Studies involving solids-stabilised foams have been limited and few have focused on the benefits of complex systems such as those involving mixtures of more than one surfactant. Little is known about the effectiveness of using mixed surfactant systems as foam stabilisers at the bulk level. The purpose of this project was to gain further understanding and insight into foam stability, on a bulk scale, in the absence, and in the presence, of solid colloidal particles, and for systems involving single or mixed surfactants. Foams were produced using sodium dodecyl sulphate (SDS), dodecanoic acid (DA), or a mixture of both surfactants at varying molar ratios. The surface tension at a given concentration, the limiting surface tension (at high concentrations), the critical micelle concentration, foamability and foam stability were all significantly affected by the molar ratio of SDS to DA. The initial pH of mixed surfactant solutions played a role in the shift of surface tension and CMC. The higher pH values, the lower surface activity, therefore resulted in higher surface tension. At a given pH, however, the mixed surfactant solutions with higher molar ratio of SDS to DA appeared to have lower CMC. The foam stabilising ability of colloidal dispersions of four hydrous metal oxides, namely hydrous iron oxide (formed by hydrolysis of Fe(III) solutions and referred to as HFO), hydrous zinc oxide (formed by hydrolysis of Zn(II) solutions and referred to as HZO), hydrous chromium oxide (formed by hydrolysis of Cr(III) solutions and referred to as HCO), and hydrous nickel oxide (formed by hydrolysis of Ni(II) solutions and referred to as HNO) were studied at varying concentrations. Generally foam stability increased as the solid concentration increased. Foams stabilised by HNO were found to be the most stable. Foams stabilised by HFO were found to be unstable regardless of the solid concentration. It is believed that the instability of such foams is primarily due to the large aggregated size of HFO flocs. The aggregate size of hydrous metal oxides was influenced by the concentration of NaOH used to hydrolyse the metal ion solution, and by sonication treatment immediately following solid formation. However, the final pH of the colloidal dispersions did not significantly change the aggregate size. Solids stabilised foams are believed to be highly dependent on the state of hydrophobicity of the solids used, and this is in turn controlled by adjustment of the pH dependent surface charge and potential. Electrophoretic mobility is a commonly used tool to probe the potential near the surface and was used in this thesis to determine the affect of surfactant adsorption (particularly from solutions containing mixtures of SDS and DA) on surface properties of the solids. The electrophoretic mobility of all hydrous metal oxide aggregates decreased as the concentration of SDS/DA increased. Specific adsorption was evident in all cases and resulted in charge reversal for most cases. Electrophoretic mobility data for surfactant adsorption, as a function of total surfactant concentration, was consistent with a three-stage model of surfactant adsorption involving (1) electrostatic adsorption, (2) cooperative adsorption and (3) surface saturation. The influence of surfactant adsorption on electrophoretic mobility was found to be consistent with models requiring the stabilising solids to be in a controlled state of flocculation, where the zeta potential (as probed by electrophoretic mobility) must be sufficiently high to prevent total flocculation of the solid and thus collapse of the foam, but sufficiently low that the solids have some degree of hydrophobicity such that they prefer to be only partially wetted (and thus reside at the air/water interface). The total percentage adsorption of SDS/DA surfactant mixtures on both hydrous iron oxide and hydrous nickel oxide was found to be independent of time. However, the proportion of adsorption due to SDS and DA was dependent on time. Moreover, the initial and final ratio of SDS to DA adsorption did not reflect their ratio in solution. SDS adsorbed, initially, to a greater extent that would be predicted from its solution concentration, but this trend was reversed after a period of time.

