of using agro-industrial waste is due to organic waste properties, i.e. agro-industrial waste is rich in nutrients and is generated in large quantities. Chapter 3 presents the materials and various standardised methods used to address the research gaps identified in chapter 2. For an organism to degrade free cyanide in wastewater, it must be able to survive and perform its primary function in the presence of such a toxicant. Chapter 4 exemplifies both molecular and biochemical characteristics of Fusarium oxysporum EKT01/02 isolated from the rhizosphere of Zea mays contaminated with a cyanide based pesticide. The molecular analyses confirmed the fungal isolate to be Fusarium oxysporum EKT01/02 and the nucleotide sequence of the isolates were deposited with National Centre for Biotechnology Information (NCBI) with accession numbers KU985430 and KU985431. The biochemical analyses revealed a wide substrate utilisation mechanism of the isolate dominated by aminopeptidase including nitrate assimilation capabilities. A preliminary investigation showed free cyanide degradation efficiency of 77.6% (100 mg CN-/L) after 5 days by the isolate. The excess production of extracellular polymeric substance (EPS) was attributed to the isolates’ strive to protect itself from cyanide toxicity.

Abstract Environmental concern and tighter public regulations have created new pressures to reduce effluent loading from kraft pulp bleaching during the past decades. This trend has forced both pulp mills and the related engineering industries to develop and optimise equipment for the pulping processes which fulfils these demands. New pulp washers and washing systems connected with the introduction of new oxygen-based reagents have in particular provided considerably improved tools for reusing process water and thereby essentially reducing effluent discharge or even allowing the implementation of a totally closed water circulation system in kraft pulp bleaching. This study was performed mainly on a laboratory scale at the University of Oulu, but some pilot-scale tests were also performed to confirm the findings of the laboratory-scale evaporation test. Another aim of the pilot-scale tests was to produce enough concentrate for further processing. Conditions were strictly controlled in all the experiments in order to ensure that conclusions could be reached regarding the phenomena examined. The results reported here indicate that the amounts of transition metal ions such as iron, copper or manganese in the process water or water to be recirculated should be kept as low as possible during chlorine dioxide or peracetic acid bleaching, as even a small amount (<<5 ppm) markedly reduced pulp quality and increased reagent consumption. The results also show that evaporation can be used as a method for purifying acidic effluent from both chlorine dioxide and peracetic acid bleaching, and that the resulting condensates were free of transition metals and clean enough for reuse in both bleaching processes. The purity of the condensates was further improved by neutralising the feed pH of the acidic effluents from the bleach plant before evaporation. The results also indicate that the chloride ions in the concentrate arising from evaporation of the acidic effluent from chlorine dioxide bleaching can be removed by a combined acidification/re-evaporation system.

The popularization and health benefits associated with the &ldquo
Mediterranean diet&rdquo
saw a world wide increase in the production and consumption of processed olives and olive oil. During the brining of table olives large quantities of processed olive waste water is seasonally generated. This blackish-brown, malodours liquid is rich in organic and phenolic compounds, which cause environmental problems upon discarding. Currently, processed wastewater is discarded into large evaporation ponds where it poses serious environmental risks. The biodegradation of organic substrates present in the olive wastewater is inhibited by the high concentrations of phenolic compounds. In order to identify organisms which could potentially be used in the bioremediation of olive wastewater, 36 microbial strains were isolated from evaporation ponds in the Boland region of South Africa. Twenty five isolates were capable of growth on 50% olive wastewater and their bioremediation potential as well as their ability to produce valuable intermediate compounds were subsequently characterized. Based on the RPHPLC results, which showed that a number of chemical intermediates were produced in fermentation of olive wastewater, isolate AS-35 was selected for further analysis. Strain AS-35, identified as a Bacillus megaterium, was significantly influenced by the exposure to olive waste. The total cellular protein profile, generation time and cellular morphology of this isolate were dramatically affected by the introduction of olive waste. This study investigated the differential gene display of Bacillus megaterium following exposure to olive wastewater. Proteomic and transcriptomic differences of the organism cultured in nutrient rich LB and olive wastewater were compared. These results indicated that AS-35 expressed genes involved in glycolysis, tryptophan and nucleotide synthesis as well as the chaperones GroEL and DnaK during its growth in LB. In contrast, genes induced following the abolishment of glucose dependent catabolite repression, genes involved in biotin synthesis and ß
-oxidation of fatty or organic acids as well as a gene whose expression is regulated by stress induced sB-dependent regulon were expressed during olive waste growth.

Thesis (DTech (Chemical Engineering))--Cape Peninsula University of Technology, 2017.
Industrial wastewater management pertaining to the mining industry has become increasingly stringent, with companies being required to develop environmentally benign wastewater management practices worldwide. The industries that utilise cyanide compounds for the recovery of precious and base metals in a process known as the cyanidation process, have contributed substantially to environmental deterioration and potable water reserve contamination due to the discharge of poorly treated, or untreated, cyanide containing wastewater. Hence, a biotechnological approach was undertaken in this study to remediate free cyanide (CN-) and thiocyanate (SCN-), which are the major chemical contaminants which are normally found in cyanidation wastewaters. Furthermore, this biotechnological approach was investigated to understand the fundamental aspects of using this approach such that the information gathered can be utilized in pilot plant studies. Therefore, bioprospecting of potential CN- and SCN--degrading organisms was undertaken using two approaches; (i) culture-dependent approach and (ii) culture-independent approach. Using the culture-dependent approach, Pseudomonas aeruginosa STK 03, Exiguobacterium acetylicum and Bacillus marisflavi were isolated from an oil spill site and river sediment samples, respectively. STK 03 was evaluated for the biodegradation of CN- and SCN- under alkaline conditions. The organism had a CN- degradation efficiency of 80% and 32% from an initial concentration of 250 and 450 mg CN-/L, respectively. Additionally, the organism was able to degrade SCN-, achieving a degradation efficiency of 78% and 98% from non- and CN- spiked cultures, respectively. Furthermore, the organism was capable of heterotrophic nitrification but was unable to denitrify aerobically, with the autotrophic degradation of CN- by STK 03 being abortive.

Escherichia coli (E. coli) is a common inhabitant of surface waters in the developed and developing worlds. The majority of E. coli cells present in water are not particularly pathogenic to humans; however, there are some present in small proportion that possess virulence genes that allow them to colonize the digestive tract. Pathogenic E. coli causes acute and chronic diarrheal diseases, especially among children in developing countries and in travelers in these locales. The present study, conducted between August 2007 and July 2008, investigated the prevalence and distribution of virulent E. coli strains as either free or attached cells in the final effluents of three wastewater treatment plants located in the Eastern Cape Province of South Africa and its impact on the physico-chemical quality of the receiving water body. The wastewater treatment plants are located in urban (East Bank Reclamation Works, East London), peri-urban (Dimbaza Sewage Treatment Works) and in rural area (Alice Sewage Treatment Works). The effluent quality of the treatment plants were acceptable with respect to pH (6.9-7.8), temperature (13.8-22.0 °C), dissolved oxygen (DO) (4.9-7.8 mg/L), salinity (0.12-0.17 psu), total dissolved solids (TDS) (119-162 mg/ L) and nitrite concentration (0.1-0.4 mg/l). The other xii physicochemical parameters that did not comply with regulated standards include the following: phosphate (0.1-4.0 mg/L); chemical oxygen demand (COD) (5-211 mg/L); electrical conductivity (EC) (237-325 μS/cm) and Turbidity (7.7-62.7 NTU). Results suggest that eutrophication is intensified in the vicinity of the effluent discharge points, where phosphate and nitrate were found in high concentrations. Presumptive E. coli was isolated from the effluent samples by culture-based methods and confirmed using Polymerase Chain Reaction (PCR) techniques. Antibiogram assay was also carried out using standard in vitro methods on Mueller Hinton agar. The viable counts of presumptive E. coli for the effluent samples associated with 180 μm plankton size ranged between 0 – 4.30 × 101 cfu/ml in Dimbaza, 0 – 3.88 × 101 cfu/ml in Alice and 0 – 8.00 × 101 cfu/ml in East London. In the 60 μm plankton size category E. coli densities ranged between 0 and 4.2 × 101 cfu/ml in Dimbaza, 0 and 2.13 × 101 cfu/ml in Alice and 0 and 8.75 × 101 cfu/ml in East London. Whereas in the 20 μm plankton size category presumptive E. coli density varied from 0 to 5.0 × 101 cfu/ml in Dimbaza, 0 to 3.75 × 101 cfu/ml in Alice and 0 to 9.0 × 101 cfu/ml in East London. The free-living presumptive E. coli density ranged between 0 and 3.13 × 101 cfu/ml in Dimbaza, between 0 and 8.0 × 101 cfu/ml in Alice and between 0 and 9.5 × 101 cfu/ml in East London. Molecular analysis successfully amplified target genes (fliCH7, rfbEO157, ial and aap) which are characteristic of pathogenic E. coli strains. The PCR assays using uidA-specific primer confirmed that a genetic region homologous in size to the E. coli uidA structural gene, including the regulatory region, was present in 3 of the E. coli isolates from Alice, 10 from Dimbaza and 8 from East London. Of the 3 E. coli isolates from Alice, 1 (33.3%) was positive for the fliCH7 genes and 3 was positive for rfbEO157 genes. Out of the 10 isolates from Dimbaza, 4 were xiii positive for fliCH7 genes, 6 were positive for the rfbEO157 genes and 1 was positive for the aap genes; and of the 8 isolates from East London, 1 was positive for fliCH7 genes, 2 were for the rfbEO157 genes, 6 were positive for the ial genes. Antimicrobial susceptibility profile revealed that all of the E. coli strains isolated from the effluent water samples were resistant (R) to linezolid, polymyxin B, penicillin G and sulfamethoxazole. The E. coli isolates from Dimbaza (9/10) and East London (8/8) respectively were resistant to erythromycin. All the isolates were found to be susceptible (S) to amikacin, ceftazidime, ciprofloxacin, colistin sulphate, ceftriaxone, cefotaxime, cefuroxime, ertapenem, gatifloxacin, gentamycin, imidazole, kanamycin, meropenem, moxifloxacin, neomycin, netilmicin, norfloxacin and tobramycin. The findings of this study revealed that the Alice wastewater treatment plant was the most efficient as it produced the final effluent with the least pathogenic E. coli followed by the Dimbaza wastewater treatment plant. In addition, the findings showed that the wastewater treatment plant effluents are a veritable source of pathogenic E. coli in the Eastern Cape Province watershed. We suggest that to maximize public health protection, treated wastewater effluent quality should be diligently monitored pursuant to ensuring high quality of final effluents.

