The removal of multiple substrates in a defined mixed culture process was investigated in the treatment of brewery wastewater. The study was conducted using both batch and a semi-continuous reactor system called self-cycling fermentation. Batch experiments were conducted using a synthetic brewery wastewater containing glucose, ethanol and maltose. Activated sludge from a municipal wastewater treatment plant was acclimatized in the synthetic brewery wastewater. The microbes capable of degrading this wastewater were analyzed by a combination of microscopy, spread plating, and Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis (PCR-DGGE) and identified as Acinetobacter sp., Enterobacter sp. and Candida sp. From the pure culture batch experiments, it was determined that Enterobacter could degrade glucose and maltose but no ethanol, while Acinetobacter and Candida could degrade all three carbon sources. In mixed culture batch experiments, Enterobacter was dominant in degrading the sugar concentrations to levels permissible for Acinetobacter to degrade ethanol. PCR-DGGE was found to be effective in identifying the dominant species but selective carbon source plating was required to determine viability and track the population dynamics. Kinetic experiments were carried out in a semi-continuous, self-cycling fermentation process using the defined mixed culture in media containing glucose and various initial concentrations of ethanol and maltose. The overall rate of substrate removal was attributable to both the suspended culture and the biofilm formed during the process. A rate expression was developed for this system for the range of substrate concentrations tested. The data indicated that substrate removal by the suspended culture was a function of only the biomass concentration. However, substrate removal by the biofilm was found to be limited to the surface cells and determined to be a function of substrate concentration only.

There is potential for the anaerobic baffled reactor (ABR) to be implemented on-site for pre-treatment of coloured wastewaters. The implementation of waste minimisation and cleaner production strategies in industry will result in the production of smaller volumes of concentrated wastewaters. With implementation of the ABR, the concentrated waste stream could be pre-treated, with an acclimated biomass, which should facilitate sufficient degradation such that the effluent could be discharged to sewer for further treatment. The ABR is a high-rate compartmentalised anaerobic bioreactor, the design of which promotes the spatial separation of microorganisms. The use of molecular teclmiques to characterise the microbial populations and the dynamics of these populations with time and/or changing operating conditions will add to the current understanding of the process, which is based on the biochemical pathways and chemical analyses. This knowledge will allow for optimisation of the design of the ABR. The hypothesis of the horizontal separation of acidogenesis and methanogenesis through the ABR was proven. Changes in the HRT affected the operation of the reactor, however, recovery from these upsets was almost immediate and operation of the reactor was stable. Two synthetic dye waste streams, one food dye (tartrazine) and one textile dye (Cl Reactive Red 141), and a real industrial dye wastewater, were treated in separate laboratory-scale ABRs. These investigations showed that successful treatment of a highly coloured wastewater is possible in the ABR. The design of the ABR facilitates efficient treatment of concentrated dye wastewaters by protecting the sensitive methanogens from the inhibitory dye molecules and promoting efficient colour and COD reduction. The molecular-based method, fluorescent in situ hybridisation, allowed the direct identification and enumeration of microbial populations active in the ABR. In all of the reported investigations, there was a definite shift in the microbial populations through the ABR, with a predominance of eubacteria in the first compartments (acidogenesis) and archaea (methanogenesis) in the later compartments. The number of compartments involved in each depended on the strength of the substrate (organic loading rate - OLR). A combination of FISH probing, and the analysis of 98 archaeal l6S rDNA clone inserts provided useful descriptions of the methanogens actively involved within each compartment. These showed a predominance of the Methanosaeta spp., particularly in the last compartments of the reactor. Methanogens present in the first four compartments consisted of species of Methanobacterium and Methanospirillum, a relatively unstudied methanogen Methanomethylovorans hollandica, and an unidentified short filamentous species. / Thesis (Ph.D.)-University of Natal, Durban, 2002.

Self-cycling fermentation (SCF) control was applied to a semi-continuous, aerobic, fixed-film reactor for the treatment of simulated and real brewery wastewater. The simulated wastewater was treated at 25°C. After approximately 3 hrs, 86% to 96% of the soluble BOD (SBOD) was removed, producing an effluent with a SBOD between 169 mg/L and 412 mg/L. The treatment of real brewery wastewater was undertaken at 25°C and 35°C, and was operated under ammonia-nitrogen deficient condition. 83% to 92% of the total BOD (TBOD) was removed after 3 hrs at 25°C, and after 1.5 to 2 hrs at 35°C. The treated effluent produced had a TBOD between 120 and 438 mg/L. The suspended solids in the treated effluent contributed between 63% and 71% of the TBOD. Hence, better treatment efficiency would have been possible if an efficient clarifier was installed, or the suspended solids were removed from the brewery wastewater prior to treatment.

