The microbial ecology of sulphidogenic lignocellulose degradation
- Authors: Clarke, Anna Maria
- Date: 2007
- Subjects: Microbial ecology , Lignocellulose , Sulfides , Lignin , Lignocellulose -- Biodegradation , Mines and mineral resources -- Waste disposal , Acid mine drainage
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4094 , http://hdl.handle.net/10962/d1008181
- Description: Acid mine drainage is a well known environmental pollutant, not only in South Africa, but throughout the world, and the use of microbial processes in the treatment of these wastes has been the subject of investigation over past decades. Lignocellulose packed-bed reactors have been used in passive treatment systems, and, although effective initially, they show early decline in performance while the packing material remains largely un-utilized. Little is known about this phenomenon which remains a severe constraint in the development of efficient passive mine water treatment systems. It has been proposed that the degradation pathways of the complex lignocellulose substrate may be limited in some way in these systems during the manifestation of this effect. This study has addressed the problem using a molecular microbial ecology methodology in an attempt to relate trophic functions of the microbial population to the physico-chemical data of the system. A field-scale lignocellulose packed-bed reactor located at Vryheid Coronation Colliery (Northern Kwa-Zulu Natal province, South Africa) was monitored for six years and the results showed the classic profile of performance decline related to a slowdown in sulphate reduction and alkalinity production. The reactor was decommissioned , comprehensive samples were collected along the depth profile and the microbial populations investigated by means of 16S rRNA gene methodology. The population was found to include cellulolytic Clostridia spp., CytophagaIFlavobacterlBacteroidetes, Sphingomonadaceae and as yet uncultured microorganisms related to microbiota identified in the rumen and termite gut. These are all known to be involved as primary fermenters of cellulose. Oesulphosporosinus was present as sulphate reducer. A comparison of substrata sampling and population distribution suggested that spatial and temporal gradients within the system may become established over the course of its operation. Based on these findings, a laboratory-scale reactor was constructed to simulate the performance of the packed-bed reactor under controlled experimental conditions. The laboratory-scale reactor was operated for 273 days and showed comparable performance to that in the field in both biomolecular and physicochemical data. Clearly defined trophic niches were observed. These results suggested that a sequence of events does occur in lignocellulose degradation over time. Based on the spatial and temporal column studies, a descriptive model was proposed to account for these events. It was found that fermentative organisms predominate in the inlet zone of the system using easily extractable compounds from the wood, thus providing feedstock for sulphate reduction occurring in the succeeding compartments. Production of sulphide and alkalinity appears to be involved in the enhancement of lignin degradation and this, in turn, appears to enhance access to the cellulose fraction. However, once the readily extractables are exhausted, the decline in sulphide and alkalinity production leads inexorably to a decline in the overall performance of the system as a sulphate reducing unit operation. These observations led to the proposal that with the addition of a limited amount of a readily available carbon source, such as molasses, in the initial zone of the the reactor, the ongoing generation of sulphide would be sustained and this in turn would sustain the microbial attack on the lignocellulose complex. This proposal was tested in scale-up studies and positive results indicate that the descriptive model may, to some extent, provide an account of events occurring in these systems. The work on sustaining lignocellulose degradation through the maintenance of sulphate reduction in the initial stages of the reactor flow path has led to the development of the Degrading Packed-bed Reactor concept and that, has subsequently been successfully evaluated in the field.
- Full Text:
- Date Issued: 2007
- Authors: Clarke, Anna Maria
- Date: 2007
- Subjects: Microbial ecology , Lignocellulose , Sulfides , Lignin , Lignocellulose -- Biodegradation , Mines and mineral resources -- Waste disposal , Acid mine drainage
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4094 , http://hdl.handle.net/10962/d1008181
- Description: Acid mine drainage is a well known environmental pollutant, not only in South Africa, but throughout the world, and the use of microbial processes in the treatment of these wastes has been the subject of investigation over past decades. Lignocellulose packed-bed reactors have been used in passive treatment systems, and, although effective initially, they show early decline in performance while the packing material remains largely un-utilized. Little is known about this phenomenon which remains a severe constraint in the development of efficient passive mine water treatment systems. It has been proposed that the degradation pathways of the complex lignocellulose substrate may be limited in some way in these systems during the manifestation of this effect. This study has addressed the problem using a molecular microbial ecology methodology in an attempt to relate trophic functions of the microbial population to the physico-chemical data of the system. A field-scale lignocellulose packed-bed reactor located at Vryheid Coronation Colliery (Northern Kwa-Zulu Natal province, South Africa) was monitored for six years and the results showed the classic profile of performance decline related to a slowdown in sulphate reduction and alkalinity production. The reactor was decommissioned , comprehensive samples were collected along the depth profile and the microbial populations investigated by means of 16S rRNA gene methodology. The population was found to include cellulolytic Clostridia spp., CytophagaIFlavobacterlBacteroidetes, Sphingomonadaceae and as yet uncultured microorganisms related to microbiota identified in the rumen and termite gut. These are all known to be involved as primary fermenters of cellulose. Oesulphosporosinus was present as sulphate reducer. A comparison of substrata sampling and population distribution suggested that spatial and temporal gradients within the system may become established over the course of its operation. Based on these findings, a laboratory-scale reactor was constructed to simulate the performance of the packed-bed reactor under controlled experimental conditions. The laboratory-scale reactor was operated for 273 days and showed comparable performance to that in the field in both biomolecular and physicochemical data. Clearly defined trophic niches were observed. These results suggested that a sequence of events does occur in lignocellulose degradation over time. Based on the spatial and temporal column studies, a descriptive model was proposed to account for these events. It was found that fermentative organisms predominate in the inlet zone of the system using easily extractable compounds from the wood, thus providing feedstock for sulphate reduction occurring in the succeeding compartments. Production of sulphide and alkalinity appears to be involved in the enhancement of lignin degradation and this, in turn, appears to enhance access to the cellulose fraction. However, once the readily extractables are exhausted, the decline in sulphide and alkalinity production leads inexorably to a decline in the overall performance of the system as a sulphate reducing unit operation. These observations led to the proposal that with the addition of a limited amount of a readily available carbon source, such as molasses, in the initial zone of the the reactor, the ongoing generation of sulphide would be sustained and this in turn would sustain the microbial attack on the lignocellulose complex. This proposal was tested in scale-up studies and positive results indicate that the descriptive model may, to some extent, provide an account of events occurring in these systems. The work on sustaining lignocellulose degradation through the maintenance of sulphate reduction in the initial stages of the reactor flow path has led to the development of the Degrading Packed-bed Reactor concept and that, has subsequently been successfully evaluated in the field.
