Polluted waters, arising from extensive past and ongoing mining operations in South Africa, pose serious environmental threats to the limited fresh water resource. The long time periods, of decades to centuries, over which decanting mine waters may be expected to flow raises additional concerns about the sustainability of these resources. Responses to the problem have thus increasingly been directed towards the long-term sustainability of mine water treatment technologies (MWTT) as a critical indicator in both their research and development, and application. Bioprocess treatments have been considered in this regard and, among these, the Rhodes BioSURE Process has been investigated in preliminary studies using complex organic carbon wastes as the carbon source and electron donor for the central sulphate reduction unit operation. Although both the mining industry and the related statutory/regulatory authority in South Africa share public commitment to sustainability in the treatment of mine waters, no systematic mechanism has emerged to enable the application of sustainability thinking as a guiding principle in the selection and application of MWTTs, nor in the research and development undertaking. This study undertook the development of a Sustainability Indicator Framework in order to provide a systematic basis for the incorporation of sustainability objectives in MWTT bioprocess development, and specifically to use this framework as an input to the investigation of the scaleup development of the Rhodes BioSURE Process. In the development of the MWTT Sustainability Indicator Framework, an initial survey of industry thinking in this area was undertaken and, based on these outcomes, a detailed questionnaire methodology was developed in order to identify and quantify critical sustainability indicators. These included analysis of environmental, economic, social and technical indicators used in sustainability accounting practice in the industry. Statutory/regulatory sustainability targets in the same categories were derived from State of the Environment Reports (SoER) from Provincial authorities where mining is undertaken in South Africa. A synthesis of industry and SoER values was derived from weighted averages and the Sustainability Indicator Framework based on these outcomes. A Conceptual Decision-Support System, to guide the selection and development of MWTTs, was proposed and also based on these results. In the development of the Rhodes BioSURE Process the use of primary sludge (PS) had been investigated as a potential complex carbon and electron donor source. In this regard the utility operator, and sewage treatment process infrastructure, was identified as potentially meeting aspects of the sustainability objectives identified for MWTT application development. Both the Sustainability Indicator Framework and the Conceptual Decision-Support System provided inputs in the formulation of the experimental programme relating to the scale-up development of the Rhodes BioSURE Process. Based on these outcomes, a series of single- and multi-stage reactor configuration, optimisation and enzymology studies were undertaken at bench-, pilot- and technical-scale operations. These units were operated at hydraulic retention times (HRT) ranging between 22 to 72 hours and at chemical oxygen demand to sulphate ratios (COD:SO[subscript 4]) ranging between 1:1 to 2:1. Studies undertaken in fed-batch, bench-scale reactors confirmed the preliminary feasibility of using established sewage treatment infrastructure as a replacement for novel reactor configurations that had been used in the initial studies. The results further indicated that the hydrolysis of PS occurred at different rates under biosulphidogenic conditions in the different reactor configurations investigated. Scale-up of these findings in multi-stage pilot- (7.4m[superscript 3]) and technical-scale plants (680m[superscript 3]) showed comparable performances between the unit operations in terms of SO[subscript 4] and COD removal. These results indicated no apparent advantages in the uncoupling of hydrolysis and sulphate reduction in separate unit operations as had been suggested in previous studies. Scale-down/scale-up studies were undertaken in a continuously fed single-stage reactor configuration and showed that the process could be effectively operated in this way. Previous proposals that chemical and biological gradients established in the sludge bed of the Recycling Sludge Bed Reactor (RSBR) exercised an influence on the rates of substrate hydrolysis were investigated and the relative activity of α- and β-glucosidase and protease enzymes was measured. Results provided additional support for this hypothesis and it was shown that enzyme assay may also provide a useful tool in process development and monitoring studies. While sulphide recovery, following the sulphate reduction step in the BioSURE Process, was not investigated as a component of this study, the treatment of final effluent or waste spills was identified as an important sustainability requirement given the toxicity of sulphide to human and ecosystem environments. A conventional trickle filter reactor system was evaluated for this purpose and showed close to 100% oxidation to sulphate in a short contact time operating regime. Although residual COD removal was low at ~20% of influent, it is considered that high rate recycle biofilter operation could achieve the COD discharge standard of 75 mg/l. The results of the above studies provided inputs into the design, construction and commissioning of the first full-scale commercial application of the Rhodes BioSURE Process for mine wastewater treatment using sewage sludge as the carbon and electron donor source. An adjacent mine and sewage works have been linked by pipeline and an operational capacity of 10 Ml/day water treated has been established with sulphate reduced from ~1300mg/l to <200mg/l. These developments constitute a novel contribution in the mine waste water treatment field.