Thesis (MSc Food Sc)--Stellenbosch University, 2015.
ENGLISH ABSTRACT: The large volumes of effluent water generated by distillery industries is an issue of great concern as it contains pollutants that must be treated according to environmental legislation. It has been reported that grain distillery wastewater (GDWW) is high in fats, oils and greases (FOG) that can be reduced by treating with suitable microorganisms. The objective of this study was to investigate the biodegradability of FOG in GDWW. This was done by isolating lipolytic bacteria from soil, which was situated close to the GDWW treatment plant at a distillery in Wellington, South Africa. These isolates were screened for lipolytic activity on various fat substrates. Secondly, the most desirable isolates were subjected to batch biodegradation trials using GDWW as substrate and tested for their ability to biodegrade FOG. Each of the four isolates, Pseudomonas fluorescens (1), Pseudomonas luteola (2), Stenotrophomonas maltophilia (3) and Bacillus licheniformis (4) were screened on three types of media: DifcoTM Spirit Blue Agar with Tributyrin (SBA-Tri); Victoria Blue B Agar with Cotton Seed Oil (VBB-CSO); and Victoria Blue B Agar with GDWW (VBB-GDWW) at different temperatures (25°C, 30°C, 37°C and 50°C) to determine optimal enzyme activity for lipolysis. Lipolysis was taken as positive when growth of dark blue colonies was formed or by the formation of a clear zone around the colony. Lipolysis was observed at all the aforementioned temperatures for P. fluorescens, P. luteola and S. maltophilia. Bacillus licheniformis failed to show any lipolytic activity at 50°C on the SBA-Tri. A decrease in lipolytic (clear) zone was observed at an increase in temperature from 25°C to 37°C for P. fluorescens. When VBB-GDWW was used as lipid substrate, isolates failed to indicate any clear zone of lipolysis, however, growth was present for all isolates in the form of a dark blue zone around colonies, which were also positive for lipolytic activity. Three lipolytic bacteria (P. luteola, S. maltophilia, and B. licheniformis) isolated from the above study were subjected to GDWW of various FOG concentrations (70 – 211 mg.L-1). These isolates were allowed to acclimatise to GDWW during a batch biodegradation period (18 – 21 d) at 37°C. Bacillus licheniformis showed the highest FOG reduction of 83% after 18 d exposure. All the strains showed that an initial acclimatisation phase improved the biodegradation of the FOG. A fatty acid profile was obtained for each batch biodegradation trial after the acclimatisation phase. It was found that these strains either biodegraded the fatty acids (FAs) or, as in the case of P. luteola, formed myristic and pentadecyclic acids from free FAs. The formation of FAs may have occurred through a process of inter-esterification. It was also found that certain precursors such as palmitoleic acid might be formed under aerobic or anaerobic conditions. In this study it was shown that biodegradation of FOG can be improved by an initial acclimatisation period. Single cultures with the desirable properties can be used to lower the FOG in GDWW and need not be used in mixed cultures that could produce inhibitory components that would otherwise upset the biodegradation activity of isolates present. Bacillus licheniformis could be used as a FOG-degrading isolate during the treatment of wastewaters high in FOG. However, future studies should focus on bioaugmenting the FOG degrading bacteria from this study with other strains to monitor its activity and ensure survival and activity in larger scale studies.
AFRIKAANSE OPSOMMING: Die groot volumes afloopwater wat opgelewer word deur die distilleer-industrie is ‘n kwessie wat groot kommer wek aangesien dit groot hoeveelhede besoedelende stowwe bevat. Daarom moet dit, volgens omgewingsverwante wetgewing, behandel word. Daar is voorheen gerapporteer dat graandistillerings-afloopwater (GDAW) hoog is in vette, olies en ghries (VOG) en dat hierdie VOG verminder kan word deur die GDAW te behandel met toepaslike mikroörganismes. Die oorhoofse doelstelling van hierdie studie was om die bioafbreekbaarheid van die VOG in GDAW te ondersoek. Dit is eerstens gedoen deur lipolitiese bakterieë uit grond wat naby ‘n graandistillerings-aanleg (Wellington, SuidAfrika) geleë is, te isoleer. Verskeie vetsubstrate is gebruik om hierdie isolate vir lipolitiese aktiwiteit te toets. Tweedens is die verkose isolate getoets vir lipolitiese aktiwiteit deur gebruik te maak van lot-bio-afbreekbaarheidsmetode. Tydens hierdie metode is GDAW as substraat gebruik en die verskillende bakterieë se vermoë om VOG af te breek is getoets. Om die optimale ensiemaktiwiteit vir lipolise van elk van die vier isolate nl. Pseudomonas fluorescens (1), Pseudomonas luteola (2), Stenotrophomonas maltophilia (3) en Bacillus licheniformis (4), vas te stel, is elk getoets op drie verkillende media: “DifcoTM Spirit Blue Agar” met Tributirien (SBA-Tri); “Victoria Blue B Agar” met Katoensaadolie (VBB-KSO); en “Victoria Blue B Agar” met GDAW (VBB-GDAW) teen verskillende temperature (25°C, 30°C, 37°C en 50°C). Indien donker-blou kolonies gevorm is of ‘n deursigbare sone rondom ‘n kolonie waargeneem is, is lipolise as “positief” beskou. Lipolise is waargeneem teen alle voorafgenoemde temperature vir P. fluorescens, P. luteola en S. maltophilia. Bacillus licheniformis het nie lipolitiese aktiwiteit getoon teen 50°C op SBA-Tri. ‘n Afname in die deursigbare sone is waargeneem teenoor ‘n toename in temperatuur vanaf 25°C tot 37°C vir P. fluorescens. In die geval van VBB-GDAW as lipiedsubstraat, het isolate geen deursigbare sone vir lipolise getoon nie. Daar was egter ‘n donker-blou sone rondom kolonies teenwoordig, wat ook positief is vir lipolitiese aktiwiteit. Drie lipolitiese bakterieë (P. luteola, S. maltophilia, and B. licheniformis) is geïsoleer uit bogenoemde studie en is aan inkubasie in GDAW teen verksillende VOGkonsentrasies (70 – 211 mg.L-1) blootgestel. Hierdie isolate is toegelaat om te akklimatiseer tot die GDAW tydens ‘n lot-bio-afbreekbaarheidstydperk (18 – 21 d) teen 37°C. Bacillus licheniformis het die hoogste VOG-afname van 83% na 18 d blootstelling getoon. Alle bakterieë het getoon dat ‘n aanvanklike akklimatiserings-tydperk die bioafbreekbaarheid van die VOG verbeter. ‘n Vetsuur-profiel is verkry vir elk van die lot-bio afbreekbaarheidstoetse na die akklimatiserings-fase. Daar is bevind dat hierdie bakterieë óf die vetsure afgebreek het óf, soos in die geval van P. luteola, miristiese en pentadesikliese sure, vanaf vry-vetsure, gevorm het. Die vorming van vetsure is moontlik as gevolg van die proses van inter-esterifikasie. Dit is verder bevind dat sekere voorlopers, soos palmitoë-oleïensuur, gevorm kan word onder aërobies of anaërobiese toestande. In hierdie studie is getoon dan die bio-afbreekbaarheid van VOG verbeter kan word deur ‘n aanvanklike akklimatiserings-tydperk toe te pas. Enkel-kulture met die verkose eienskappe kan gebruik word om die VOG in GDAW te verminder. Gemengde kulture, wat inhiberende komponente produseer wat moontlik die bio-afbreekbaarheids proses negatief kan beïnvloed, hoef dus nie gebruik te word nie. Bacillus licheniformis kan gebruik word as ‘n VOG-afbrekende isolaat tydens die behandeling van afloopwater wat hoog in VOG is. Verdere studies moet egter fokus op die samevoeging van VOGafbrekende bakterieë vanuit hierdie studie asook ander bakterieë om die aktiwiteit daarvan te monitor en sodoende oorlewing en aktiwteit op ‘n groter skaal te verseker.