- Full Text:
- Date Issued: 2007
Integrated anaerobic/aerobic bioprocess environments and the biodegradation of complex hydrocarbon wastes
- Authors: Ehlers, George A C
- Date: 2004
- Subjects: Hydrocarbons -- Biodegradation Sewage -- Purification -- Anaerobic treatment Water -- Purification -- Biological treatment Anaerobic bacteria Aerobic bacteria
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4011 , http://hdl.handle.net/10962/d1004071
- Description: 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.
- Full Text:
- Date Issued: 2004
- Authors: Ehlers, George A C
- Date: 2004
- Subjects: Hydrocarbons -- Biodegradation Sewage -- Purification -- Anaerobic treatment Water -- Purification -- Biological treatment Anaerobic bacteria Aerobic bacteria
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4011 , http://hdl.handle.net/10962/d1004071
- Description: 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.
- Full Text:
- Date Issued: 2004
The independent high rate algal pond as a unit operation in tertiary wastewater treatment
- Authors: Clark, Stewart James
- Date: 2002
- Subjects: Algae -- Biotechnology , Sewage -- Purification -- Biological treatment
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4092 , http://hdl.handle.net/10962/d1007805
- Description: The development of the High Rate Algal Pond (HRAP) as an independent tertiary treatment unit operation for phosphate and nitrate removal is reported. A novel Integrated Algal Ponding System (lAPS) design is proposed for nutrient removal from the effluents of both a conventional domestic sewage treatment plant and from an Advanced Integrated Wastewater Ponding System (AIWPS). The viability of an independently operated HRAP has been identified and termed the Independent High Rate Algal Pond (l-HRAP). A 500 m² pilot 1- HRAP was operated in such a way as to facilitate the precipitation of calcium phosphate, known to be controlled by pH (greater than 9.4) and resulting in final phosphate levels of less than 1 mg.L⁻¹ as P0₄-P. The incorporation of the I-HRAP into a denitrification process was also investigated. Continuously fed column reactors, utilising algal biomass as a carbon source, showed that the heterotrophic bacterial community dominant in the anaerobic algal sludge were denitrifying the nitrate in the feed. It was demonstrated that as the cultures were stressed (using increased nitrate concentrations, anaerobiosis and light starvation) total polysaccharide (TPS) concentrations increased, with a notable increase 111 the exopolysaccharide (EPS) fraction. These experiments corroborated the hypothesis that harvested microalgal biomass can be manipulated to produce, and release, exopolymeric substances under stress conditions, and which may serve as carbon source for denitrification. In both batch flask studies and in laboratory-scale reactor systems, harvested microalgal biomass from an HRAP was shown to produce exopolymeric substances under stress conditions. Initial high loading-rates of greater than 20 mg.L⁻¹ NO₃-N resulted in double the amount of exopolysaccharide production than in flasks with initial low loading-rates (less than 5 mg.L⁻¹ NO₃-N). Making use of an upflow anaerobic sludge blanket-type degrading-bed reactor, and an anaerobic, flooded trickle filter (ANTRIC) receiving HRAP effluent, the relationship between denitrification and the changes in polysaccharide content was investigated. This phenomenon has considerable beneficial implications in biological wastewater treatment systems where high nitrate concentration in the final effluent is a potential mitigating factor. Identification of the heterotrophic bacteria active in the denitrification process was attempted. This study presents a first report on the development and operation of the I-HRAP and has been followed by a technical-scale pilot plant evaluation of the process in the tertiary treatment of domestic wastewaters.
- Full Text:
- Date Issued: 2002
- Authors: Clark, Stewart James
- Date: 2002
- Subjects: Algae -- Biotechnology , Sewage -- Purification -- Biological treatment
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4092 , http://hdl.handle.net/10962/d1007805
- Description: The development of the High Rate Algal Pond (HRAP) as an independent tertiary treatment unit operation for phosphate and nitrate removal is reported. A novel Integrated Algal Ponding System (lAPS) design is proposed for nutrient removal from the effluents of both a conventional domestic sewage treatment plant and from an Advanced Integrated Wastewater Ponding System (AIWPS). The viability of an independently operated HRAP has been identified and termed the Independent High Rate Algal Pond (l-HRAP). A 500 m² pilot 1- HRAP was operated in such a way as to facilitate the precipitation of calcium phosphate, known to be controlled by pH (greater than 9.4) and resulting in final phosphate levels of less than 1 mg.L⁻¹ as P0₄-P. The incorporation of the I-HRAP into a denitrification process was also investigated. Continuously fed column reactors, utilising algal biomass as a carbon source, showed that the heterotrophic bacterial community dominant in the anaerobic algal sludge were denitrifying the nitrate in the feed. It was demonstrated that as the cultures were stressed (using increased nitrate concentrations, anaerobiosis and light starvation) total polysaccharide (TPS) concentrations increased, with a notable increase 111 the exopolysaccharide (EPS) fraction. These experiments corroborated the hypothesis that harvested microalgal biomass can be manipulated to produce, and release, exopolymeric substances under stress conditions, and which may serve as carbon source for denitrification. In both batch flask studies and in laboratory-scale reactor systems, harvested microalgal biomass from an HRAP was shown to produce exopolymeric substances under stress conditions. Initial high loading-rates of greater than 20 mg.L⁻¹ NO₃-N resulted in double the amount of exopolysaccharide production than in flasks with initial low loading-rates (less than 5 mg.L⁻¹ NO₃-N). Making use of an upflow anaerobic sludge blanket-type degrading-bed reactor, and an anaerobic, flooded trickle filter (ANTRIC) receiving HRAP effluent, the relationship between denitrification and the changes in polysaccharide content was investigated. This phenomenon has considerable beneficial implications in biological wastewater treatment systems where high nitrate concentration in the final effluent is a potential mitigating factor. Identification of the heterotrophic bacteria active in the denitrification process was attempted. This study presents a first report on the development and operation of the I-HRAP and has been followed by a technical-scale pilot plant evaluation of the process in the tertiary treatment of domestic wastewaters.