An investigation of the biodegradation of complex hydrocarbon wastes, with emphasis on chlorinated aromatic compounds, in an anaerobic/aerobic bioprocess environment was made. A reactor configuration was developed consisting of linked anaerobic and aerobic reactors which served as the model for a proposed bioremediation strategy targeting subterranean soil/sediment/aquifer chlorinated phenol-contaminated environments. Here oxygen is frequently limited and sulphate is readily available, as occurs especially in marine sediment and intertidal habitats. In the anaerobic system the successful transformation and mobilization of the model contaminant, 2,4,6-trichlorophenol, was shown to rely on reductive dechlorination by a sulphate-reducing dependent dechlororespiring co-culture. This was followed in the aerobic system by degradation of the pollutant and its metabolites, 2,4-dichlorophenol, 4-chlorophenol and phenol, by immobilized white-rot fungi.The strategy was initially investigated separately in laboratory bench- and intermediate scale reactors whereafter reactors were linked to simulate the integrated biodegradation strategy. The application of the fungal reactor to treat an actual waste stream by degrading complex mixtures of hydrocarbons in a waste oil recycling effluent was also investigated. The mineralization of phenol and 2,4,6-TCP by immobilized fungal cultures was studied in pinewood chip and foam glass bead-packed trickling reactors. The reactors were operated in sequencing batch format. Removal efficiency increased over time and elevated influent phenol and TCP (800 and 85 mg.L⁻¹) concentrations were degraded by > 98 % in 24 – 30 h batch cycles. Comparable performance between the packing materials was shown. Uptake by the packing was negligible and stripping of compounds induced by aeration had a minimal effect on biodegradation efficiency. Reactor performances are discussed in relation to sequencing batch operation and nutrient requirements necessary to sustain fungal activity in inert vs. organic material packed systems. It was shown that a co-culture consisting of sulphate-reducing and dechlororespiring bacteria established in fed-batch and soil flasks, as well as pine chip-packed fluidized bed reactors. Results showed reductive dechlorination of 2,4,6-TCP to be in strict dependence on the activity of the sulphate-reducing population, sulphate and lactate concentrations. Transformation to 2,4-DCP, 4-CP and phenol was enhanced in sulphate deficient conditions. Dechlororespiring activity was found to be dependent on the fermentative activity of sulphate-reducing bacteria, and the culture was also shown to mobilize and dechlorinate TCP in soils contaminated with the pollutant. Linking the systems achieved degradation of the compound by > 99 % through fungal mineralization of metabolites produced in the dechlororespiring stage of the system. pH correction to the anaerobic reactor was found to be necessary since acidic effluent from the fungal reactor inhibited sulphate reduction and dechlorination. The fungal reactor system was evaluated at intermediate-scale using a complex waste oil recycling effluent. Substantial COD reduction (> 96 % in 48 h batch cycles) and removal of specific effluent hydrocarbon components was shown in diluted, undiluted (COD > 37 g.L⁻¹) and 2,4,6-TCP-spiked effluents. Industrial application of the fungal reactor was evaluated in a 14 m³ pilot plant operated on-site at a waste oil processing plant.