Thesis submitted in fulfilment of the requirements for the degree Master of Technology: Chemical Engineering in the Faculty of Engineering at the Cape Peninsula University of Technology 2014
Environmental legislation focusing on wastewater disposal in industries that utilise cyanide and/or cyanide-related compounds has become increasingly stringent worldwide, with many companies that utilise cyanide products required to abide by the Cyanide International Code associated with the approval of process certifications and management of industries which utilise cyanide. This code enforces the treatment or recycling of cyanide-contaminated wastewater. Industries such as those involved in mineral processing, photo finishing, metal plating, coal processing, synthetic fibre production, and extraction of precious metals, that is, gold and silver, contribute significantly to cyanide contamination in the environment through wastewater. As fresh water reserves throughout the world are low, cyanide contamination in water reserves threatens not only the economy, but also endangers the lives of living organisms that feed from these sources, including humans. In the mining industry, dilute cyanide solutions are utilised for the recovery of base (e.g. Cu, Zn, Ni, etc.) and precious metals (e.g. Au, Ag, etc.). However, for technical reasons, the water utilised for these processes cannot be recycled upstream of the mineral bioleaching circuit as the microorganisms employed in mineral bioleaching are sensitive to cyanide and its complexes, and thus the presence of such compounds would inhibit microbial activity, resulting in poor mineral oxidation. The inability to recycle the water has negative implications for water conservation and re-use, especially in arid regions. A number of treatment methods have been developed to remediate cyanide containing wastewaters. However, these chemical and physical methods are capital intensive and produce excess sludge which requires additional treatment. Furthermore, the by-products that are produced through these methods are hazardous. Therefore, there is a need for the development of alternative methods that are robust and economically viable for the bioremediation of cyanide-contaminated wastewater. Biological treatment of free cyanide in industrial wastewaters has been proved a viable and robust method for treatment of wastewaters containing cyanide. Several bacterial species, including Bacillus sp., can degrade cyanide to less toxic products, as these microorganisms are able to use the cyanide as a nitrogen source, producing ammonia and carbon dioxide. These bacterial species secrete enzymes that catalyse the degradation of cyanide into several end-products. The end-products of biodegradation can then be utilised by the microorganisms as nutrient sources. This study focused on the isolation and identification of bacterial species in wastewater containing elevated concentrations of cyanide, and the assessment of the cyanide biodegradation ability of the isolates. Thirteen bacterial isolates were isolated from electroplating wastewater by suppressing the growth of fungal organisms and these species were identified as species belonging to the Bacillus genus using the 16S rDNA gene. A mixed culture of the isolates was cultured in nutrient broth for 48 hours at 37°C, to which FCN as KCN was added to evaluate the species‟ ability to tolerate and biodegrade cyanide in batch bioreactors. Subsequently, cultures were supplemented solely with agro-waste extracts, that is, Ananas comosus extract (1% v/v), Beta vulgaris extract (1% v/v), Ipomea batatas extract (1% v/v), spent brewer‟s yeast (1% v/v) and whey (0.5% w/v), as the primary carbon sources. Owing to the formation of high ammonium concentration from the cyanide biodegradation process, the nitrification and aerobic denitrification ability of the isolates, classified as cyanide-degrading bacteria (CDB) was evaluated in a batch and pneumatic bioreactor in comparison with ammonia-oxidising bacteria (AOB). Furthermore, the effects of F-CN on the nitrification and aerobic denitrification was evaluated assess the impact of F-CN presence on nitrification. Additionally, optimisation of culture conditions with reference to temperature, pH and substrate concentration was evaluated using response surface methodology. Using the optimised data, a continuous biodegradation process was carried out in a dual-stage packed- bed reactor combined with a pneumatic bioreactor for the biodegradation of F-CN and subsequent nitrification and aerobic denitrification of the formed ammonium and nitrates. The isolated bacterial species were found to be gram positive and were able to produce endospores that were centrally located; using the 16S rDNA gene, the species were found to belong to the Bacillus genus. The species were able to degrade high cyanide concentration in nutrient broth with degradation efficiencies of 87.6%, 65.4%, 57.0% and 43.6% from 100 mg F-CN/L, 200 mg F-CN/L, 300 mg F-CN/L, 400 mg F-CN/L and 500 mg F-CN/L respectively over a period of 8 days. Additionally, the isolates were able to degrade cyanide in an agro-waste supported medium, especially in a medium that was supplemented with whey which achieved a degradation efficiency of 90% and 60% from 200 mg F-CN/L and 400 mg F-CN/L, respectively over a period of 5 days. The nitrification ability of the isolates was evaluated and the removal of NH4 +/NO3 - by the CDB and AOB in both shake flasks and pneumatic bioreactor was determined to be pH dependent. The maximum NH4 +/NO3 - removal evaluated over a period of 8 days for CDB and 15 days for AOB, observed at pH 7.7 in shake flasks, was 75% and 88%, respectively, in the absence of F-CN. Similarly, the removal of NH4 +/NO3 - in a pneumatic bioreactor was found to be 97.31% for CDB and 92% for AOB, thus demonstrating the importance of aeration in the designed process. The nitrification by CDB was not inhibited by cyanide loading up to a concentration of 8 mg FCN/ L, while the AOB were inhibited at cyanide loading concentration of 1 mg F-CN/L. The CDB removed the NH4 +/NO3 - in PBSs operated in a fed-batch mode, obtaining efficiencies >99% (NH4 +) and 76 to 98% (NO3 -) in repeated cycles (n = 3) under F-CN (≤8 mg F-CN/L). The input variables, that is, pH, temperature and whey-waste concentration, were optimised using a numerical optimisation technique where the optimum conditions were found to be: pH 9.88, temperature 33.60 °C and whey-waste concentration 14.27 g/L, under which 206.53 mg CN-/L in 96 h can be biodegraded by the microbial species from an initial cyanide concentration of 500 mg F-CN/L. Furthermore, using the optimised data, cyanide biodegradation in a continuous mode was evaluated in a dual-stage packed-bed bioreactor connected in series to a pneumatic bioreactor system used for simultaneous nitrification including aerobic denitrification. The whey-supported Bacillus sp. culture was not inhibited by the free cyanide concentration of up to 500 mg F-CN/L, with an overall degradation efficiency of ≥99% with subsequent nitrification and aerobic denitrification of the formed ammoniu and nitrates over a period of 80 days.

The production of final effluents that meet discharged requirements and guidelines remain a major challenge particularly in the developing world with the resultant problem of surface water pollution. This study assessed the physicochemical and microbiological qualities of two wastewater treatment works in the Eastern Cape Province of South Africa in terms of the prevalence of faecal coliforms and Escherichia coli O157:H7 over a five month period. All physicochemical and microbiological analyses were carried out using standard methods. Data were collected in triplicates and analysed statistically using IBM SPSS version 20.0. The ranges of some of the physicochemical parameters that complied with set guidelines include pH (6.7 – 7.6), TDS (107 – 171 mg/L), EC (168 – 266 μS/cm), Temperature (15 – 24oC), NO3- (0 – 8.2 mg/L), NO2- (0.14 – 0.71 mg/L) and PO4 (1.05 – 4.50 mg/L). Others including Turbidity (2.64 – 58.00 NTU), Free Cl (0.13 – 0.65 mg/L), DO (2.20 – 8.48 mg/L), BOD (0.13 – 6.85 mg/L) and COD (40 – 482 mg/L) did not comply with set guidelines. The microbiological parameters ranged 0 – 2.7 × 104 CFU/100 ml for FC and 0 – 9.3 × 103 for EHEC CFU/100 ml, an indication of non-compliance with set guidelines. Preliminary identification of 40 randomly selected presumptive enterohemorrhagic E. coli isolates by Gram’s staining and oxidase test shows 100% (all 40 selected isolates) to be Gram positive while 90% (36 randomly selected isolates) were oxidase negative. Statistical correlation between the physicochemical and the microbiological parameters were generally weak except in the case of free chlorine and DO where they showed inverse correlation with the microbiological parameters. The recovery of EHEC showed the inefficiency of the treatment processes to effectively inactivate the bacteria, and possibly other pathogenic bacteria that may be present in the treated wastewater. The assessment suggested the need for proper monitoring and a review of the treatment procedures used at these treatment works.