- Full Text:
- Date Issued: 2002
The molecular microbial ecology of sulfate reduction in the Rhodes BioSURE process
- Authors: Chauke, Chesa Gift
- Date: 2002
- Subjects: Water -- Purification -- Biological treatment , Acid mine drainage , Water -- Microbiology
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4082 , http://hdl.handle.net/10962/d1007475 , Water -- Purification -- Biological treatment , Acid mine drainage , Water -- Microbiology
- Description: 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.
- Full Text:
- Date Issued: 2002
- Authors: Chauke, Chesa Gift
- Date: 2002
- Subjects: Water -- Purification -- Biological treatment , Acid mine drainage , Water -- Microbiology
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4082 , http://hdl.handle.net/10962/d1007475 , Water -- Purification -- Biological treatment , Acid mine drainage , Water -- Microbiology
- Description: 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.
- Full Text:
- Date Issued: 2002
The Rhodes BioSure process in the treatment of acid mine drainage wastewaters
- Authors: Corbett, Christopher John
- Date: 2001 , 2013-05-03
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4077 , http://hdl.handle.net/10962/d1007405
- Description: While sulphate-enriched wastewaters are generated in a number of industrial processes, such as tanning, paper manufacture and metals processing, the principal contributors to large-scale pollution from this source in South Africa are the gold and coal mining industries. Both biological and physico-chemical processes, set in train by mining operations, give rise to the oxidation of sulphur species, and the resultant generation of AMD. The Vaal River system is most affected and receives large tonnages of mining related salinity as both direct discharges, and in diffuse runoff flows. The long-term burden of this problem, and sustaining ongoing treatment over the time-frames involved will almost certainly resort to the community inhabiting the area, notwithstanding progressive mine closure legislation and comprehensive regulation governing the polluterpays principle. The volume and time-frame of the AMD problem, and the need for a long-term and sustainable response has focused interest in biological treatment approaches. These have concentrated on active and passive treatment systems, both of which rely on microbial activity related to the biological sulphur cycle. Notwithstanding the reactor type, and the particular treatment approach used, widespread application of active AMD treatment has not yet been seen on any large scale. Singular factors constraining process development are bioreactor design, cost of bioreactor construction, and the cost of the carbon source and electron donor for the biological sulphate reduction process. The SRB are able to utilise only a limited range of small organic molecules. The studies reported here were motivated by the need to evaluate low-cost options and the treatment of high volume AMD flows. This has focussed research activity on bioprocess developments using complex organic compounds derived from waste streams as electron donor sources, and the integration of AMD treatment with other waste treatment objectives. The co-disposal of organic wastes with AMD treatment would enable the development of an 'integrated resource management' approach to the problem, including sustainability of treatment operations over the long time-frames involved. Apart from the cost advantages accrued to waste treatment, the recovery of the treated water as a resource to the wider community provides a potentially important value-added function to the combined operation. , KMBT_363 , Adobe Acrobat 9.53 Paper Capture Plug-in
- Full Text:
- Date Issued: 2001
- Authors: Corbett, Christopher John
- Date: 2001 , 2013-05-03
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4077 , http://hdl.handle.net/10962/d1007405
- Description: While sulphate-enriched wastewaters are generated in a number of industrial processes, such as tanning, paper manufacture and metals processing, the principal contributors to large-scale pollution from this source in South Africa are the gold and coal mining industries. Both biological and physico-chemical processes, set in train by mining operations, give rise to the oxidation of sulphur species, and the resultant generation of AMD. The Vaal River system is most affected and receives large tonnages of mining related salinity as both direct discharges, and in diffuse runoff flows. The long-term burden of this problem, and sustaining ongoing treatment over the time-frames involved will almost certainly resort to the community inhabiting the area, notwithstanding progressive mine closure legislation and comprehensive regulation governing the polluterpays principle. The volume and time-frame of the AMD problem, and the need for a long-term and sustainable response has focused interest in biological treatment approaches. These have concentrated on active and passive treatment systems, both of which rely on microbial activity related to the biological sulphur cycle. Notwithstanding the reactor type, and the particular treatment approach used, widespread application of active AMD treatment has not yet been seen on any large scale. Singular factors constraining process development are bioreactor design, cost of bioreactor construction, and the cost of the carbon source and electron donor for the biological sulphate reduction process. The SRB are able to utilise only a limited range of small organic molecules. The studies reported here were motivated by the need to evaluate low-cost options and the treatment of high volume AMD flows. This has focussed research activity on bioprocess developments using complex organic compounds derived from waste streams as electron donor sources, and the integration of AMD treatment with other waste treatment objectives. The co-disposal of organic wastes with AMD treatment would enable the development of an 'integrated resource management' approach to the problem, including sustainability of treatment operations over the long time-frames involved. Apart from the cost advantages accrued to waste treatment, the recovery of the treated water as a resource to the wider community provides a potentially important value-added function to the combined operation. , KMBT_363 , Adobe Acrobat 9.53 Paper Capture Plug-in
- Full Text:
- Date Issued: 2001
Development and characterisation of a membrane gradostat bioreactor for the bioremediation of aromatic pollutants using white rot fungi
- Authors: Leukes, Winston D
- Date: 1999
- Subjects: Aromatic compounds Pollutants Fungi Bioremediation Industrial microbiology Biotechnology
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4032 , http://hdl.handle.net/10962/d1004092