Submitted in partial fulfillment of academic requirements for the degree of Masters of Technology: Department of Chemical Engineering, Durban University of Technology, 2009. / The primary objective of this project was to establish a screening protocol which could be used to access high strength/toxic effluent for toxicity and degradability prior to being disposed in wastewater treatment works. The serum bottle method (materials and method section) is simple, makes use of small glass vials (125 mℓ-volume were used in this research) which do not require any stirring nor feeding device or other engineered tool: a serum bottle is sealed immediately after all components are poured inside and thereafter conducted in a batch mode and occasionally shaken to ensure adequate homogenisation of the components. The only variables which are regularly measured are the volume of biogas produced and gas composition. The two assays, originally developed by Owen et al. (1979) to address the toxicity and the biodegradability have been combined in a single test called AAT, Anaerobic Activity Test, which enables one to assess simultaneously the inhibitory effect on the methanogenic biomass and the biodegradability of the test material as well as the ability of the biomass to adapt to the test material and therefore to overcome the initial inhibition. The screening protocol is illustrated in Annexure A. The protocol consists of a sequence of assays which employ the serum bottle methodology. A first step of the procedure is aimed at rapidly estimating whether the effluent is potentially toxic to the methanogenic biomass and in what concentration. The second step is a more extensive screening, aimed at precisely characterising the toxicity of the effluent, the extent of biodegradation that can be achieved, as well as at establishing whether a potential for adaptation of the biomass exists upon exposure. If the sample passes the screening stage, the same serum bottle method will be used to conduct a series of batch co-digestion experiments aimed at evaluating a convenient volumetric ratio between the test material and the readily biodegradable substrate. Finally, a laboratory-scale codigestion trial could simulate the full-scale process, thus enabling the selection of appropriate operating conditions for the start-up of the full-scale implementation. This the protocol has been used to assess the amenability to be anaerobically (co)digested of four industrial effluents, i.e. size and distillery effluents which are classified as high strength and scour and synthetic dye effluents classified as toxic. From the biodegradability and toxicity assays the following conclusions were drawn. The size and distillery effluent were found to be ii degradable at 32 g COD/ℓ and 16 g COD /ℓ concentrations respectively. Concentrations higher than these stipulated above were found inhibitory. Scour effluent was found to be recalcitrant at all concentration tested and synthetic dye was 100 % degradable at 0.12 g COD/ℓ and lower and highly inhibitory at concentration higher than 1.1 g COD/ℓ. Co-digestion experiment using serum bottle AAT method were undertaken between effluents i.e. size + distillery, size + scour, distillery + synthetic dye in an attempt to verify whether the digestion performance benefits from simultaneous presence of the two substrates. The volumetric ratios between the effluents were 1:1, 1:2, 2:1. The presence of two mixtures in the case of size and distillery had better methane production compared to individual substrate i.e. size or distillery separate. The mixture with volumetric flow rate ratio of 2:1 (size: distillery) was preferable in terms of process performance as it had highest COD removal compared to the other mixtures /ratios and individual substrates. The mixture of size and scour (2:1) had highest degradation percentage compared to other ratios but not high enough to qualify as degradable (less than 50 %). The mixture of distillery and synthetic dye had the same pattern with ratio of 2:1 giving the best COD conversion. The pattern than can be drawn from the degradability of mixtures is: the degradability of mixtures increase with the increasing amount of the most biodegradable compound/effluent in the mixture. Serum bottle results provided the detailed information regarding the safe operating parameters which should be used during the starting point for the larger scale investigation i.e. lab-scale investigations. The lab scale investigations were conducted primarily to validate screening and monitor how the digestion progresses and also to provide data for future project i.e. pilot plant investigation. Other effluents i.e. scour and synthetic dye and their co-digestion mixture were excluded from the lab-scale investigations since they were found to be non- biodegradable i.e. their COD conversion was less the 50 % in the screening protocol. Due to time constrains and other technical difficulties in the laboratory, the co-digestion of size and distillery mixture trials we not conducted on the laboratory scale. Laboratory-scale digestion trials showed that the best organic loading rate for distillery effluent in terms of reactor performance and stability was 1.0g COD/ℓ with efficiency of about 45 %, and for size was 2.0g COD/ℓ with an efficiency of 40 %. The efficiencies obtained in both effluents trials could be greatly improved by acclimation; however these results showed that the digestion of these effluents on the bigger scale is possible. / Water Research Commission