Wastewater discharges may contain health compromising pathogens and carcinogenic and/or chemical substances that could compromise the public health and impact negatively on the environment. The present study was conducted between August 2007 and July 2008 to evaluate the Listeria abundance (as free-living and plankton associated species) and physicochemical qualities of the final effluents of three wastewater treatment facilities in the Eastern Cape Province of South Africa selected to represent typical urban, peri-urban and rural communities and the impact of the discharged final effluents on their respective receiving watershed, as well as to elucidated the in vitro antibiotic susceptibilities and resistance genes profile of Listeria species isolated from the final effluents. The suitability of the secondary effluent of the urban treatment facility (as a case study) for use in agriculture and aquaculture with reference to recommended standards was also determined. Wastewater samples were collected from the raw sewage, secondary effluent, final treated effluent, discharge point, 500 m upstream discharge point, and 500 m downstream discharge point from all three locations on a monthly basis throughout the study period. Listeria abundance in the final effluents and the receiving watersheds varied between 2.9× 100 and 3.52 × 105cfu/ml across the sampled locations. Free-living listerial density across the sampled locations ranged between 0 and 3.2 × 103cfu/ml while counts of Listeria species attached to large (180 μm) planktons varied from 0 to 1.58 × 105 cfu/ml and those of the 60 and 20 μm categories were in the range of 0 to 1.32 × 103 cfu/ml and 0 to 2.82 × 105 cfu/ml respectively. Listeria abundance did not vary significantly with location and season; there was however, significant (P < 0.05; P < 0.01) variance in Listeria abundance with plankton sizes across the locations. Free-living Listeria species were more abundant in the rural and urban xii communities than plankton attached Listeria species; whereas the reverse was the case in the peri-urban community. Prevalence of Listeria in terms of total counts was 100 percent across all sampled locations. Free-living Listeria species showed prevalence ranging from 84-96 percent across the sampling locations; while Listeria species attached to large (180 μm) planktons exhibited prevalence ranging from 75 percent to 90 percent. The prevalence of medium-sized (60 μm) plankton associated Listeria species varied between 58 percent and 92.5 percent; whereas those of Listeria species attached to small (20 μm) planktons ranged from 65-100 percent across all three communities. Listeria prevalence was generally a reflection of the turbidity of the water system, with free-living Listeria species being more prevalent than plankton associated cells in the relatively less turbid rural and urban waters compared to the more turbid peri-urban waters where plankton attached cells were more prevalent in comparison with their free living counterparts The final treated effluent quality fell short of recommended standards for turbidity, chemical oxygen demand and phosphate across all three communities. In addition, the final effluent of the rural treatment plant also fell short of recommended standard for NO3, while that of the urban treatment plant did not comply with acceptable limits for dissolved oxygen and nitrite. Other physicochemical parameters were compliant with set standards after treatment. An inverse relationship was observed between chlorine residual and listerial density across the sampled facilities; the effect of chlorine was however not enough to eliminate the pathogen from the water systems. At the urban treatment plant and its receiving watershed, pH, temperature, EC, turbidity, TDS, DO, and nitrate varied significantly with season and sampling point (P < 0.05; P < 0.01). Salinity also varied significantly with sampling point (P < 0.01), while COD and nitrite varied significantly with season (P < 0.05). Although, the treated effluent fell within recommended water quality standard for pH, TDS, nitrate and nitrite, it fell short of stipulated standards for other parameters. Whereas the microbial quality of the secondary treated effluent at this (urban) facility fell short of recommended standard after secondary treatment, its physicochemical quality were generally compliant with recommended standards for reuse wastewater in agriculture and aquaculture. Listeria pathogens isolated from effluents of the rural wastewater facility were sensitive to 11 (55 percent) of the 20 test antibiotics, and showed varying (7-71 percent) levels of resistance to 8 antibiotics; whereas those isolated from the peri-urban community showed sensitivity to 6 (30 percent) of the 20 test antibiotics, and varying (6-94 percent) levels of resistance to 12 antibiotics; while the urban effluent isolates were sensitive to 3 (15 percent) of the 20 test antibiotics, and showed varying (4.5-91 percent) levels of resistance to 17 antibiotics. Multiple antibiotic resistances involving 78.5-100 percent of isolates and antibiotics combination ranging from 2-10 antibiotics was observed across the sampled locations. Penicillin G and ampicillin showed remarkably high (64-91 percent) phenotypic resistance across the three sampled facilities. Other antibiotics, to which isolates showed significant resistance, were linezolid (22-88 percent); erythromycin (43-94 percent) and sulphamethoxazole (7-94 percent). Two of the 14 Listeria strains isolated from the rural effluents were positive for ereA and sul1 antibiotic resistance genes; while sulII genes were detected in five of the 23 Listeria isolates from the urban effluent and none was detected in isolates from the peri-urban community. The presence of antimicrobial resistance genes in the isolates did not correlate with phenotypic antibiotic resistance. The current study demonstrated that Listeria pathogens easily survived the activated sludge treatment process as free-living and plankton attached entities and suggests that municipal wastewater treatment plants are a significant source of multiple resistant Listeria pathogens in the South African aquatic milieu. While the physicochemical quality of the urban final effluent suggests that it is a major source of pollution to the receiving watershed, the secondary effluent quality demonstrated a great potential for use in agriculture and aquaculture.

Thesis (Msc Food Sc (Food Science))--University of Stellenbosch, 2007.
The distillery industry generates large volumes of heavily polluted wastewater and thus effective wastewater treatment is essential. It has been reported that a chemical oxygen demand (COD) reduction of more than 90% can be achieved when wine distillery wastewater (WDWW) is treated in an upflow anaerobic sludge blanket (UASB) reactor. The first objective of this study was to investigate UASB treatment of WDWW and to try to enhance the efficiency by using ozonation treatments. Secondly, the impact of grain distillery wastewater (GDWW) on UASB granules was determined. The third objective was to determine whether ozonation and enzymatic treatment combinations might improve the biodegradability of GDWW and thus make GDWW more amenable to UASB treatment. It was found that UASB treatment combined with ozonation improved the WDWW treatment efficiency. When diluted WDWW (chemical oxygen demand COD = 4 000 mg.L-1) was ozonated (dose = 47 mg.L-1) in a 50 L venturi circulating contactor system, the COD reduction was 7%. When WDWW was treated in a laboratory-scale UASB reactor (substrate pH = 7.0, COD = 4 000 mg.L-1 and organic loading rate (OLR) = 4.0 kg COD.m-3.d-1), the COD reduction was 92%. When the UASB treatment was combined with either pre- or postozonation, the COD reduction was 94 and 96%, respectively. When UASB treatment was combined with pre- and post-ozonation, a COD reduction of 98% was achieved. The activity of the UASB granules was also found to improve over time, despite the addition of the ozonation treatment. It has been reported that operational problems occur when GDWW is treated in an UASB reactor as a result of the encapsulation of the granules. This was confirmed when granules from a full-scale UASB treating WDWW became encapsulated in a layer after being exposed to GDWW (COD = 4 000 mg.L-1) for 24 d. The results showed that the lipid content of the granules increased from 1.25 to 60.35 mg lipid.g-1 granule over the 24 d exposure period. Therefore, granules exposed to GDWW were encapsulated in a lipid-rich layer and as a result the contact between the GDWW and microbial consortium in the granules was reduced. The operational problems found during the industrial UASB treatment of GDWW were ascribed to the encapsulation of the granules. Combinations of ozonation (dose = 1 476 mg.L-1) generated in a 2 L bubble column and enzymatic treatments (1% FogFreeTM (FF) dosage and 2 d incubation at 35°C) were found to improve the biodegradability of GDWW. This improvement was in terms of lipid reduction in GDWW, granule activity and visual appearance of the encapsulating layer of the granules. The highest lipid reduction (90%), highest granule activity, lowest lipid content of the granules (3.74 ± 0.10 mg.g-1 granule) and best visual appearance were achieved in ozonated GDWW treated with 1% FF, followed by just ozonation. The higher lipid reduction and subsequent higher granule activity were ascribed to the reduction in lipids which resulted in the fact that fewer lipids were available to encapsulate the granules. As a result of the lipid reduction, the granule activity improved and the GDWW was made more amenable to UASB treatment. This study proved that UASB treatment combined with ozonation led to an enhancement of the treatment efficiency of WDWW. It was also found that the cause of the operational problems during UASB treatment of GDWW was as a result of the granules being encapsulated in a lipid-rich layer. It was established that treating GDWW prior to UASB treatment improved the biodegradability of GDWW. The data from the study showed that high lipid reduction in the GDWW directly led to better granule activity, lower granule lipid content and a thinner encapsulating layer. Based on the data from this study, it is recommended that GDWW be ozonated prior to other treatments because it can be done inline and the costs would be lower than that of enzymatic treatments.