Water is an essential element in all aspects of life and is vital for both domestic and industrial purposes regarding both the quality and quantity thereof. Similar to many other drought stricken countries, South Africa requires water for the socio-economic growth of the country, yet is faced with the problem of maintaining the quality of its drinking water as well as protecting the dwindling supplies. In an attempt to prevent the deterioration of South African water supplies the treatment, purification and recycling of industrial and mining waste water has recently become of prime importance. Many industrial and mining waste waters contain heavy metals in toxic quantities. The conventional processes that have been used till recently to address this problem, are often expensive or contain chemical agents which compound the environmental problem. As an alternative biological methods of metal accumulation appear to offer an economic and efficient alternative to these methods. An advantage to the South African scenario is the commercial production of the yeast, S. cerevisiae as a readily inexpensive by-product from some fermentation industries, Yeast cells, and in particular S. cerevisiae have proven to be capable of accumulating heavy metals, and therefore exhibit potential application in the bioremediation of waste water. The aim of this project was twofold. The initial part of this work attempted to define the mechanisms of metal accumulation by the yeast cells and cellular components. The information obtained from these initial studies provided a data base required for the development of a bioremediation system. Initial contact with the metal ions occurs at the wall interface of the yeast cell. Metal accumulation appears to be a function of all the cell wall components. The isolated cell wall components are better metal chelators then the intact cell walls. An apparent affinity series of mannan > chitin> glucan > intact cell walls exists. However, these components differ in their affinities for metal ions. Storage of metal ions within the cell occurs predominantly in the vacuole. The present study concluded that metal accumulation by the vacuole could be related to size. Metal accumulation occurred in the order of Cu2+ > Co2+ > Cd2+ with a corresponding decrease in atomic radii of Cd2+ > C02+ > Cu2+. Vacuolar ion deposition occurs at an early stage during the internalization of metal ions within the yeast cells. At the onset of vacuolar saturation, depositions of metal ions as granules within the cytosol occurs. In the presence of heavy metal cations viable yeast cells can be shown to exhibit two types of cellular responses. Uptake of Cu2+ and Cd2+ causes the loss of intracellular physiological cations from within the yeast cell. In comparison, uptake of Co2+ into the cell does not have this effect. All three heavy metal cations initiate plasma cell membrane permeability, thus the Cu2+ and Cd2+ induced loss of the intracellular cations, occurs. ~ a result of ion-exchange mechanisms and not due to cation leakage brought about by membrane permeabilization. Uptake of heavy metals by viable yeasts appears to be generally non-selective though the amount of metals accumulated are largely affected by the ratio of ambient metal concentration to biomass quantity. In addition, the energy dependent nature of internalization necessitates the availability of an external energy source for metal uptake by viable yeast cells. For these reasons metal removal from industrial waste water was investigated using non-viable biomass. By immobilizing the yeast cells additional mechanical integrity and stability was conferred apon the biomass. The three types of biomass preparations developed in this study, viz. polyvinyl alcohol (PV A) Na-alginate, PV A Na-orthophosphate and alkali treated polyethylenimine (PEI):glutaraldehyde (GA) biomass pellets, all fulfilled the necessary physical requirements. However, the superior metal accumulating properties of the PEI:GA biomass determined its selection as a biosorbent for bioremediation purposes. Biosorption of heavy metals by PEI:GA biomass is of a competitive nature, with the amount of metal accumulated influenced by the availability of the metal ions. This availability is largely determined by the solution pH. At low pH values the affinity of the biomass for metals decreases, whilst enhanced metal biosorption occurs at higher pHs, ego pH 4.5 - 6.0. PEI:GA biomass pellets can be implemented -as a biosorbent for the bi9remediaiton of high concentration, low-volume metal containing industrial waste. Several options regarding the bioremediation system are available. Depending on the concentration of the metals in the effluent, the bioremediation process can either be used independently or as part of a biphasic remediation system for the treatment of waste water. Initial phase chemical modification may be required, whilst two types of biological systems can be implemented as 'part of the second phase. The PEI:GA biomass can either be contained within continuous-flow fixed bed tanks or continuous-flow stirred bioreactor tanks. Due to the simplicity of the process and the ease with which scale-up is facilitated, the second type of system shows greater application potential for the treatment of this type of industrial waste water than the fixed-bed systems.