Thesis (MSc)--University of Stellenbosch, 2005. / ENGLISH ABSTRACT: Biological removal of sulphate from wastewater can be achieved by using a gas mixture consisting of 80% hydrogen and 20% carbon dioxide as energy and carbon sources. A novel reactor, including a venturi device for optimal hydrogen gas-liquid contact, and geotextile for immobilisation of the sulphate reducing bacterial community, was introduced. Efficient, relatively stable sulphate removal was obtained when the reactor was operated in continuous mode. The maximum sulphate removal rate obtained when the reactor was 8% packed with geotextile, was 1 g S04/(L.d) and 4 g S04/(L.d) when the reactor was 80% packed with geotextile. Kinetic batch studies showed that the highest sulphate removal rates were obtained at 29.5 °C; a pH of 7.5; initial sulphate concentration of 4000 mg/L; initial alkalinity of 1600 mg/L; cobalt concentration of 3 mg/L and when excess hydrogen gas was fed compared to what is stoichiometrically required (900 ml/min). Nickel addition showed inhibition at increased concentrations (>3 mg/L). The biofilm structure was observed on the geotextile with electron microscopy, while the viability of the biofilm was indicated with fluorescence microscopy. These observations indicated the suitability of the geotextile as a support material for biofilm formation in the sulphate reducing system. The stability of the sulphate reducing community was analysed, using the T-RFLP protocol. It was shown that the composition of the community changed after a period of 3 months, when the reactor was subjected to environmental changes. The reactor was also observed to be more efficient in terms of sulphate removal after the environmental changes, of which the temperature change from an average of 39 to 29.5 °C was the most prominent. Subsequently, it was speculated that the population shift was in favour of a more efficient system for sulphate removal. A dynamic, viable, mesophilic sulphate reducing community was therefore observed on the geotextile support, responsible for successful sulphate removal in a novel venturi-reactor. Defining optimal operating conditions, and a knowledge of biofilm structure and composition may contribute to the successful implementation of the biological sulphate removal component of the integrated chemical-biological process for the treatment of industrial wastewater, when hydrogen and carbon dioxide are supplied as the energy and carbon sources, respectively. / AFRIKAANSE OPSOMMING: Ongewenste industriële afval-water kan biologies behandel word deur 'n gasmengsel van 80% waterstof en 20% koolstofdioksied te gebruik vir sulfaat verwydering. 'n Reaktor wat 'n venturi apparaat bevat vir optimale waterstofgas-vloeistof kontak, asook geotekstiel vir die immobilisasie van die bakteriële sulfaatverwyderende gemeenskap, is bekend gestel. Effektiewe, relatief stabiele sulfaatverwydering is waargeneem sodra die reaktor op 'n kontinue basis gevoer is. Die optimale sulfaat verwyderingstempo wat bereik is as die reaktor 8% met geotekstiel gevul was, was 1 g S04/(L.d) en 4 g S04/(L.d) wanneer die reaktor 80% met geotekstiel gevul was. Kinetiese groepstudies het getoon dat die beste sulfaatverwydering bereik is by 'n gemiddelde temperatuur van 29.5 °C; pH van 7.5; aanvanklike sulfaatkonsentrasie van 4000 mg/L; aanvanklike sulfied konsentrasie van 268 mg/L; aanvanklike alkaliniteit van 1600 mg/L; kobalt konsentrasie van 3 mg/L, asook wanneer 'n oormaat waterstofgas gevoer is (900 ml/min), in vergelyking met wat stoichiometries benodig word. 'n Verhoogde byvoeging van nikkel by die voerwater (3 mg/L), het tekens van inhibisie getoon. Die biofilm struktuur is waargeneem op die geotekstiel met behulp van 'n elektronrnikroskoop, terwyl die lewensvatbaarheid van die biofilm aangedui is met behulp van fluoressensie mikroskopie. Hiermee is die bruikbaarheid van geotekstiel as 'n ondersteunings-matriks bevestig. Die stabiliteit van die sulfaatverwyderende gemeenskap is ondersoek deur die T-RFLP protokol te gebruik. Hiermee is aangedui dat die samestelling van die gemeenskap verander het na die 3 maande toets periode, toe die reaktor onderhewig was aan omgewings veranderinge. Die reaktor het ook 'n verbetering in sy sulfaatverwyderings vermoë getoon na hierdie tydperk van omgewingsveranderinge, waarvan 'n temperatuur verandering vanaf 'n gemiddeld van 39 na 29.5 °C die prominentste was. Dit is dus gespekuleer dat die populasie verskuiwing ten gunste was van 'n beter sisteem vir sulfaatverwydering. 'n Dinamiese, lewensvatbare, mesofiliese sulfaatreduserende gemeenskap, verantwoordelik vir die sulfaatverwydering in die venturi-reaktor, is dus waargeneem op die geotekstiel as 'n ondersteuningsmatriks. Met hierdie evaluasie kan die insig wat verkry is in die reaktor samestelling en die optimale kondisies vir die reaktor werking, bydra tot die suksesvolle implementasie van die biologiese komponent, in die geïntegreerde chemies-biologiese proses vir die behandeling van industriële afval water, wanneer 80% waterstof en 20% koolstofdioksied gas as energie en koolstofbron respektiewelik, gebruik word.