South Africa is currently experiencing water shortage crisis, a challenge that has been attributed not only to the scarcity of freshwater, but also to fast degrading water quality. Factors such as rapid urbanisation, population and economic growth, climate change as well as poor operational and maintenance of many of the exisiting water/wastewater treatment facilities have been acknowledged as important contributors to degrading water quality in the country. Untreated or inadequately treated discharged wastewater effluents constitute point source pollution to many freshwater environments in South Africa. Hence, it becomes imperative to evaluate wastewater discharges in other to protect the scarce freshwater resource, the environment and public health. Over a twelve-month sampling period (September 2012 to August 2013), we assessed the bacteriological, virological and physicochemical qualities of the discharged final effluents of two wastewater treatment facilities in the Eastern Cape Province of South Africa. For the physicochemical assessment, a total of 144 final effluent samples were collected from both the final effluent tanks (FE) and the discharge points (DP) of the treatment facilities. Physicochemical parameters including pH, temperature, turbidity, total dissolved solids (TDS), dissolved oxygen (DO), electrical conductivity (EC) and free chlorine concentration were determined on site while biological oxygen demand (BOD), nitrate (NO3-), nitrite (NO2-), phosphate (PO4-) and chemical oxygen demand (COD) were determined in the laboratory. The bacteriological analysis of the samples was done using standard membrane filtration (MF) technique. Bacterial group assessed included: faecal indicator bacteria (faecal coliforms and E. coli) and Vibrio species, while the antibiotic susceptibility profiles of selected E. coli and Vibrio species isolates against some selected antibiotics commonly used in human therapy and veterinary medicine were determind using the standard agar-disc diffusion method. The occurrence and concentrations of human enteric viruses including: human adenovirus (HAdV), hepatitis A virus (HAV) and rotavirus (RoV) in the samples were determined by TaqMan-based real-time polymerase chain reaction (qPCR) following concentration by adsorption-elution method. The physicochemical characteristics of the samples ranged as follows: pH (6.5 – 7.6), TDS (95 – 171 mg/L), EC (134 – 267 μS/cm), temperature (12 – 27 °C), turbidity (1.5 – 65.7 mg/L), free chlorine (0.08 – 0.72 mg/L), DO (2.06 – 9.81 mg/L), BOD (0.13 – 9.81 mg/L), NO3- (0 – 21.5 mg/L), NO2- (0 – 0.72 mg/L), PO4- (0 – 18.3 mg/L) and COD (27 – 680 mg/L). Some of the characteristic such as pH, TDS, EC, temperature, nitrite and DO (on most instances) complied with recommended guidelines. Other characteristics, however, including turbidity, BOD, nitrate, phosphate and COD fell short of the recommended guidelines. All the 48 samples analysed for bacteriological qualities tested positive for the presence of the bacterial groups with significant (P≤0.05) seasonal variation in their densities. Faecal coliforms were detected in counts ranging from 1 CFU/100ml to 2.7 × 104 CFU/100ml. Presumptive E. coli counts ranged generally between 1 CFU/100ml – 1.4 × 105 CFU/100ml while counts of presumptive Vibrio species ranged between 4 CFU/100ml – 1.4 × 104 CFU/100ml. Molecular identification of the presumptive isolates by polymerase chain reactions PCR gave positive reaction rates of 76.2 percent (381/500) and 69.8 percent (279/400) for E.coli and Vibrio species respectively. The antibiotic susceptibility profiling of 205 PCR-confirmed Vibiro isolates against 18 commomly used antibiotics showed resistance frequencies ranging from 0.5 percent (imipenem) to 96.1 percent (penicillin G) at recommended breakpoint concentrations. Eighty-one percent (166/205) of the Vibrio isolates showed multidrug resistance (resistance to 3 or more antibiotics) with the most common multiple antibiotic resistance phenotype (MARP) being AP-T-TM-SMX-PG-NI-PB, occurring in 8 isolates.

Worldwide, water reuse is promoted as an alternative for water scarcity, however, wastewater effluents have been reported as possible contaminants to surface water. The failure of some wastewater treatment processes to completely remove organic matter and some pathogenic microorganisms allows them to initiate infections. This manifests more in communities where surface water is used directly for drinking. To assess water quality, bacteria alone cannot be used as it may be absent in virus-contaminated water. This study was carried out to assess the quality of two wastewater treatment plant effluents from the Eastern Cape Province of South Africa. Physicochemical parameters and microbiological parameters like faecal coliforms, adenovirus, rotavirus, hepatitis A virus, norovirus and enterovirus were evaluated over a projected period of one year. Physicochemical parameters were measured on site using multiparameters, faecal coliforms enumerated using culture-based methods and viruses are detected using both conventional and real-time PCR. Physicochemical parameters like electrical conductivity, turbidity, free chlorine and phosphates were incompliant with the standards set by the Department of Water affairs for effluents to be discharged. Faecal coliform counts were nil for one plant (WWTP-R) where they correlated inversely (P < 0.01) with the high free chlorine. For WWTP-K, faecal coliforms were detected in 27% of samples in the range of 9.9 × 101 to 6.4× 104 CFU/100ml. From the five viruses assessed, three viruses were detected with Rotavirus being the most abundant (0-2034176 genome copies/L) followed by Adenovirus (0–275 genome copies/L) then Hepatitis A virus (0–71 genome copies/L) in the WWTP-K while none of the viruses was detected in WWTP-R. Species B, species C and Adv41 serotypes were detected from the May 2013 and June 2013 samples where almost all parameters were incompliant in the plant. The detection of these viruses in supposedly treated effluents is suggestive of these being the sources of contamination to surface water and therefore renders surface waters unsafe for direct use and to aquatic life. Although real-time PCR is more sensitive and reliable in detection of viruses, use of cell-culture techniques in this study would have been more efficient in confirming the infectivity of the viruses detected, hence the recommendation of these techniques in future projects of this nature.