The hydrolysis of complex organic heteropolymers contained in municipal wastewater to simpler monomers by extracellular hydrolytic enzymes is generally considered the rate-limiting step of the biodegradation process. Previous studies of the Recycling Sludge Bed Reactor (RSBR) revealed that the hydrolysis of complex particulate organics, such as those contained in primary sludge (PS), was enhanced under anaerobic biosulphidogenic conditions. Although the mechanism was not fully understood, it appeared to involve the interaction of sulfide and sludge flocs. The current study was conducted using a 3500 ml laboratory-scale RSBR fed sieved PS at a loading rate of 0.5 kg COD/m³.day and an initial chemical oxygen demand (COD) to sulfate ratio (COD:SO₄) of 1:1. There was no significant accumulation of undigested sludge in the reactor over the 60-day experimental period and the quantity of SO₄ reduced indicated that the yield of soluble products from PS was at least as high as those reported previously for this system (> 50%). In the current study, the specific activities of a range of extracellular hydrolytic enzymes (L-alanine aminopeptidase, L-leucine aminopeptidase, arylsulphatase, α-glucosidase, β- glucosidase, protease and lipase) were monitored in a sulfide gradient within a biosulphidogenic RSBR. Data obtained indicated that the specific enzymatic activities increased with the depth of the RSBR and also correlated with a number of the physicochemical parameters including sulfide, alkalinity and sulfate. The activities of α- glucosidase and β-glucosidase were higher than that of the other enzymes studied. Lipase activity was relatively low and studies conducted on the enzyme-enzyme interaction using specific enzyme inhibitors indicated that lipases were probably being digested by the proteases. Further studies to determine the impact of sulfide on the enzymes, showed an increase in the enzyme activity with increasing sulfide concentration. Possible direct affects were investigated by looking for changes in the Michaelis constant (Km) and the maximal velocity (Vmax) of the crude enzymes with varying sulfide concentrations (250, 400 and 500 mg/l) using natural and synthetic substrates. The results showed no significant difference in both the Km and the Vmax for any of the hydrolytic enzymes except for the protease. The latter showed a statistically significant increase in the Km with increasing sulfide concentration. Although this indicated a direct interaction, this difference was not large enough to be of biochemical significance and was consequently not solely responsible for the enhanced hydrolysis observed in the RSBR. Investigation into the floc characteristics indicated that the biosulphidogenic RSBR flocs were generally small in size and became more dendritic with the depth of the RSBR. Based on the above data, the previously proposed descriptive models of enhanced hydrolysis of particulate organic matter in a biosulphidogenic RSBR has been revised. It is thought that the effect of sulfide on the hydrolysis step is primarily indirect and that the reduction in floc size and alteration of the floc shape to a more dendritic form is central to the success of the process.

No abstract available. / Dissertation (MSc (Water Utilization))--University of Pretoria, 2006. / Chemical Engineering / unrestricted

Sewage-holding tanks aboard recreational boats store human wastes, thereby preventing the direct discharge of wastewater to the aquatic environment. Water-conserving toilets and limited holding tank volumes produce a highly concentrated waste that must be periodically dumped to a wastewater treatment system. Prior to disposal, many boat operators add commercial preparations to control odors produced in their chemical toilets and holding tanks. The objective of this study was to determine the effects of three holding-tank chemicals on anaerobic wastewater treatment. Specifically, septic-tank performance with respect to effluent total suspended solids (TSS) and chemical oxygen demand (COD) was evaluated. Potential drain-field failure was the concern that led to the selection of TSS and COD. Drain-field failure could result from high solids carry-over or from a high concentration of COD in the effluent which would promote excessive bio-mat growth and clog the system. Laboratory septic tanks were constructed and operated for this evaluation. Methanol, paraformaldehyde and formaldehyde were each listed as an active ingredient in one of three chemical compounds used by recreational