- Description: Bioremediation of aromatic pollutants using the ligninolytic enzymes of the white rot fungi has been thoroughly researched and has been shown to have considerable potential for industrial application. However, little success in scale-up and industrialisation of this technology has been attained due to problems associated with the continuous production of the pollutant-degrading enzymes using conventional bioreactor systems. The low productivities reported result from the incompatibility of conventional submerged culture reactor techniques with the physiological requirements of these fungi which have evolved on a solid-air interface, viz. wood. The enzymes are also produced only during the stationary phase of growth and can therefore be regarded as secondary metabolites. This study reports the conceptualisation, characterisation and evaluation of a novel bioreactor system as a solution to the continuous production of idiophasic pollutant degrading enzymes by the white rot fungus Phanerochaete chlysosporium. The reactor concept evolved from observation of these fungi in their native state, i. e. the metabolism of lignocellulosic material and involves the immobilisation of the organism onto a capillary ultrafiltration membrane. Nutrient gradients established across the biofilm, an inherent characteristic of fixed bed perfusion reactors, are exploited to provide both nutrient rich and nutrient poor zones across the biofilm. This allows growth or primary metabolism in the nutrient rich zone, pushing older biomass into the nutrient poor zone where secondary metabolism is induced by nutrient starvation. In effect, this represents a transformation of the events of a batch culture from a temporal to a spatial domain, allowing continuous production of secondary metabolites over time. Direct contact of the outer part of the biofilm with an air stream simulated the solid-air interface of the native state of the fungus. In order to facilitate the practical application of the membrane gradostat reactor (MGR) concept, conventional capillary membranes and membrane bioreactor modules were first evaluated. These were found to be unsuitable for application of the MGR concept. However, critical analysis of the shortcomings of the conventional systems resulted in the formulation of a set of design criteria for the development of a suitable membrane and module. These design criteria were satisfied by the development of a novel capillary membrane for membrane bioreactors, as well as a transverse flow membrane module, which is a novel approach in membrane bioreactor configuration. For the physiological characterisation of the MGR concept, a single fibre bioreactor unit was designed, which allowed destructive sampling of the biofilm for analysis. Using this system, it was shown that distinct morphological zones could be observed radially across the mature biofilm obtained through MGR operation. That these morphotypes do represent the temporal events of a typical batch culture in a spatial domain was confirmed by following the morphological changes occurring during batch culture of the immobilised fungus where the onset of primary and secondary metabolic conditions were manipulated through control of the nutrient supply. The different morphotypes were correlated to distinct growth phases by comparison of the morphology to the secretion of known enzymatic markers for secondary metabolism, viz. succinate dehydrogenase and cytochrome C oxidoreductase. Detailed structure-function analysis of the biofilm using transmission electron microscopy and adapted enzyme cytochemical staining techniques showed that the biofilm appeared to operate as a co-ordinated unit, with primary and secondary metabolism apparently linked in one thallus through nutrient translocation. This study provided new insights into the physiology of P. chrysosp,o rium and a detailed descriptive model was formulated which correlates well to existing models of wood degradation by the white rot fungi (WRF). Evaluation of the process on a laboratory scale using a novel transverse flow membrane bioreactor showed that a volumetric productivity of 1916 U.L.⁻¹day⁻¹ for manganese peroxidase, one of the pollutant degrading enzymes, could be attained, corresponding to a final concentration of 2 361 U.L.⁻¹ This may be compared to the best reported system (Moreira el at. 1997), where a volumetric productivity of 202 U.L.⁻¹day⁻¹was achieved with a final concentration of 250 U.L.⁻¹ However, MGR productivity is yet to be subjected to rigorous optimisation studies. The process could be operated continuously for 60 days. However, peak productivity could not be maintained for long periods. This was found to be due to physical phenomena relating to the fluid dynamics of the system which caused fluid flow maldistribution, which would have to be resolved through engineering analysis. In evaluation of the MGR concept for aromatic pollutant removal, in this case ρ- cresol, from growth medium, good performance was also achieved. The VmaxKm calculated by linear regression for the MGR was 0.8 (R² = 0.93), which compared favourably to that reported by Lewandowski et al. (1990), who obtained a Vmax/Km of 0.34 for a packed bed reactor treating chlorophenol. It was concluded that the MGR showed suitable potential to warrant further development, and that the descriptive characterisation of the biofilm physiology provided a sufficient basis for process analysis once engineering aspects ofthe system could be resolved.
- Full Text:
- Date Issued: 1999
- Authors: Leukes, Winston D
- Date: 1999
- Subjects: Aromatic compounds Pollutants Fungi Bioremediation Industrial microbiology Biotechnology
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4032 , http://hdl.handle.net/10962/d1004092
- Description: Bioremediation of aromatic pollutants using the ligninolytic enzymes of the white rot fungi has been thoroughly researched and has been shown to have considerable potential for industrial application. However, little success in scale-up and industrialisation of this technology has been attained due to problems associated with the continuous production of the pollutant-degrading enzymes using conventional bioreactor systems. The low productivities reported result from the incompatibility of conventional submerged culture reactor techniques with the physiological requirements of these fungi which have evolved on a solid-air interface, viz. wood. The enzymes are also produced only during the stationary phase of growth and can therefore be regarded as secondary metabolites. This study reports the conceptualisation, characterisation and evaluation of a novel bioreactor system as a solution to the continuous production of idiophasic pollutant degrading enzymes by the white rot fungus Phanerochaete chlysosporium. The reactor concept evolved from observation of these fungi in their native state, i. e. the metabolism of lignocellulosic material and involves the immobilisation of the organism onto a capillary ultrafiltration membrane. Nutrient gradients established across the biofilm, an inherent characteristic of fixed bed perfusion reactors, are exploited to provide both nutrient rich and nutrient poor zones across the biofilm. This allows growth or primary metabolism in the nutrient rich zone, pushing older biomass into the nutrient poor zone where secondary metabolism is induced by nutrient starvation. In effect, this represents a transformation of the events of a batch culture from a temporal to a spatial domain, allowing continuous production of secondary metabolites over time. Direct contact of the outer part of the biofilm with an air stream simulated the solid-air interface of the native state of the fungus. In order to facilitate the practical application of the membrane gradostat reactor (MGR) concept, conventional capillary membranes and membrane bioreactor modules were first evaluated. These were found to be unsuitable for application of the MGR concept. However, critical analysis of the shortcomings of the conventional systems resulted in the formulation of a set of design criteria for the development of a suitable