Vibrio infections remain a serious threat to public health. In the last decade, Vibrio disease outbreaks have created a painful awareness of the personal, economic, societal, and public health costs associated with the impact of contaminated water in the aquatic milieu. This study was therefore designed to assess the prevalence of Vibrio pathogens in the final effluents of wastewater treatment plants (WWTPs) in the Eastern Cape Province, as well as their abilities to survive the treatment processes of the activated sludge system either as free cells or as plankton-associated entities in relation to the physicochemical qualities of the effluents. Three wastewater treatment facilities were selected to represent typical urban, sub-urban and rural communities, and samples were collected monthly from August 2007 to July 2008 from the final effluent, discharge point, 500 meter upstream and downstream of the discharge points and analysed for physicochemical parameters, Vibrio pathogens prevalence and their antibiogram characteristics using both culture based and molecular techniques. Physicochemical parameters measured include pH, temperature, electrical conductivity, salinity, turbidity, total dissolved solid (TDS), dissolved oxygen (DO), chemical oxygen demand (COD), nitrate, nitrite and orthophosphate levels. Unacceptably high levels of the assayed parameters were observed in many cases for COD (<10 - 1180 mg/l), nitrate (0.08 - 13.14 mg NO3- as N/l), nitrite (0.06 - 6.78 mg NO2- as N/l), orthophosphate (0.07-4.81 mg PO43- as P/l), DO (1.24 - 11.22 mg/l) and turbidity (2.04 -159.06 NTU). Temperature, COD and nitrite varied significantly with season (P < 0.05), while pH, EC, salinity, TDS, COD, and nitrate all varied significantly with sampling site (P < 0.01; P < 0.05). In the rural wastewater treatment facility, free-living Vibrio densities varied from 0 to 3.45 × 101 cfu ml-1, while the plankton-associated Vibrio densities vary with plankton sizes as follows: 180 μm (0 – 4.50 × 103 cfu ml-1); 60 μm (0 – 4.86 × 103 cfu ml-1); 20 μm (0 – 1.9 × 105 cfu ml-1). The seasonal variations in the Vibrio densities in the 180 and 60 μm plankton size samples were significant (P < 0.05), while the 20 μm plankton size and free-living vibrios densities were not. Molecular confirmation of the presumptive vibrios isolates revealed V. fluvialis (36.5 percent), as the predominant species, followed by V. vulnificus (34.6 percent), and V. parahaemolyticus (23.1 percent), and V. metschnikovii (5.8 percent) (detected using only API 20 NE), suggesting high incidence of pathogenic Vibrio species in the final effluent of the wastewater facility. Correlation analysis suggested that the concentration of Vibrio species correlated negatively with salinity and temperature (P < 0.001 and P < 0.002 respectively) as well as with pH and turbidity (P < 0.001), in the final effluent. Population density of total Vibrio ranged from 2.1 × 101 to 4.36 × 104 cfu ml-1 and from 2.80 ×101 to 1.80 × 105 cfu ml-1 for the sub-urban and urban communities treatment facilities respectively. Vibrio species associated with 180 μm, 60 μm, and 20 μm plankton sizes, were observed at densities of 0 - 1.36 × 103 cfu ml-1, 0 - 8.40 × 102 cfu ml-1 and 0 - 6.80 × 102 cfu ml-1 respectively at the sub-urban community‘s WWTP. In the urban community, counts of culturable vibrios ranged from 0 - 2.80 × 102 cfu ml-1 (180 μm); 0 - 6.60 × 102 cfu ml-1 (60 μm) and 0 -1.80 × 103 cfu ml-1 (20 μm). Abundance of free-living Vibrio species varied between 0 and the orders of 102 and 103 cfu ml-1 in the sub-urban and urban communities WWTPs respectively. Molecular confirmation of the presumptive vibrios isolates revealed the presence of V. fluvialis (41.38 percent), V. vulnificus (34.48 percent), and V. parahaemolyticus (24.14 percent) in the sub-urban community effluents. In the urban community V. fluvialis (40 percent), V. vulnificus (36 percent), and V. parahaemolyticus (24 percent) were detected. There was no significant correlation between Vibrio abundance and season, either as free-living or plankton-associated entities, while Vibrio species abundance correlated positively with temperature (r = 0.565; P < 0.01), salinity and dissolved oxygen (P < 0.05). Turbidity and pH showed significant seasonal variation (P < 0.05) in both locations. The Vibrio strains showed the typical multi-antibiotic-resistance of an SXT element. They were resistant to sulfamethoxazole (Sul), trimethoprim (Tmp), cotrimoxazole (Cot), chloramphenicol (Chl) and streptomycin (Str), as well as other antibiotics such as ampicillin (Amp), penicillin (Pen), erythromycin (Ery), tetracycline (Tet), nalidixic acid (Nal), and gentamicin (Gen). The antibiotic resistance genes detected includes dfr18 and dfrA1 for trimethoprim; tetA, strB, floR, sul2 blaP1, for tetracycline, streptomycin, chloramphenicol, sulfamethoxazole and β-lactams respectively. A number of these genes were only recently described from clinical isolates, demonstrating genetic exchange between clinical and environmental Vibrio species. This study revealed that there was an adverse impact on the physicochemical characteristics of the receiving watershed as a result of the discharge of inadequately treated effluents from the wastewater treatment facilities. The occurrence of Vibrio species as plankton-associated entities confirms the role of plankton as potential reservoir for this pathogen. Also the treated final effluents are reservoirs of various antibiotics resistance genes. This could pose significant health and environmental risk to the biotic component of the environment including communities that rely on the receiving water for domestic purposes and may also affect the health status of the aquatic milieu in the receiving water. There is need for consistent monitoring programme by appropriate regulatory agencies to ensure compliance of the wastewater treatment facilities to regulatory effluent quality standards.

Water is an essential need that stimulates health and well being. Increase in population size and urbanization negatively affect water resources due to high demands of effluent outputs. Wastewater is an important reservoir for Escherichia coli and can present significant acute toxicity if released into receiving water sources without being adequately treated. E. coli is used as indicator organism for the detection of faecal contamination. These strains have been considered to be one of the primary causes of diarrhoeal infections worldwide. The present study was conducted between September 2012 and June 2013 to assess the prevalence of pathogenic E. coli strains in the final effluents of four wastewater treatment plants in Chris Hani and Buffalo City Municipalities in the Eastern Cape Province of South Africa. Standard membrane filtration technique was used for bacteriological analysis and molecular based technique was used for identification of E. coli pathotypes. The results were recorded in colony forming units/100 ml. Faecal coliforms ranged between 0-9.6×10³ CFU/100 ml for the wwtp-Q and E. coli densities ranged between 0-8.4×10³ CFU/100ml. Faecal coliforms ranged between 4×10²-9.7×10³ CFU/100 ml for wwtp-M and E. coli densities ranged between 1.2×10¹-8.4×10³ CFU/100 ml. The wwtp-E showed to have bacterial counts of faecal coliforms ranging between 4.0×10³-8.2×10³ CFU/100 ml and E. coli densities ranging between 3.5×10¹-7.1×10³ CFU/100 ml. The WWTP-K in this study was only assessed for the presence of E. coli. Faecal coliforms were assessed by the other members of the group. This plant showed to have E. coli densities ranging between 0-7.5×10²CFU/100 ml. A total of 200 presumptive E. coli isolates were subjected to screening by conventional PCR in which (29%) of the wwtp-M isolates were positively identified as E. coli, (16%) of the wwtp-K, (22%) of the wwtp-Q and (34%) of the wwtp-E isolates were positively confirmed as E. coli. A total of 100 randomly selected E. coli isolates were characterised into different pathotypes. (16%) of positive isolates were detected as EPEC and 11% were detected as UPEC strains. There was no detection for the ETEC strains. Antibiotic susceptibility patterns of E. coli strains showed high levels of resistance to Penicillin G, Erythromycin, Tetracycline and Sulfamethoxazole. High levels of Susceptibility were observed in antibiotics such as Chloramphenicol, Amoxicillin and Tetracycline. The results of this study reveal that the plants were above the recommended Standard limit of zero CFU/100 ml for effluents meant to be discharge into receiving water sources. This study reveals inadequacy of the plants studied to produce effluents of acceptable quality.

L'assainissement non collectif concerne 12 à 15 millions de personnes en France. La filière classique de ce mode d’assainissement se compose généralement d'un prétraitement anaérobie par une fosse septique recevant l’ensemble des eaux usées domestiques suivi d’un système d’infiltration dans le sol ou d’un filtre à sable. Le filtre à sable vertical drainé met à profit le pourvoir épuratoire qui est principalement lié à la présence d’une biomasse sous forme d’un biofilm. Cette dynamique de la croissance de la biomasse ou du biofilm est soumise à l’impact de la nature de matériaux filtrants. L’écoulement insaturé dans ces systèmes conditionne également cette croissance du biofilm. Dans ce contexte, l'objectif du travail de la thèse est d’appréhender les mécanismes mis en jeu et particulièrement l’impact des matériaux dans le fonctionnement des filtres en comparant notamment deux types de matériaux: les sables de rivière et les agrégats concassés. Pour cela, une étude expérimental sur une unité pilote composé des réacteurs de filtration du diamètre de 30cm et différents épaisseurs de garnissage (15, 30 et 70cm) a été construite. Les réacteurs garnis de deux sables roulés et deux agrégats concassés, sont alimentés en effluent septique avec une charge volumique 12cm/jour par 10 bâchés par jour. Suite des matériaux, une étude de la performance épuratoire avec le suivi des composants biochimiques de la biomasse totale et de la matrice extracellulaire du biofilm est réalisée en comparant notamment les deux types de matériaux filtrants
The onsite wastewater treatment systems concern 12 to 15 million of people in France. The treatment plant is generally composed by a septic tank as pretreatment, followed by soil infiltration field or sand filtration bed. The vertical drained sand filter provides the purification capacity thanks to the presence of a biomass in form of the biofilm. The dynamic of the biomass growth or the biofilm development is under the impact of filter materials’ nature. In this context, the objective of this work is to understand the mechanisms involved and especially the impact of medium in the functioning of the filtration reactor by comparing two types of packing materials: river sands and crushed aggregates. For this purpose, an experimental study is conducted with pilot unity composed by filtration reactors of 30cm of diameter and different packing thicknesses (15, 30 and 70cm). The reactors packed with two river sands and two crushed aggregates are fed with septic effluent with a volumic hydraulic charge of 12cm/day by 10 batches per day. Based on a characterization of materials, a study of purification performance with biochemical components monitoring of the total biomass and the extracellular matrix of the biofilm is realized by comparing two types of filter materials. The purification performance has presented similar efficiencies of particulates and organic matters removals by fine river sand and fine crushed aggregate. The nitrogen pollutants removals are more effective in the fine river sand which presents the finest granulometry with an alternative of aerobic and anoxic phases along the reactor depth and with a biomass more abundant. The distribution and the quality of the total biomass and the extracellular matrix differentiate between the river sand and the crushed aggregates. As the reference material, the fine river sand presented an earlier stabilization of total biomass growth with a less important production of extracellular exudates compared to the crushed aggregates. The origin of impacts brought by the crushed aggregates might be due to the higher fine particles content which created microenvironments poor in substrates or in oxygen and also due to a more heterogeneous mineralogy. The extracellular components of highest percentage in the biofilm of crushed aggregate are polysaccharides type substances