Pinch Analysis, in the broadest sense, is concerned with the optimal use of resources (material or energy) in a multi-process system. Pinch Analysis based techniques have emerged for water systems over the past decade. A major assumption that has been made in applying these techniques is that a process system can be segregated into a set of process streams and a set of water streams. With this distinction in place, only the water streams are considered in the Pinch Analysis with the process streams represented implicitly. This approach has obvious limitations in situations where a clear distinction between process streams and water streams cannot be made. The chlor-alkali process is an example of a system in which the clear distinction between process streams and water streams cannot be made. Water is intrinsically involved in the process, serving as a carrier medium for raw materials and eventually becoming part of the products produced by the complex. Hydrochloric acid and caustic soda are reagents which are both used within and produced by the complex. These reagents are required by the process at a range of concentrations and the concentrated reagent is diluted to the required concentrations using demineralised water. Within the chlor-alkali complex, a number of effluents containing the reagent species are available and are typically sent to drain. It is conceivable that these effluents might be recovered and used for dilution purposes instead of demineralised water. This would bring about a reduction in the amount of water and concentrated reagent used and the amount of effluent produced by the complex. Given the economic value of these reagents relative to water, their recovery, if feasible, is likely to dominate the optimal water-use and effluent generation strategy. Current Water Pinch Analysis theory relies on the distinction being made between process streams and water streams and does not consider the recovery of reagents or the presence of desirable species within the system. In addition, the assumption is made that species are non-reactive; reactive species such as hydrogen chloride and sodium hydroxide, fall outside the scope of the current theory. The objectives of this study have included the development of an approach which is able to address these limitations of the existing theory. This approach, termed Combined Water and Materials Pinch Analysis seeks to identify optimal use strategies for raw materials and reagents, in addition to water-use and effluent generation. The approach combines mathematical programming with conceptual insights from Water Pinch Analysis. The approach is based on the optimisation of a superstructure which represents the set of all possible flow configurations for water, reagents and raw materials between the various operations within the process system; this problem is solved as a nonlinear programming (NLP) problem using standard optimisation tools. The application of the developed approach to the Sasol Polymers chlor-alkali complex at Umbogintwini, south of Durban, has been a further objective of this study. Given the variety of process operations present within the complex, which differ both in terms of their physical structure and function, individual process models for these operations were required. These models were described in terms of four basic functional elements, namely, mixing, flow separation, component separation and reaction, and incorporated into the superstructure. Given the complexity of the problem, the process system was divided into three subsystems which were optimised in isolation from each other. These results were subsequently integrated to reflect the performance of the subsystems in combination with each other. The results showed a potential reduction of 14% in water-use and 42% in effluent production by the complex, relative to the existing operating configuration. Amongst other savings in material use, the results indicated a 0.2% reduction in the use of salt, a 1.6% reduction caustic soda use and an 8.3% reduction in the use of hydrochloric acid. Economically, the potential saving identified was R 945 727 per annum, based on operating costs in the year 2000. The final objective of this study was the interpretation of the pinch as it relates to the Combined Water and Materials Pinch Analysis problem. A general definition of the pinch was proposed; according to this definition, the pinch corresponds to that constraint or set of constraints which limits the performance of the system, that is, prevents it from further improvement. For the Combined Water and Materials Pinch Analysis problem, this performance is measured in terms of the operating cost. This definition is thus a departure from its usual thermodynamic interpretation of the pinch; in addition, the pinch is defined in terms of a constraint or a set of constraints instead of a point. These constraints are identified by an analysis of the marginal values provided by the optimisation algorithm. Marginal values are also used as a means of identifying process interventions which may be effected such that the performance of the system may be improved further. / Thesis (M.Sc.Eng.)-University of Natal, 1989.

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.

Submitted in partial fulfillment of academic requirements for the degree of Masters of Technology: Department of Chemical Engineering, Durban University of Technology, 2009. / The primary objective of this project was to establish a screening protocol which could be used to access high strength/toxic effluent for toxicity and degradability prior to being disposed in wastewater treatment works. The serum bottle method (materials and method section) is simple, makes use of small glass vials (125 mℓ-volume were used in this research) which do not require any stirring nor feeding device or other engineered tool: a serum bottle is sealed immediately after all components are poured inside and thereafter conducted in a batch mode and occasionally shaken to ensure adequate homogenisation of the components. The only variables which are regularly measured are the volume of biogas produced and gas composition. The two assays, originally developed by Owen et al. (1979) to address the toxicity and the biodegradability have been combined in a single test called AAT, Anaerobic Activity Test, which enables one to assess simultaneously the inhibitory effect on the methanogenic biomass and the biodegradability of the test material as well as the ability of the biomass to adapt to the test material and therefore to overcome the initial inhibition. The screening protocol is illustrated in Annexure A. The protocol consists of a sequence of assays