membrane and module. These design criteria were satisfied by the development of a novel capillary membrane for membrane bioreactors, as well as a transverse flow membrane module, which is a novel approach in membrane bioreactor configuration. For the physiological characterisation of the MGR concept, a single fibre bioreactor unit was designed, which allowed destructive sampling of the biofilm for analysis. Using this system, it was shown that distinct morphological zones could be observed radially across the mature biofilm obtained through MGR operation. That these morphotypes do represent the temporal events of a typical batch culture in a spatial domain was confirmed by following the morphological changes occurring during batch culture of the immobilised fungus where the onset of primary and secondary metabolic conditions were manipulated through control of the nutrient supply. The different morphotypes were correlated to distinct growth phases by comparison of the morphology to the secretion of known enzymatic markers for secondary metabolism, viz. succinate dehydrogenase and cytochrome C oxidoreductase. Detailed structure-function analysis of the biofilm using transmission electron microscopy and adapted enzyme cytochemical staining techniques showed that the biofilm appeared to operate as a co-ordinated unit, with primary and secondary metabolism apparently linked in one thallus through nutrient translocation. This study provided new insights into the physiology of P. chrysosp,o rium and a detailed descriptive model was formulated which correlates well to existing models of wood degradation by the white rot fungi (WRF). Evaluation of the process on a laboratory scale using a novel transverse flow membrane bioreactor showed that a volumetric productivity of 1916 U.L.⁻¹day⁻¹ for manganese peroxidase, one of the pollutant degrading enzymes, could be attained, corresponding to a final concentration of 2 361 U.L.⁻¹ This may be compared to the best reported system (Moreira el at. 1997), where a volumetric productivity of 202 U.L.⁻¹day⁻¹was achieved with a final concentration of 250 U.L.⁻¹ However, MGR productivity is yet to be subjected to rigorous optimisation studies. The process could be operated continuously for 60 days. However, peak productivity could not be maintained for long periods. This was found to be due to physical phenomena relating to the fluid dynamics of the system which caused fluid flow maldistribution, which would have to be resolved through engineering analysis. In evaluation of the MGR concept for aromatic pollutant removal, in this case ρ- cresol, from growth medium, good performance was also achieved. The VmaxKm calculated by linear regression for the MGR was 0.8 (R² = 0.93), which compared favourably to that reported by Lewandowski et al. (1990), who obtained a Vmax/Km of 0.34 for a packed bed reactor treating chlorophenol. It was concluded that the MGR showed suitable potential to warrant further development, and that the descriptive characterisation of the biofilm physiology provided a sufficient basis for process analysis once engineering aspects ofthe system could be resolved.
- Full Text:
- Date Issued: 1999
The biotechnology of high rate algal ponding systems in the treatment of saline tannery wastewaters
- Authors: Dunn, Kevin Matthew
- Date: 1998
- Subjects: Sewage lagoons Tanneries -- Waste disposal Saline waters
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4006 , http://hdl.handle.net/10962/d1004066
- Description: Salinisation has been identified as a major cause of the progressive deterioration in the public water system in South Africa. To deal with this problem Waste Stabilisation Ponding systems have been used by the Leather Processing Industry as zero-dischaJ;ge wastewater evaporation disposal processes in water-limited inland regions of the country. While effective in the evaporation disposal function these systems are plagued by the generation of serious odour nuisance creating intractable environmental problems relating to adjacent residential communities. High loading to ponds of organic compounds, sulphides and ammonia results in strongly reducing anaerobic conditions prevailing in early parts of pond cascades. These are characterised by bright red colours due to the predominance of purple photosynthetic bacteria. Sporadic micro algal blooms of Spirulina sp. and Dunaliella sp. had been previously noted to occur on the latter ponds in these cascades, and were associated with their conversion to facultative function, with aerobic surface layers, and a marked reduction in odour release. This research programme undertook an investigation of the microbial ecology of a tannery waste stabilisation ponding system to describe factors which give rise to these blooms, and to determine whether microalgal growth may be manipulated to achieve a reliable oxygengenerating capping of the anaerobic ponds. The predominance of near pure cultures of Spin/lina platensis was demonstrated for the blooms and factors restricting its growth in the system were described. These include the interaction of ammonia and sulphide toxic effects and laboratory studies were undertaken to show how effluent loading may be regulated to enable effective growth of the cyanobacterium. At appropriate dilutions of tannery effluent an enhancement of growth was noted, compared to growth in defined mineral medium. An investigation of this phenomenon provided preliminary evidence for organic uptake by the pond micro algae and a possible contribution to heterotrophic nutrition. The manipulation of Spirulina sp. growth in a High Rate Algal Pond raceway was undertaken in outdoor pilot plant studies and the effect of microalgal capping of the anaerobic ponds in the cascade was demonstrated by activating a recycle loop from a blooming facultative pond. Heavy metal contaminants were effectively eliminated by an optimisation of the primary anaerobic pond function and precipitation as metal sulphides. Biomass was harvested and dried, during which a range of methods were evaluated. Toxicological studies were undertaken on the dried biomass using Artemia and chick assays, and feed studies showed its useful application in rations for the abalone Haliotlls midae and rainbow trout Onchorhynchlls mykiss. Based on positive independent assessment of research outcomes, a decision was made by the tanning company operating the Waste Stabilisation Ponding system, to proceed to the construction of a full-scale 2 500 m2 High Rate Algal Pond raceway. This would be used for controlled Spirlilina biomass production to effect a practical capping of the anaerobic ponds in the system, and to evaluate its commercial potential in the feed market. The Advanced Integrated Wastewater Ponding System described by Oswald (1991) provided the conceptual basis for the Algal Biotechnology process development undertaken. The studies of the microbial ecology and the biotechnological potential of this system have shown that a Spirulina-based High Rate Algal Ponding process can be engineered in such a way that saline tannery effluents may be treated to effect a significant reduction in overall pollution load, that biomass may be recovered as a value added product of the treatment process and that the operational performance of Waste Stabilisation Ponding systems, and hence their immediate environment, may be improved by the use of the High Rate Algal Pond as a retrofitted upgrading unit operation.