The objective of this investigation was to assess the potential of a titration bioassay i.e.: The Methanogenic Activity and Inhibition Analyser (MAlA), to determine the biodegradability of complex industrial effluents and wastewaters. Specifically, the project aimed to provide an alternative experimental method to the serum bottle method so that hazardous effluents can be pre-screened for treatment in under-utilised anaerobic digesters at sewage treatment plants in KwaZulu-Natal, South Africa. This study also aimed to provide a protocol and a simple mathematical model as experimental tools that could contribute to the development of future pre-screening studies. MAIA was used to conduct biodegradability and toxicity studies on semi-hazardous landfill leachate and textile size effluent. Thereafter, selected studies were repeated using a conventional screening method i.e.: serum bottle method. The investigation with MAlA revealed that both effluent substrates had potential for anaerobic treatment. However, the studies highlighted certain intrinsic limitations of the MAIA apparatus to effectively pre-screen complex substrates. The existing titrimetric system is too coarse to accurately track the biochemical pathways leading from the breakdown of complex compounds to methane gas production. Further, temperature interferences and gas phase diffusion limitations associated with the existing design make the assessment of activity difficult. The titrimetric method is comparable to the serum bottle method only if a qualitative assessment of toxicity and biodegradability is needed. However, the titrimetric method produces results in a much shorter period of time compared to the serum bottle method. Evaluated in this way the titrimetric method is the better alternative. However, the current system cannot challenge the reliability of the serum bottle method to provide good quantitative results. A mathematical model was developed which is much less detailed than the existing one provided by Remigi (2001). It comprises only two significant anaerobic processes namely hydrolysis and acetogenesis. Simulation trials have suggested that the model is a necessary and beneficial component ofthe titrimetric pre-screening protocol. This investigation has also led to the development of a more refined operating manual for MAIA.The manual provides a step-wise method for the preparation and conduction of pre-screening tests. Specifically, it highlights the need for a suitable biomass acclimation period and the importance ofnutrient use for better pre-screening assessments.
Thesis (M.Sc.Eng.)-University of Natal, Durban, 2003.

M. Tech. (Chemistry, Faculty of Applied and Computer Sciences), Vaal University of Technology.
Advanced oxidation processes (AOPs) are supposedly effective means for removal of low concentration of organic pollutants from waste water as compared to conventional treatment methods. However, TiO2 metal semiconductor is the most promising photocatalyst for degradation of organic pollutant under heterogeneous photocatalysis as compared to other metal semiconductors. Challenges such as aggregation in solution, low adsorptive ability for non-polar organic contaminants and recycling are limitations in application of TiO2 for commercial purposes. The other limitations of TiO2, is it only utilizes 4-6% of the solar energy reaching the earth's surface which is in the UV region and also rapid electron-hole recombination due its wide band gap. In this work, the limitations are overcome by synthesis of a new photocatalyst material and further applied on degradation of model organic contaminants. The first part of this work focused on preparation and characterization of photocatalyst material. The photocatalyst synthesized were characterized using X-ray diffraction (XRD), scanning electron microscopy (SEM), energy-dispersive X-ray analysis (EDAX), Fourier transform infrared spectroscopy (FTIR), thermal gravimetric analysis (TGA) and UV-VIS diffuse reflectance spectrophotometer (DRUV-VIS). Supporting characterization techniques revealed partly dispersion of TiO2 within the cavities of dealuminated Clinoptilolite (HCP). TiO2 exist as nanoparticles or clusters on the HCP surface ascribed to lower loading of TiO2. XRD analysis showed that the support material employed was mainly Clinoptilolite and absorption band of prepared photocatalyst was red-shifted into the visible region, with slight reduction in band gap of photocatalyst. The second part focused on adsorption and photocatalytic degradation of methyl orange solution (MO) conducted under UV-irradiation in the presence of TiO2/HCP. The influence of operational parameters on degradation efficiency of photocatalyst material on MO was carried out in this study. Parameters such as initial dye concentration, pH, calcination temperature, inorganic anions and peroxide concentration were varied during degradation activities of MO. Comparative degradation efficiency of TiO2/HCP, TiO2 and HCP were conducted on dye mixture (Methyl orange and Methylene Blue) under UV irradiation. Kinetic analysis employing Langmuir-Hinshelwood model on dependencies of organic contaminants degradation was also conducted at different operational parameters. The adsorption capacity of MO was highest in the presence of TiO2/HCP at lower loading, which is ascribed to good dispersion of TiO2 on HCP and increased surface area of dealuminated Clinoptilolite. The photocatalytic degradation of methyl orange in the presence of TiO2/HCP was optimized at low dye concentration (30 ppm), acidic condition (pH 4), and calcination temperature of 873 K. Nitrate ion of Sodium salt accelerates degradation activities on methyl orange as compared to other inorganic anions. Photocatalytic degradation of methyl orange was greatly enhanced upon addition of oxidant (H2O2) and the photocatalyst possessed good repeatability after 3 cycles. TiO2/HCP exhibit highest degradation activities, followed by HCP as compared to TiO2 during the degradation of dye mixture. The degradation of MO by the photocatalyst fits into pseudo-first order kinetic model, while for comparative analysis of photocatalyst on dye mixtures follows second order kinetic model.

M. Tech. Engineering : Chemical.
Evaluates the applicability of adsorption, biodegradation and hybrid adsorption and biodegradation system in treatment of wastewater containing melanoidin.Treatment of wastewater containing melanoidin through SAB process in fluidized bed bioreactor results in the best performance index as compared to adsorption and biodegradation processes undertaken singly. The synergies realized are more pronounced in fluidized bed bioreactor than in stirred tank system.

Industrial effluents with high concentrations of heavy metals are widespread pollutants of great concerns as they are known to be persistent and non-degradable. Continuous monitoring and treatment of the effluents become pertinent because of their impacts on wastewater treatment plants. The aim of this study is to determine the correlation between heavy metal pollution in water and the location of industries in order to ascertain the effectiveness of the municipal waste water treatment plant. Heavy metal identification and physico-chemical analysis were done using Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES) and multi-parameter probe respectively. Correlation coefficients of the measured values were done to investigate the effect of the industrial effluents on the treatment plants. Heavy metal resistant bacteria were identified and characterised by polymerase chain reaction and sequencing. Leeuwkuil wastewater treatment plants were effective in maintaining temperature, pH, and chemical oxygen demand within South Africa green drop and SAGG Standards whereas the purification plant was effective in maintaining the values of Cu, Zn, Al, temperature, BOD, COD, and TDS within the SANS and WHO standard for potable water. This findings indicated the need for the treatment plants to be reviewed.The industrial wastewater were identified as a point source of heavy metal pollution that influenced Leeuwkuil wastewater treatment plants and the purification plants in Vaal, Vereenining South Africa. Pseudomonas aeruginosa, Serratia marcescens, Bacillus sp. strain and Bacillus toyonensis that showed 100% similarity were found to be resistant to Al, Cu, Pb and Zn. These identified bacteria can be considered for further study in bioremediation.
Environmental Sciences
M. Sc. (Environmental Science)

Increase in magnitude of the global freshwater crisis together with the constantly changing demographics, hydrological variability and rapid urbanization will allow for continuous over exploitation of existing water resources, in an attempt to satisfy the rising socioeconomic demands. Increasing pressure on existing wastewater treatment plants, together with inefficient hygiene practices have exacerbated the nutrient and microbiological loads constantly entering surrounding water systems. This, coupled with the use of outdated guidelines has resulted, not only in an increase in waterborne related diseases but also an increase in waterborne-disease-related deaths. The current study investigated the physicochemical and microbiological quality of treated effluent from two independent wastewater treatment plants as well as their impact on the receiving watershed within Durban, South Africa over a one year period. Microbiological and physicochemical profiles were determined using standard methods whilst conventional PCR was used for the seasonal detection of human enteric viruses. Monthly variations were observed for all parameters with eight and six out of 12 month samples exhibiting increases in turbidity at the discharge point for the NWWTP and NGTW respectively, relative to before chlorination. Similarly, increases in nitrate and phosphate levels at the discharge point were also noted with the highest being recorded during December (215.23%) and September (12.21%) respectively. Temperature profiles ranged between 12 – 26 °C and 12.7 – 26 °C for the NWWTP and receiving Umgeni River whilst for the NGTW and receiving Aller River, it ranged between 16.5 – 26 °C and 12 – 25.7 °C respectively. Seasonal averages revealed relatively high COD values downstream of the Umgeni River during winter (263.22 mg/l) and spring (177.93 mg/l). Eight out of twelve samples exhibited increases in turbidity at the discharge point for the NWWTP with the highest values obtained during April (76.43 NTU). Significant positive correlations (p ≤ 0.05) were observed upstream and downstream of the Umgeni River between temperature and BOD (r = 0.624); turbidity (r = 0.537); TDS (r = 0.437); TSS (r = 0.554) and DO (r = 0.516). Percentage reduction of bacterial indicators at the discharge point ranged between 0.52 – 100% and 41.56 – 100% across the sampling period for the NWWTP and NGTW, respectively. Treated effluent from both plants did not meet the required guidelines, with a 100% reduction in the faecal coliform load being detected only during October 2012 for both plants. In addition, higher levels of indicator bacteia were observed at the discharge point for the NWWTP during February 2013 with observed counts (in CFU/ml) as high as 12.27 x 103; 6.61 x 103; 2.99 x103; 1.6 x 103 and 1.17 x103 for total coliforms, E.coli, faecal coliforms, faecal streptococci and enterococci, respectively. Similarly, higher levels of both somatic and F-RNA bacteriophages were detected during April (106.67 PFU/ml), May (309.33 PFU/ml). June (346.67 PFU/ml) and August (126.67 PFU/ml) compared to samples collected before chlorination for the NWWTP. Enteroviruses were detected in 100% of unchlorinated final effluent samples, 87.5% of chlorinated final effluent and 93.75% of receiving river samples whilst human adenoviruses were detected in 50% of final effluent samples before chlorination, 62.5% in samples collected at the discharge point and 62.5% of river water samples. This study revealed that whilst the independent treatment plants monitored, exhibited effluent qualities that met acceptable standards for some parameters such as pH and temperature, the effluent quality fell short of other standard requirements. Ensuring efficient surveillance and management of existing treatment plants coupled with guideline revision and monitoring compliance is imperative in preventing further risk of pollution to both the environment and human health.
Thesis (M.Sc.)-University of KwaZulu-Natal, Durban, 2013.