which employ the serum bottle methodology. A first step of the procedure is aimed at rapidly estimating whether the effluent is potentially toxic to the methanogenic biomass and in what concentration. The second step is a more extensive screening, aimed at precisely characterising the toxicity of the effluent, the extent of biodegradation that can be achieved, as well as at establishing whether a potential for adaptation of the biomass exists upon exposure. If the sample passes the screening stage, the same serum bottle method will be used to conduct a series of batch co-digestion experiments aimed at evaluating a convenient volumetric ratio between the test material and the readily biodegradable substrate. Finally, a laboratory-scale codigestion trial could simulate the full-scale process, thus enabling the selection of appropriate operating conditions for the start-up of the full-scale implementation. This the protocol has been used to assess the amenability to be anaerobically (co)digested of four industrial effluents, i.e. size and distillery effluents which are classified as high strength and scour and synthetic dye effluents classified as toxic. From the biodegradability and toxicity assays the following conclusions were drawn. The size and distillery effluent were found to be ii degradable at 32 g COD/ℓ and 16 g COD /ℓ concentrations respectively. Concentrations higher than these stipulated above were found inhibitory. Scour effluent was found to be recalcitrant at all concentration tested and synthetic dye was 100 % degradable at 0.12 g COD/ℓ and lower and highly inhibitory at concentration higher than 1.1 g COD/ℓ. Co-digestion experiment using serum bottle AAT method were undertaken between effluents i.e. size + distillery, size + scour, distillery + synthetic dye in an attempt to verify whether the digestion performance benefits from simultaneous presence of the two substrates. The volumetric ratios between the effluents were 1:1, 1:2, 2:1. The presence of two mixtures in the case of size and distillery had better methane production compared to individual substrate i.e. size or distillery separate. The mixture with volumetric flow rate ratio of 2:1 (size: distillery) was preferable in terms of process performance as it had highest COD removal compared to the other mixtures /ratios and individual substrates. The mixture of size and scour (2:1) had highest degradation percentage compared to other ratios but not high enough to qualify as degradable (less than 50 %). The mixture of distillery and synthetic dye had the same pattern with ratio of 2:1 giving the best COD conversion. The pattern than can be drawn from the degradability of mixtures is: the degradability of mixtures increase with the increasing amount of the most biodegradable compound/effluent in the mixture. Serum bottle results provided the detailed information regarding the safe operating parameters which should be used during the starting point for the larger scale investigation i.e. lab-scale investigations. The lab scale investigations were conducted primarily to validate screening and monitor how the digestion progresses and also to provide data for future project i.e. pilot plant investigation. Other effluents i.e. scour and synthetic dye and their co-digestion mixture were excluded from the lab-scale investigations since they were found to be non- biodegradable i.e. their COD conversion was less the 50 % in the screening protocol. Due to time constrains and other technical difficulties in the laboratory, the co-digestion of size and distillery mixture trials we not conducted on the laboratory scale. Laboratory-scale digestion trials showed that the best organic loading rate for distillery effluent in terms of reactor performance and stability was 1.0g COD/ℓ with efficiency of about 45 %, and for size was 2.0g COD/ℓ with an efficiency of 40 %. The efficiencies obtained in both effluents trials could be greatly improved by acclimation; however these results showed that the digestion of these effluents on the bigger scale is possible.

In an attempt to overcome the high maintenance and costs associated with traditional physico-chemical methods, much work is being done on the application of enzymes for the recovery of valuable metals from solutions and industrial effluents. One of the most widely studied enzymatic metal recovery systems uses hydrogenase enzymes, particularly from sulphate reducing bacteria (SRB). While it is known that hydrogenases from SRB mediate the reductive precipitation of metals, the mechanism of enzymatic reduction, however, is not yet fully understood. The main aim of the present study was to investigate the role of a hydrogenase enzyme in the removal of rhodium from both aqueous solution and industrial effluent. A quantitative analysis of the rate of removal of rhodium(III) by a resting SRB consortium under different initial rhodium and biomass concentrations, pH, temperature, presence and absence of SRB cells and electron donor, was studied. Rhodium speciation was found to be the main factor controlling the rate of removal of rhodium from solution. SRB cells were found to have a higher affinity for anionic rhodium species, as compared to both cationic and neutral species, which become abundant when speciation equilibrium was reached. Consequently, a pH-dependant rate of rhodium removal from solution was observed. The maximum SRB uptake capacity for rhodium was found to be 66 mg rhodium per g of resting SRB biomass. Electron microscopy studies revealed a time-dependant localization and distribution of rhodium precipitates, initially intracellularly and then extracellularly, suggesting the involvement of an enzymatic reductive precipitation process. A hydrogenase enzyme capable of reducing rhodium(III) from solution was isolated and purified by PEG, DEAE-Sephacel anion exchanger and Sephadex G200 gel exclusion. A distinct protein band with a molecular weight of 62kDa was obtained when the hydrogenase containing fractions were subjected to a 10% SDS-PAGE. Characterization studies indicated that the purified hydrogenase had an optimum pH and temperature of 8 and 40°C, respectively. A maximum of 88% of the initial rhodium in solution was removed when the purified hydrogenase was incubated under hydrogen. Due to the low pH of the industrial effluent (1.31), the enzymatic reduction of rhodium by the purified hydrogenase was greatly retarded. It was apparent that industrial effluent pretreatment was necessary before the application an enzymatic treatment method. In the present study, however, it has been established that SRB are good candidates for the enzymatic recovery of rhodium from both solution and effluent.