- Full Text:
- Date Issued: 1998
- Authors: Dunn, Kevin Matthew
- Date: 1998
- Subjects: Sewage lagoons Tanneries -- Waste disposal Saline waters
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4006 , http://hdl.handle.net/10962/d1004066
- Description: Salinisation has been identified as a major cause of the progressive deterioration in the public water system in South Africa. To deal with this problem Waste Stabilisation Ponding systems have been used by the Leather Processing Industry as zero-dischaJ;ge wastewater evaporation disposal processes in water-limited inland regions of the country. While effective in the evaporation disposal function these systems are plagued by the generation of serious odour nuisance creating intractable environmental problems relating to adjacent residential communities. High loading to ponds of organic compounds, sulphides and ammonia results in strongly reducing anaerobic conditions prevailing in early parts of pond cascades. These are characterised by bright red colours due to the predominance of purple photosynthetic bacteria. Sporadic micro algal blooms of Spirulina sp. and Dunaliella sp. had been previously noted to occur on the latter ponds in these cascades, and were associated with their conversion to facultative function, with aerobic surface layers, and a marked reduction in odour release. This research programme undertook an investigation of the microbial ecology of a tannery waste stabilisation ponding system to describe factors which give rise to these blooms, and to determine whether microalgal growth may be manipulated to achieve a reliable oxygengenerating capping of the anaerobic ponds. The predominance of near pure cultures of Spin/lina platensis was demonstrated for the blooms and factors restricting its growth in the system were described. These include the interaction of ammonia and sulphide toxic effects and laboratory studies were undertaken to show how effluent loading may be regulated to enable effective growth of the cyanobacterium. At appropriate dilutions of tannery effluent an enhancement of growth was noted, compared to growth in defined mineral medium. An investigation of this phenomenon provided preliminary evidence for organic uptake by the pond micro algae and a possible contribution to heterotrophic nutrition. The manipulation of Spirulina sp. growth in a High Rate Algal Pond raceway was undertaken in outdoor pilot plant studies and the effect of microalgal capping of the anaerobic ponds in the cascade was demonstrated by activating a recycle loop from a blooming facultative pond. Heavy metal contaminants were effectively eliminated by an optimisation of the primary anaerobic pond function and precipitation as metal sulphides. Biomass was harvested and dried, during which a range of methods were evaluated. Toxicological studies were undertaken on the dried biomass using Artemia and chick assays, and feed studies showed its useful application in rations for the abalone Haliotlls midae and rainbow trout Onchorhynchlls mykiss. Based on positive independent assessment of research outcomes, a decision was made by the tanning company operating the Waste Stabilisation Ponding system, to proceed to the construction of a full-scale 2 500 m2 High Rate Algal Pond raceway. This would be used for controlled Spirlilina biomass production to effect a practical capping of the anaerobic ponds in the system, and to evaluate its commercial potential in the feed market. The Advanced Integrated Wastewater Ponding System described by Oswald (1991) provided the conceptual basis for the Algal Biotechnology process development undertaken. The studies of the microbial ecology and the biotechnological potential of this system have shown that a Spirulina-based High Rate Algal Ponding process can be engineered in such a way that saline tannery effluents may be treated to effect a significant reduction in overall pollution load, that biomass may be recovered as a value added product of the treatment process and that the operational performance of Waste Stabilisation Ponding systems, and hence their immediate environment, may be improved by the use of the High Rate Algal Pond as a retrofitted upgrading unit operation.
- Full Text:
- Date Issued: 1998
Glycerol production by Dunaliella species in saline waste water treatment
- Authors: Emmett, Robyn Angela
- Date: 1996
- Subjects: Dunaliella -- Growth , Glycine
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4019 , http://hdl.handle.net/10962/d1004079 , Dunaliella -- Growth , Glycine
- Description: In recent years, much research has focused on Algal High Rate Oxidation Ponds as both an economic means for wastewater treatment and as a system for the mass production of algae. With the advent of these systems for the treatment of saline organic effluents, the extreme halophile, Dunaliella salina was considered. In this study, the growth and productivity of a number of Dunaliella species (and strains thereof) was evaluated in hide soak liquor tannery effluent. Hide soak liquor, diluted to 20% with water, proved to be highly suitable as a growth medium for the majority of the Dunaliella species under study and in some instances, resulted in enhanced growth rates and higher biomass yields compared to those obtained in defined inorganic medium. A few Dunaliella species failed to grow in this effluent. A correlation was observed between the lack of growth displayed by these species in this organio-rich medium and their failure to utilise organic compounds. Glycine, a major component of this effluent, possibly stimulates the growth of Dunaliella. Studies on the mechanism of growth stimulation by glycine revealed that an algal-bacterial relationship existed whereby the bacteria mineralised the amino acid, releasing ammonia which was then utilised by the alga. Results of this work revealed significant variations in the intracellular glycerol content amongst the Dunaliella species under study. Large differences were also observed between the glycerol contents of effluent-grown and control Dunaliella cells, where the effluent-grown cells were characterised by greatly reduced intracellular glycerol content. These reduced glycerol levels are assumed to have arisen from the glycine-induced stimulation of glycerol release which was observed in this study, where the high glycine content of the hide soak liquor is proposed to have induced glycerol release. This enhanced glycerol release in tatmery effluent could play a central role in the fimction of Dunaliella-based High Rate Oxidation Ponding systems, by stimulating bacterial activity. Observed glycerol productivities were therefore proposed to be a fimction of the type and concentration of the organic constituents of the medium. A similar medium-induced phenomenon was observed in the starch content of Dunaliella cells.
- Full Text:
- Date Issued: 1996
- Authors: Emmett, Robyn Angela
- Date: 1996
- Subjects: Dunaliella -- Growth , Glycine
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4019 , http://hdl.handle.net/10962/d1004079 , Dunaliella -- Growth , Glycine
- Description: In recent years, much research has focused on Algal High Rate Oxidation Ponds as both an economic means for wastewater treatment and as a system for the mass production of algae. With the advent of these systems for the treatment of saline organic effluents, the extreme halophile, Dunaliella salina was considered. In this study, the growth and productivity of a number of Dunaliella species (and strains thereof) was evaluated in hide soak liquor tannery effluent. Hide soak liquor, diluted to 20% with water, proved to be highly suitable as a growth medium for the majority of the Dunaliella species under study and in some instances, resulted in enhanced growth rates and higher biomass yields compared to those obtained in defined inorganic medium. A few Dunaliella species failed to grow in this effluent. A correlation was observed between the lack of growth displayed by these species in this organio-rich medium and their failure to utilise organic compounds. Glycine, a major component of this effluent, possibly stimulates the growth of Dunaliella. Studies on the mechanism of growth stimulation by glycine revealed that an algal-bacterial relationship existed whereby the bacteria mineralised the amino acid, releasing ammonia which was then utilised by the alga. Results of this work revealed significant variations in the intracellular glycerol content amongst the Dunaliella species under study. Large differences were also observed between the glycerol contents of effluent-grown and control Dunaliella cells, where the effluent-grown cells were characterised by greatly reduced intracellular glycerol content. These reduced glycerol levels are assumed to have arisen from the glycine-induced stimulation of glycerol release which was observed in this study, where the high glycine content of the hide soak liquor is proposed to have induced glycerol release. This enhanced glycerol release in tatmery effluent could play a central role in the fimction of Dunaliella-based High Rate Oxidation Ponding systems, by stimulating bacterial activity. Observed glycerol productivities were therefore proposed to be a fimction of the type and concentration of the organic constituents of the medium. A similar medium-induced phenomenon was observed in the starch content of Dunaliella cells.