boat owners to deodorize sewage-holding tanks. septic-tank effluent TSS concentrations were not adversely effected by the shockloading with wastewater containing these chemicals. Concentrations expected to be achieved by dilution (20 and 50 percent of the recommended additive dose) resulted in septic-tank effluent COD within an acceptable range, which was determined by operation of a control system. Wastewaters containing these concentrations were not detrimental to the septic-tank treatment system. However, the full manufacturers' recommended dose of the odor control chemicals disrupted the system's ability to degrade COD. At full strength, the para formaldehyde and formaldehyde deodorants were particularly detrimental; no recovery occurred after the two-day shock-dose was completed. / Master of Science

The objective of this study was to examine the influence of nutrients on the biological phosphorus removal process at high acetate concentrations. It was an extension of studies conducted by Randall and Chapin (1994), who found that industrial wastewater with high concentrations of acetate were able to inhibit the biological phosphorus removal process. Two bench-scale pilot plants were operated under controlled conditions that included synthetic wastewater as feed. The acetic acid concentrations in the feed of one system was increased in steps from 200 to 800 mg/L while the acetic acid concentrations in the feed of the other system was constantly held at 200 mg/L. Sludge from both systems was used for batch tests determining the kinetics of phosphorus release and uptake and poly-β-hydroxybutyric acid synthesis. Furthermore, the influence of various nutrients were examined during these batch tests. The results of this study confirmed the observations of Randall and Chapin (1994). High concentrations (600 mg/L) of acetic acid did inhibit the biological phosphorus removal process; however, this inhibition could be countered by adding calcium into the feed. The reactions of phosphorus release and uptake are described by first order kinetics. / Master of Science

Thesis (MSc)--University of Stellenbosch, 2001. / ENGLISH ABSTRACT: South Africa is one of the world's major coal producers, resulting in the second highest foreign exchange earner for South Africa. However, the mining industry contributes negatively to (ground) water pollution, due to the formation of acid mine drainage (AMD). AMD originates from the bacterial oxidation (Thiobacillus ferrooxidans) of pyrite (FeS) and contains high levels of sulphate and metals. Sulphate rich waters can be treated applying the biological sulphate removal technology. This study concentrated on biologically removing sulphate from synthetic feed- and mine water, using the single-stage completely-mixed reactor system. The advantage of using this reactor system is that except for removing sulphate from about 2000 to less than 200 mg/t', it can also partly biologically remove the formed sulphides. It was established that both ethanol and sugar can be used, as the carbon and energy source, however ethanol is more cost effective than sugar. Ethanol dosage and Hydraulic Retention Time (HRT) studies were undertaken to investigate at what concentration, the highest sulphate and sulphide removal rates were achieved. It was found that the highest sulphate reduction rates were obtained when using 1mf ethanol/f feed and that the removal rates were dependent on the HRT: the lower the HRT, the higher the sulphate reduction rate. The highest sulphide oxidation rate was achieved at the HRT of 6 h. It was, furthermore shown that the single stage completely-mixed reactor system could successfully be used to remove sulphate from Schoongezicht mine effluent, not only removing the sulphate, but also most of the metals, thereby increasing the mine effluent pH from 2.5 to 7. The conclusion of this study was that a completely-mixed reactor system, as described in this thesis, can successfully be applied to treating acid mine drainage using ethanol (1 m.e etanol/f feed water) as the carbon and energy source at a hydraulic retention time as low as 4 hours. This technology has great potential for pilot- and full-scale treatment of sulphate rich effluents such as acid mine drainage. / AFRIKAANSE OPSOMMING: Suid Afrika is een van die vemaamste steenkool produseerders in die wereld, terwyl die uitvoer van steenkool die land se tweede hoogste verdiener is van buitelandse valuta. Ongelukkig dra hierdie industrie ook by tot die besoedeling van (grond) water, veral vanwee die vorrning van suur myn afloop. Bakteriese oksidasie (deur Thiobacillus ferrooxidansy van piried (FeS) is hoofsaaklik verandwoordelik vir die vorrning van suur myn afloop bevattende hoe konsentrasies van sulfaat en metale. . Die toepassing van biologiese sulfaatverwyderingsprosesse vir die behandeling van sulfaatryke waters is vroeer gedemonstreer. Die doel van hierdie studie was om 'n enkel-stadium reaktor met volledige vermenging te evalueer en te optimiseer om toegepas te word vir die biologiese verwydering van sulfaat vanuit sinteties bereide, sowel as mynwater. Hierdie reaktor is in staat om sulfaat te verwyder vanaf vlakke van ~ 2000 tot minder as 200 mg/P. 