The continuing industrial development has led to a corresponding increase in the amount of waste water generation leading to a consequential decline in levels and quality of the natural water in the ecosystem. Textile industries consume over 7 x 10[superscript 5] tons of dyes annually and use up to 1 litre of water per kg of dye processed and are third largest polluters in the world, the problem being aggravated by the inefficiencies of the dye houses. An abundance of physio-chemical methods are in use world wide, however, there is increasing concern as to their impact in effectively treating textile effluents as they introduce secondary pollutants during the ‘remediation’ process which are quite costly to run, maintain and clean up. Research on biological treatment has offered simple and cost effective ways of bioremediating textile effluents. While aerobic treatment of textile dyes and their effluents has been reported, its major draw back is commercial up-scaling and as such anaerobic systems have been investigated and shown to degrade azo dyes, which form the bulk of the dyes used world wide. However, the mechanisms involved in the bioremediation of these dyes are poorly understood. The aims of this study were to identify and investigate the role of enzymes produced by sulphate reducing bacteria (SRB) in bioremediating textile dye and their effluents. Sulphate reducing bacteria were used in this study because they are tolerant to harsh environmental conditions and inhibit the proliferance of pathogenic micro-organisms. The appearance of clear zones in agar plates containing azo dye concentrations ranging from 10 – 100 mgl[superscript -1] showed the ability of SRB to decolourize dyes under anaerobic conditions. Assays of enzymes previously reported to decolourise azo dyes were not successful, but led to the identification of hydrogenase enzyme being produced by SRB. The enzyme was found to be localised in the membrane and cytoplasm. A surface response method was used to optimize the extraction of the enzyme from the bacterial cells resulting in approximately 3 fold increase in hydrogenase activity. Maximum hydrogenase activity was found to occur after six days in the absence of dyes but was found to occur after one day in the presence of azo dyes. A decline in hydrogenase activity thereafter, suggested inhibition of enzymatic activity by the putative aromatic amines produced after azo cleavage. Purification of the hydrogenase by freeze drying, poly ethylene glycol, and Sephacryl – 200 size exclusion- ion exchange chromatography revealed the enzyme to have a molecular weight of 38.5 kDa when analyzed by a 12 % SDS-PAGE. Characterisation of the enzyme revealed optimal activity at a pH of 7.5 and temperature of 40 °C while it exhibited a poor thermal stability with a half-life of 32 minutes. The kinetic parameters V[subscript max] and K[subscript m] were 21.18 U ml[superscript -1} and 4.57 mM respectively. Application of the cell free extract on commercial dyes was not successful, and only whole SRB cells resulted in decolourisation of the dyes. Consequently trials on the industrial dyes and effluents were carried out with whole cells. Decolourisation rates of up to 96 % were achieved for the commercial dyes and up to 93 % for the industrial dyes over a period of 10 days.