M.Tech.
The removal and recovery of heavy metals from effluents has been a subject of significant importance due the negative impact these toxic metals have on human health and the environment as a result of water and soil pollution. Precipitation is the mostly widely used wastewater treatment method because it is the most economical and easier to implement and operate on a large scale. However, traditional precipitation methods using lime, sulfides or hydroxides recover metals in the form of a sludge which is not reusable and has to be disposed in landfills creating a potential environmental hazard and resulting in loss of valuable minerals. The current focus in effluent treatment is now on the recovery and re-use of these heavy metals rather than removal and disposal. This study investigated the use of hydrazine as a reducing agent to remove and recover Ni2+, Cu2+, Co2+ and Fe2+ from effluent by reduction crystallization. In this process chemically reduced aqueous metal ions were plated on to a base substrate (nickel powder) with no electrical current required for deposition. A feasibility study was carried out to test the efficiency and find the optimum operating conditions for this method and generate an understanding of the chemical and particulate process occurring. The results obtained indicate that hydrazine is an effective reducing agent for removal and crystallization of Ni2+, Cu2+, Co2+ and Fe2+/ Fe3+ into their elemental states with nickel powder as a seeding material. Over 99 % of metals were removed from the effluent in all the systems (Ni-only, Ni-Cu, Ni-Fe and Ni-Fe). Breakage, aggregation and molecular growth were identified as the predominant mechanisms occurring during the reduction crystallization process in Ni-only, Ni-Cu, Ni- Co systems and there was evidence of nucleation in Ni-Fe solution. These finding were confirmed by analysing the scanning electron micrographs of the powder obtained. A nearly spherical structure powder with wide distribution in particle size and evidence of fragmentation was obtained in all the experimental runs. vii The residual concentrations obtained were far below the required limit for effluent discharge into sewer where 20 mg/L Ni, 20 mg/L Cu and 20 mg/L Fe and the total metal concentration of 50 mg/L for Fe, Cr, Cu, Ni, Zn and Cd is stipulated. Reduction crystallization using hydrazine as a reducing agent can be utilized for controlling environmental pollution and eliminating hazardous metals from the environment.

The presence of heavy metal ions in waste streams is one of the most pervasive environmental issues of present times. A rotating biological contactor (RBC) was used to investigate the potential capacity of microbial biofilms in remediation of the metal ion species from a mixed metal contaminated effluent solution containing Cr+3 , Pb+2 and Cu+2 , each at a concentration of 200 mg r1 • In the first part of this study the effectiveness of various support materials for the development of microbial biofilms capable of removing heavy metals from a synthetic effluent was investigated. EDX analysis showed that none of the support matrices investigated, viz. gravel, polyester batting and sand, adsorbed metal ions on their surfaces; hence, metal adsorption was due purely to microbial activities. The biofilms attached more firmly and uniformly to polyester batting than to gravel and sand. The characteristics of polyester batting which made it a superior support matrix were its surface roughness and porous hydrophilic nature, which provided a larger surface area for the adhesion of microorganisms and attraction of nutrients during the biofilm development process. The selective accumulation of metal ion specIes by various microbial populations grown as biofilm using polyester batting as support matrix in separate compartments of a single-stage RBC bioreactor was examined. Lead ions were readily accumulated by almost all the microbial biofilms tested. Fungus-dominated biofilms selectively accumulated chromium ions whereas biofilms comprising mainly bacteria more readily accumulated copper ions from the mixed metal contaminated effluent solution. However, where interactions between the bacterial and fungal components were encouraged the mechanical stability of the biofilms was enhanced so that large amounts of all three metal ion species were removed by this biofilm. The combined effect of a series of bench-scale columns containing liquid humic acid and a three stage RBC bioreactor on the removal of metal ion species from a mixed metal contaminated effluent was investigated. After seven days of treatment the combined system had removed approximately 99% of the Cr+3, 98% of the Pb+2 and 90% of the Cu+2 ions from the mixed metal contaminated synthetic effluent. Complexation of the metal ions with humic acid was the predominant factor accounting for approximately 68-86% Cr+3 , 70-86% Pb+2 and 53-73% Cu+2 removal levels within the columns. A large proportion of the remaining Cr+3 and Pb+2, but not of the Cu+2, was removed in compartment 1 of the RBC. This suggested that the presence of the former two metals in solution might have reduced the removal of the Cu+2 ions from the system. The removal of substantially large amounts of the competing ions chromium and lead during the initial stages of the treatment process meant that copper was successfully taken up in the second and third RBC compartments. Hence, the economy of the treatment process was improved as larger quantities of the metal ions were removed in a shorter period of time than was possible when using the individual treatments (humic acid-metal complexation and biofilm adsorption) separately. More than 75%,92% and 86% of the adsorbed Cr+3 , Pb+2 and Cu+2 ions, respectively, were recovered from the three RBC bioreactor compartments following repeated washing of the biofilms with 0.1 M HCI. This relatively easy desorption suggested that the metal ions were simply adsorbed onto the surfaces of the biofilm cells rather than being taken into the cytoplasm of the cells.
Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2004.

A 2000 mm long saturated laboratory soil column was used to simulate soil aquifer treatment under saturated conditions to assess the removal of chemical and biochemical oxygen demand (COD and BOD), dissolved organic carbon (DOC), nitrogen and phosphate, using high strength artificial wastewater. The removal rates were determined under a combination of constant hydraulic loading rates (HLR) and variable COD concentrations as well as variable HLR under a constant COD. Within the range of COD concentrations considered (42 mg L(-)(1)-135 mg L(-)(1)) it was found that at fixed hydraulic loading rate, a decrease in the influent concentrations of dissolved organic carbon (DOC), biochemical oxygen demand (BOD), total nitrogen and phosphate improved their removal efficiencies. At the high COD concentrations applied residence times influenced the redox conditions in the soil column. Long residence times were detrimental to the removal process for COD, BOD and DOC as anoxic processes and sulphate reduction played an important role as electron acceptors. It was found that total COD mass loading within the range of 911 mg d(-)(1)-1780 mg d(-)(1) applied as low COD wastewater infiltrated coupled with short residence times would provide better effluent quality than the same mass applied as a COD with higher concentration at long residence times. The opposite was true for organic nitrogen where relatively high concentrations coupled with long residence time gave better removal efficiency.

Inadequately treated wastewater effluent is harmful to the receiving aquatic environment. Water-borne chemicals and microbial pathogens pose a health risk to anyone living downstream from sewage treatment facilities. This study assessed the effluent from a package plant with a design capacity of 48kℓ/24 hours, servicing 12 household units and a restaurant in Mogale City. Over a 12 month period, fortnightly water samples were collected from ten selected sites including two boreholes, a river and two dams. Standard parameters including physical (pH, EC, temperature, DO and SS), chemical (nutrient concentration) and biological (bacterial counts) were analysed using handheld meters, standard membrane filter techniques and colorimetric methods. One borehole was affected by pathogen and nitrate runoff from an adjacent poultry farm. If regularly monitored, the package plant effectively removed microbes (most samples contained 0 cfu/100mℓ) but above limit COD, ammonia and phosphate was released in the effluent (with maximum values of 322 mg/ℓ, 42.52 mg/ℓ and 7.18 mg/ℓ, respectively). Generally, river and dam water at the site was of good quality.
Environmental Science
M. Sc. (Environmental Science)