- Full Text:
- Date Issued: 1996
Physiological signal transduction from the photosynthetic apparatus in the green alga Dunaliella salina
- Logie, Malcolme Ronald Ruxton
- Authors: Logie, Malcolme Ronald Ruxton
- Date: 1995
- Subjects: Cellular signal transduction Photosynthesis -- Research Green algae Dunaliella
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4035 , http://hdl.handle.net/10962/d1004095
- Description: The transduction of stress signals in plants is known to involve complex hysiological responses. In D. salina a range of stresses results in hyperaccumulation of ft-carotene and an understanding of stress responses in this organism has important biotechnological implications. In this thesis an attempt was made to elucidate the physiological components involved and establish a role for pH in response to high light stress. In order to achieve this the effect of high light stress on photosynthesis and cell productivity was measured. Results showed that photosynthetic carbon assimilation, oxygen evolution and cellular productivity was initially inhibited by exposure to high light intensities, but this inhibition was transient and was overcome by a rapid increase in all three parameters. The response of the carbon pool intermediates was also investigated. It was shown that on exposure to high light ft-carotene declined but then showed a rapid increase after about 4 hours of exposure. It was also demonstrated that the initial loss of ft-carotene was due to loss of this pigment from the photosynthetic pigment bed and that the hyper-accumulation of ft-carotene was due to accumulation of ft-carotene in lipoidal globules located in the chloroplast stroma. It was further demonstrated that there was mass movement of carbon in the xanthophyll cycle shortly after exposure to high light. This was characterized by the de-epoxidation of violaxanthin to antheraxanthin with a further de-epoxidation to zeaxanthin, thereby decreasing the epoxidation state of the cycle. Furthermore, it was shown that there was relocation of carbon from violaxanthin to the plant growth regulator abscisic acid. It was also shown for the first time in D. salina that the production of ft-carotene and operation of the epoxidation state of the xanthophyll cycle has a periodicity which is established after exposure to successive cycles of a light regime. Chlorophyll fluorescence was used together with well established ammonia stress responses to acquire a general overview of energy dissipation from the photosynthetic pigment bed. In conjunction with an understanding of xanthophyll cycle operation during exposure to high light stress it has been possible to establish a relationship between chlorophyll florescence, xanthophyll cycle operation and intracellular pH. It was also shown using chlorophyll fluorescence that after 4 hour exposure to high light a maximum fluorescence peak could no longer be induced indicating a transition at about this point from a state of reversibility to commitment of the full stress response. Nuclear magnetic resonance was used to follow intracellular pH fluxes during exposure to high light. A novel technique was developed for studying photosynthetically active organisms in the dark using nuclear magnetic resonance. These results showed that on exposure to high light stress there is rapid acidification of the chloroplast stroma and to a lesser degree of the acidic vacuole. The pH of these compartments is re-established after about 4 hours which is co-incident with the onset of fl-carotene hyper-accumulation and the loss of the induction of the chlorophyll fluorescence peak indicating an intimate relationship for fl-carotene, chlorophyll fluorescence, xanthophyll cycle operation and pH. The results from this study allow for the proposal of a general physiological stress transduction response mechanism for D. salina which is common for a range of different stresses and where intracellular pH plays a central role.
- Full Text:
- Date Issued: 1995
- Authors: Logie, Malcolme Ronald Ruxton
- Date: 1995
- Subjects: Cellular signal transduction Photosynthesis -- Research Green algae Dunaliella
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4035 , http://hdl.handle.net/10962/d1004095
- Description: The transduction of stress signals in plants is known to involve complex hysiological responses. In D. salina a range of stresses results in hyperaccumulation of ft-carotene and an understanding of stress responses in this organism has important biotechnological implications. In this thesis an attempt was made to elucidate the physiological components involved and establish a role for pH in response to high light stress. In order to achieve this the effect of high light stress on photosynthesis and cell productivity was measured. Results showed that photosynthetic carbon assimilation, oxygen evolution and cellular productivity was initially inhibited by exposure to high light intensities, but this inhibition was transient and was overcome by a rapid increase in all three parameters. The response of the carbon pool intermediates was also investigated. It was shown that on exposure to high light ft-carotene declined but then showed a rapid increase after about 4 hours of exposure. It was also demonstrated that the initial loss of ft-carotene was due to loss of this pigment from the photosynthetic pigment bed and that the hyper-accumulation of ft-carotene was due to accumulation of ft-carotene in lipoidal globules located in the chloroplast stroma. It was further demonstrated that there was mass movement of carbon in the xanthophyll cycle shortly after exposure to high light. This was characterized by the de-epoxidation of violaxanthin to antheraxanthin with a further de-epoxidation to zeaxanthin, thereby decreasing the epoxidation state of the cycle. Furthermore, it was shown that there was relocation of carbon from violaxanthin to the plant growth regulator abscisic acid. It was also shown for the first time in D. salina that the production of ft-carotene and operation of the epoxidation state of the xanthophyll cycle has a periodicity which is established after exposure to successive cycles of a light regime. Chlorophyll fluorescence was used together with well established ammonia stress responses to acquire a general overview of energy dissipation from the photosynthetic pigment bed. In conjunction with an understanding of xanthophyll cycle operation during exposure to high light stress it has been possible to establish a relationship between chlorophyll florescence, xanthophyll cycle operation and intracellular pH. It was also shown using chlorophyll fluorescence that after 4 hour exposure to high light a maximum fluorescence peak could no longer be induced indicating a transition at about this point from a state of reversibility to commitment of the full stress response. Nuclear magnetic resonance was used to follow intracellular pH fluxes during exposure to high light. A novel technique was developed for studying photosynthetically active organisms in the dark using nuclear magnetic resonance. These results showed that on exposure to high light stress there is rapid acidification of the chloroplast stroma and to a lesser degree of the acidic vacuole. The pH of these compartments is re-established after about 4 hours which is co-incident with the onset of fl-carotene hyper-accumulation and the loss of the induction of the chlorophyll fluorescence peak indicating an intimate relationship for fl-carotene, chlorophyll fluorescence, xanthophyll cycle operation and pH. The results from this study allow for the proposal of a general physiological stress transduction response mechanism for D. salina which is common for a range of different stresses and where intracellular pH plays a central role.