Thesis (MSc Food Sc)--Stellenbosch University, 2000. / ENGLISH ABSTRACT: The distillery industry produces large volumes of waste water with a high organic content throughout the year. These effluents must be treated in some manner before being discharged or recycled in the factory. Several treatment options are in use presently, but they all have disadvantages of some nature, such as long retention times, bad odours or the need for large areas of land. Considerable interest has been shown in the application of anaerobic digestion, especially the UASB design (upflow anaerobic sludge blanket), to treat this high strength waste water. Thus, the aim of this study was to investigate the efficiency of an upflow anaerobic sludge blanket (UASB) bioreactor using full-strength distillery effluent. The activity of the bacteria in the biogranules was also evaluated by developing an easy and reliable activity method to estimate the general biogas and methanogenic activity and to calibrate this method using different anaerobic granules from different sources. The influence of high strength distillery effluent on the anaerobic digestion process was investigated using a mesophilic lab-scale UASB bioreactor. During the experimental study, the organic loading rate (OLR) was gradually increased from 2.01 to 30.00 kgCOD.m-3.d-1, and simultaneously, the substrate pH was gradually lowered from 7.0 to 4.7. It was found that at an OLR of 30.00 kgCOD. m-3.d-1,the pH, alkalinity and biogas production stabilised to average values of 7.8, 6 000 mg.l-1 and 18.5 I.d-1 respectively. An average COD removal> 90% was found indicating excellent bioreactor stability. The low substrate pH holds considerable implications in terms of operational costs, as neutralisation of the biorector substrate is no longer necessary. The accumulation of fine solids present in the distillery substrate was found at the higher OLR's and resulted in the granular bed increasing with subsequent biomass washout and a lowering in efficiency parameters. However, a possible pre-treatment filtration of these fine solids would eliminate this problem. The success of the upflow anaerobic sludge bed (UASB) process is mainly due to the capability of retaining the active biomass in the reactor. Over the years, several methods have been developed to characterise and quantify sludge activity but each has advantages and disadvantages. There is thus an increasing need for a rapid method to evaluate the activity of the granular biomass. The activity method of Owen et al. (1979) as adapted by Lamb (1995), was thus evaluated in terms of efficiency and applicability in determining the activity of granular samples. The method was found to be inaccurate as well as time consuming and it was thus modified. Results obtained with the modified assay method were found to be more accurate and the impact of the different test substrates (glucose, lactate, acetate and formate) on activity, was more evident. The activity of seven different anaerobic granules, was subsequently evaluated. Biogas (Ss) and methanogenic (SM) activity was not measured in volume of gas produced per unit COD converted or volatile suspended solids (VSS), but as tempo of gas production (ml.h-1) in a standardised basic growth medium. The activity data obtained were also displayed as bar charts and "calibration scales". This illustrative depiction of activity data gave valuable information about population dynamics as well as possible substrate inhibition. The "calibration scales" can also be used to group the general biogas (Ss) and methanogenic activities (SM) of any new biogranule relative to active (O-type) and inactive (W-type) anaerobic granules, providing that the same method of activity testing is used. The "calibration scales" can thus be used to give a fast indication of how the activity value of one sample relates to the activity values of other granules, even when using different test substrates. / AFRIKAANSE OPSOMMING: Die stokery industrie produseer groot hoeveelhede afvalwater, wat hoë ladings van organiese materiaal gedurede die hele jaar bevat. Hierdie afvalwater moet op een of ander manier behandel word voordat dit gestort of vir hergebruik aangewend kan word. Daar is tans verskeie behandelingsmetodes wat gebruik kan word, maar elk het sy eie tekortkominge soos bv. lang retensie tye, onaangename reuke of die behoefte aan groot stukke oop grond. Groot belangstelling is getoon vir die gebruik van anaerobiese vertering, en meer spesifiek die "uflow anaerobic sludge blanket" UASB bioreaktor vir die behandeling van stokery uitvloeisels. Die doel van die studie was dus om die algehele effektiwiteit van 'n UASB bioreaktor, wat onverdunde stokery uitvloeisel behandel, te evalueer. Die methanogene- en algehele aktiwiteit van die bakterië in die biogranules was ook ge-evalueer deurdat 'n maklike en betroubare aktiwiteitsmetode omtwikkel is, waarna hierdie metode ook toegepas was op 'n reeks van verskillende tipe biogranules. Die invloed van volsterkte stokery uitvloeisel op die anaerobiese verteringsprosesse was ondersoek met die gebruik van 'n mesofiele laboratoriumskaal UASB bioreaktor. Gedurende die eksperimentele studie, was die organiese ladingstempo (OLT) verhoog van 2.01 na 30.00 kgCSB.m-3.d-1 (CSB = chemiese suurstof behoefte) met die gelyktydige verlaging in die pH van die bioreaktorsubstraat van 7.0 na 4.7. Dit was vasgestel dat met 'n OLT van 30.00 kgCSB.m-3.d-1, die pH, alkaliniteit en biogas geproduseer, gestabiliseer het na gemiddelde waardes van 7.8, 6000 mg.