Inadequate sanitation is one of the leading causes of water pollution and consequently illness in many underdeveloped countries, including South Africa and, specifically, the Eastern Cape Province, where cholera has become endemic. As modern wastewater treatment processes are often energy intensive and expensive, they are not suitable for use in these areas. There is thus a need to develop more sustainable wastewater treatment technologies for application in smaller communities. The integrated algal ponding system (IAPS) was identified as a possible solution to this wastewater management problem and was investigated for adaptation to local conditions, at the Rhodes University Environmental Experimental Field Station in Grahamstown, South Africa. The system was monitored over a period of nine years, with various configuration adjustments of the high rate algal pond (HRAP) unit operation investigated. Under standard operating conditions, the system was able to achieve levels of nutrient and organic removal comparable with conventional wastewater treatment works. The mean nitrate level achieved in the effluent was below the 15mg.l-1 South African discharge standard, however, nitrate removal in the IAPS was found to be inconsistent. Although the system was unable to sustain chemical oxygen demand (COD) removal to below the 75mg.l-1 South African discharge standard, a removal rate of 87% was recorded, with the residual COD remaining in the form of algal biomass. Previous studies in the Eastern Cape Province have shown that few small wastewater treatment works produce effluent that meets the microbial count specification. Therefore, in addition to the collation of IAPS data from the entire nine year monitoring period, this study also investigated the use of the HRAP as an independent unit operation for disinfection of effluent from small sewage plants. It was demonstrated that the independent high rate algal pond (IHRAP) as a free standing unit operation could consistently produce water with Escherichia coli counts of 0cfu.100ml-1. The observed effect was related to a number of possible conditions prevailing in the system, including elevated pH, sunlight and dissolved oxygen. It was also found that the IHRAP greatly enhanced the nutrient removal capabilities of the conventional IAPS, making it possible to reliably and consistently maintain phosphate and ammonium levels in the final effluent to below 5mg.l-1 and 2mg.l-1 respectively (South African discharge standards are 10mg.l-1 and 3mg.l-1 in each case). The quality of the final effluent produced by the optimisation of the IAPS would allow it to be used for irrigation, thereby providing an alternative water source in water stressed areas. The system also proved to be exceptionally robust and data collected during periods of intensive and low management regimes were broadly comparable. Results of the 9 year study have demonstrated reliable performance of the IAPS and its use an appropriate, sustainable wastewater treatment option for small communities.

The research reported here investigated the use of a Baffle Reactor in order to study aspects of the biological sulfur cycle, where a floating sulfur biofilm formation occurs and where complex organic compounds provide electron donor sources. The development of a laboratory-scale Baffle Reactor model system satisfied the requirements for sulfate reducing bacterial biomass growth and sulfur biofilm formation. Since relatively little is known about the microbial ecology of floating sulfur biofilm systems, this study was undertaken to describe the sulfate reducing sludge population of the system together with its performance. A combination of culture- and molecular-based techniques were applied in this study in order to investigate the microbial ecology of the sulfate-reducing bacteria component of the system. These techniques enabled the identification and the analysis of the distribution of different sulfate reducing bacterial strains found within the sludge bioreactors. Strains isolated from the sludge were characterised based on culture appearance, gram staining and scanning electron microscopy morphology. Molecular methods based on the PCR-amplified 16S rRNA including denaturing gradient gel electrophoresis were employed in order to characterise sulfate-reducing bacteria within the reactors. Three novel Gram negative sulfate-reducing bacteria strains were isolated from the sludge population. Strains isolated were tentatively named Desulfomonas rhodensis, Desulfomonas makanaiensis, and Clostridium sulforhodensis. Results obtained from the Baffle Reactor showed that three dominant species were isolated from the DNA extracted from the whole bacterial population by peR. Three of these were similar to those mentioned above. The presence of these three novel unidentified species suggest that there are a range of other novel organisms involved in sulfate reduction processes.

Phenolic compounds are a group of organic chemicals present in the wastewaters of many synthetic industrial processes. Due to their extreme toxicity to man and animals, and deleterious impact on the environment, a range of techniques exist for the effective treatment and disposal of these pollutants. Biological degradation using microbial enzymes presents a valuable alternative to conventional wastewater treatment systems. This research was therefore initiated to investigate the polyphenol oxidase enzyme system and the feasibility of its application for effluent treatment and studies in organic solvents. The enzyme system is widely distributed in nature, with Agaricus bisporus (the common mushroom) being the best known producer. Biochemical investigations of the enzyme system were therefore carried out using this extract. A screening programme was initiated to identify microbial polyphenol oxidase producers which could be cultured in liquid media, thereby enabling the production of large quantities of enzyme in fermentation systems. Extensive growth optimization and enzyme induction and optimization studies were carried out on selected cultures. A number of good producers were isolated, namely a bacterial culture designated AECI culture no. 26, Streptomyces antibioticus, Streptomyces glaucescens and a manipulated strain, Streptomyces lividans (pIJ702). Enzyme production by Agaricus bisporus mycelia was optimized in deep-liquid culture; enzyme extracts showed high phenol removal efficiencies. Streptomyces antibioticus, Streptomyces glaucescens, Streptomyces lividans (pIJ702) and AECI culture no. 26 whole cells were also investigated for phenol-removing ability in simulated phenolic effluents. The use of whole cells reduces enzyme inactivation and instability due to the protection of the enzyme system within the cell. All cultures showed improved removal efficiencies in phenolic growth media. These results strongly suggest their use for phenol removal in continuous systems.

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

No description available.

No description available.

M. S.