'n Verdere voordeel gepaard met die gebruik van hierdie reaktor is dat die sulfied wat gevorm word tydens sulfaatreduksie, gedeeltelik verwyder word deur die oksidasie daarvan na So. Die resultate wat behaal is in hierdie studie het aangedui dat beide etanol en suiker gebruik kan word as die koolstof en energiebron, terwyl etanol meer koste-effektief aangewend kon word. In teenstelling was metanol nie 'n geskikte koolstofbron vir sulfaatverwydering nie. Eksperimente is daarvolgens uitgevoer om toestande van optimum etanoldosering en hidroliese retensietyd (HRT) vir maksimum sulfaat- en sulfiedverwydering te bepaaJ. Die hoogste reduksie tempo's was verkry met 'n toediening van 1 mP etanol/f invloei, en die effektiwiteit van verwydering was afhanklik van HRT. Hoe laer die HRT, hoe hoer die tempo van sulfaatverwydering. Die beste sulfaatverwyderingstempo was behaal teen 'n HRT van 6 uur. Die resultate het verder aangetoon dat die enkel-stadium reaktor met volledige vermenging in staat was om sulfaat effektief te verwyder, en die pH te verhoog vanaf na 2.5 tot 7, in mynuitvloeisels van 'n plaaslike steenkoolmyn. Die gevolgtrekking uit hierdie werk is dat 'n volledig-gemengde reaktorstelsel, soos beskryf in die huidige studie, geskik is vir die suksesvolle behandeling van suur mynafloopwater met die gebruik van etanol (l mflP toevoerwater) as koolstof- en energiebron by 'n hidrouliese retensietyd tot so laag as 4 uur. Die tegnologie het groot toepassingspotensiaal vir volskaalse behandeling van sulfaatryke afloopwaters soos by. suur mynafloop.

A research report submitted to the Faculty of Engineering and the Built Environment, University of the Witwatersrand, Johannesburg, in Partial Fulfilment of the requirements for the degree of Masters of Science in Engineering, 2017 / Access to good quality drinking water is essential for the maintenance of public health. To guarantee a steady supply of good quality water, water treatments plants are designed to provide potable water that meets national and, where necessary, local water quality standards. While the protection of natural water resources against pollution, and proper treatment of water at treatment plants are both crucial to the provision of safe drinking water, the reality is that the quality of treated water can degrade during distribution. Microbial proliferation within distribution systems can cause problems such as unpleasant tastes and odours as well as the proliferation of pathogenic microorganisms. For most utilities, it is biofilm that grows on pipe surfaces that act as permanent inocula continuously inoculating the bulk water as it flows through the distribution system. Distribution system biofilm growth and the resulting impact on water quality can be minimized by various treatment processes, designed to remove biodegradable organic matter (BOM) from the water. The removal of BOM is of great importance to water utilities because it eliminates bacterial regrowth and the many associated water quality issues. Hence, the spatial and temporal mapping of biodegradable organic carbon (BDOC) offers water utilities an effective strategy in managing the BOM in the distribution system. This research is aimed at evaluating the applicability of BOM measurement protocols to monitoring biostability and biofilm formation potential within a drinking water distribution system (DWDS). This study specifically investigated the efficacy of a simplified version of the high-density BDOC test as a protocol for monitoring BDOC in finished water. The high-density BDOC protocol was found to be a more streamlined approach in contrast to the assimilable organic carbon (AOC), and provides a suitable monitoring mechanism for lowering biofilm formation potential in DWDSs. / CK2018

University of Macau / Faculty of Science and Technology / Department of Civil and Environmental Engineering