-1 en 18.5l.d-1 , respektiewelik, sowel as 'n gemiddelde CSB verwydering van> 90%. Al hierdie waardes dui uitstekende bioreaktor stabiliteit aan. Die lae bioreaktorsubstraat pH kan van groot waarde wees vir die industrie, aangesien neutralisering van die uitvloeisel nie meer nodig is nie en kan sodoende die operasionele koste van die proses verlaag. Die konsentrering van fyn opgeloste soliedes in die bioreaktor by hoë OLT's, kan egter problematies raak, aangesien dit die granule-bed kan vergroot en veroorsaak dat van die biomassa uitspoel en kan verlore gaan. Die verlies van aktiewe biomassa kan die effektiwiteitsparameters negatief beinvloed, maar die plasing van 'n filterings stap voor die verterings stap, behoort hierdie probleem op te los. The sukses van die UASB-stelsel rus op die versekering dat die aktiewe biomassa in die reaktor behoue bly. Oor die jare was daar 'n verskeidenheid van aktiwiteitstoetsings-metodes ontwikkel, elk met sy eie nadele. Daar bestaan dus nog steeds 'n groot behoefte vir die daarstelling van 'n aktiwiteitstoetsings-metode wat vinnig en maklik is om uittevoer. Die aktiwiteitstoetsings-metode van Owen et al. (1979) wat deur Lamb (1995) aangepas is, was in terme van sy effektiwiteit en toepaslikheid ten opsigte van die gebruik daarvan vir aktiwiteitstoetsing vir biogranules, ge-evalueer. Dit is bevind dat die metode onakkuraat sowel as tydsrowend was en gevolglik dus aangepas. Die aangepaste metode het meer akkurate resultate gelewer en die impak van die verskillende toetssubstrate (glukose, laktaat, asetaat en formaat) op die granules het ook meer duidelik na vore gekom. Gevolglik was die aktiwiteit van sewe verskillende anaerobiese biogranules ondersoek. Die eenheid waarin atiwiteitsresultate aangegee is, was nie in volume gas geproduseer per eenheid CSB verwyder of per hoeveelheid gesuspendeerde vlugtige vetsure in die biomassa nie, maar as tempo van biogas (S8)- of metaan (SM)produksie (ml.h-1). Die data wat op hierdie wyse bekom was, is gebruik om staafdiagramme sowel as "kalibrasie skale" daar te stel. Hierdie illustrerende wyse om aktiwiteitsdata uit te beeld verskaf waardevolle informasie ten opsigte van die interaksies tussen die verskillende populasies in die granule en kan ook die aanwesigheid van moontlike substraat inhibisie aandui. Die "Kalibrasie skale" kan ook gebruik word om die algehele (SB) en methanogene (SM)aktiwiteite van einge nuwe biogranule vinnig te klassifiseer ten op sigte van 'n aktiewe (O-tipe) en 'n minder aktiewe (W-tipe) anaerobiese granules, mits dieselfde metode gebruik word om die aktiwiteits data te bekom.

CAPES / As indústrias de celulose são caracterizadas pelo alto consumo de água em seus processos produtivos, gerando consequentemente grandes volumes de efluentes líquidos que apresentam na maior parte de sua composição compostos lignínicos, matéria orgânica, cor e toxicidade. O efluente de celulose, quando não tratado ou tratado de forma indevida, pode comprometer a qualidade da água dos corpos receptores, por conter substâncias tóxicas à comunidade aquática. O objetivo do trabalho foi avaliar a eficiência de tratamento de efluente de celulose e papel por reator MBBR, através da remoção de matéria orgânica (DQO e DBO5), compostos fenólicos, cor, compostos lignínicos e aromáticos. Para isso um reator MBBR em escala de bancada foi operado por 180 dias com diferentes velocidades de carga orgânica 0,2; 0,4; 1,2; 4,0; 9,0 kgDQO/L.d. Neste foi quantificada também a biomassa aderida e em suspensão na massa liquida. O efluente analisado apresentou 48 e 94% de remoção de DQO e DBO5 respectivamente, também houve remoção de compostos fenólicos e cor de 24 e 12% para VCO de 0,4 kgDQO/L.d, para mesma VCO a remoção de compostos lignínicos e aromáticos foi de 16 e 8,5 % respectivamente. / The pulp mills are characterized by high water consumption their production processes, thereby generating large volumes of effluents that present in most of composition lignínicos compounds, organic matter, color and toxicity. The wastewater from pulp, if left untreated or treated improperly, can compromise the water quality of receiving waters, which contain substances toxic to the aquatic community. The objective of this study was to evaluate the efficiency of wastewater treatment of pulp and paper by MBBR reactor, through the removal of organic matter (COD and BOD5), phenolic compounds, color, and aromatic compounds lignínicos. To this reactor MBBR bench scale was operated for 180 days with different speeds organic load 0.2, 0.4, 1.2, 4.0, 9.0 kgCOD / Ld This was also quantified and attached biomass in suspension in the liquid mass. The wastewater analysis showed 48 and 94% removal of COD and BOD5 respectively, were also removing phenolic compounds and color of 24 and 12% for 0.4 kgDQO VCO / Ld VCO same for removal of aromatic compounds and was lignínicos 16 and 8.5% respectively.