- Full Text:
- Date Issued: 1995
A process for the detanning of chrome leather wastes utilising tannery effluents
- Authors: Glaum, Deanne Melanie
- Date: 1994
- Subjects: Tanneries -- Waste disposal , Recycling (Waste, etc.) , Leather
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4029 , http://hdl.handle.net/10962/d1004089 , Tanneries -- Waste disposal , Recycling (Waste, etc.) , Leather
- Description: The considerable volume of chromium-bearing wastes generated during the process of leather tanning, exacerbated by the potential for trivalent chromium in the wastes to be oxidised to the toxic hexavalent state, has created a major waste disposal dilemma for the tanning industry. While methods are available for the safe and effective treatment of residual chrome-tanning liquors, little has been done to address the issue of the chrome-bearing solid wastes. Given the increasingly stringent environmental compliance standards facing tanneries, unless an appropriate treatment process is developed in the immediate future, the continued use of chromium as a tanning agent could be compromised. Recent investigations have demonstrated the potential of heated alkaline conditions for dechroming these solid wastes. This study expanded upon these considerations and examined the feasibility of utilising the highly alkaline tannery waste effluents as cost-effective, substitute alkaline media. The three effluents considered in this study, classed as lime sulphide liquors, were shown to be capable of dechroming wet blue shavings, with resultant separation of the solid wastes into a protein and a concentrated chromium product. The solubilised protein product contained low chromium concentrations which comply with legal discharge limits. The precipitated chromium product offers opportunity for reutilisation in the tannery. A novel industrial-scale treatment process, based on these investigations, indicated the process to be capable of treating the quantity of shavings produced on a daily basis by a medium to large scale tannery. Application of this method for the dechroming of other chrome-tanned solid wastes was also shown to be feasible.
- Full Text:
- Date Issued: 1994
- Authors: Glaum, Deanne Melanie
- Date: 1994
- Subjects: Tanneries -- Waste disposal , Recycling (Waste, etc.) , Leather
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4029 , http://hdl.handle.net/10962/d1004089 , Tanneries -- Waste disposal , Recycling (Waste, etc.) , Leather
- Description: The considerable volume of chromium-bearing wastes generated during the process of leather tanning, exacerbated by the potential for trivalent chromium in the wastes to be oxidised to the toxic hexavalent state, has created a major waste disposal dilemma for the tanning industry. While methods are available for the safe and effective treatment of residual chrome-tanning liquors, little has been done to address the issue of the chrome-bearing solid wastes. Given the increasingly stringent environmental compliance standards facing tanneries, unless an appropriate treatment process is developed in the immediate future, the continued use of chromium as a tanning agent could be compromised. Recent investigations have demonstrated the potential of heated alkaline conditions for dechroming these solid wastes. This study expanded upon these considerations and examined the feasibility of utilising the highly alkaline tannery waste effluents as cost-effective, substitute alkaline media. The three effluents considered in this study, classed as lime sulphide liquors, were shown to be capable of dechroming wet blue shavings, with resultant separation of the solid wastes into a protein and a concentrated chromium product. The solubilised protein product contained low chromium concentrations which comply with legal discharge limits. The precipitated chromium product offers opportunity for reutilisation in the tannery. A novel industrial-scale treatment process, based on these investigations, indicated the process to be capable of treating the quantity of shavings produced on a daily basis by a medium to large scale tannery. Application of this method for the dechroming of other chrome-tanned solid wastes was also shown to be feasible.
- Full Text:
- Date Issued: 1994
Biocatalytic and biomimetic studies of polyphenol oxidase
- Authors: Burton, Stephanie Gail
- Date: 1994
- Subjects: Phenol oxidase Polyphenols Oxidases
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4028 , http://hdl.handle.net/10962/d1004088
- Description: Mushroom polyphenol oxidase (EC 1.14.18.1) was investigated to determine its potential for application as a biocatalyst in the synthesis of o-quinones, in organic medium. In order to determine the kinetic properties of the biocatalyst, a system was devised which comprised an immobilised polyphenol oxidase extract, functioning in chloroform. The system was hydrated by the addition of buffer. A simple method for the consistent measurement of reaction rates in this heterogenous system was designed and used to obtain detailed enzyme kinetic data relating to optimisation of reaction conditions and substrate specificity. The aqueous content of the system was optimised using p-cresol as a substrate. A crude, immobilised extract of Agaricus bisporus was used to hydroxylate and oxidise a range of selected p-substituted phenolic substrates, yielding, as the sale products, o-quinones. These products were efficiently reduced to catechols by extracting the reaction mixtures with aqueous ascorbic acid solution. The biocatalytic system was also successfully utilised to produce L-DOPA, the drug used to treat Parkinson's disease, from L-acetyl tyrosine ethyl ester (ATEE). Michaelis-Menten kinetics were used to obtain apparent Km and V values with respect to the selected phenolic substrates, and the kinetic parameters obtained were found to correlate well with the steric requirements of the substrates and with their hydrophobicity. In the course of the investigation, a novel ¹H NMR method was used to facilitate measurement of the UV molar absorption coefficients of the o-quinones in reaction mixtures, thus avoiding the necessity to isolate these unstable, water-sensitive products. The biocatalytic system was extended to a continuous process, in which the immobilised enzyme was shown to function successfully in the chloroform medium for several hours, with high conversion rates. Modifications, involving partial purification and the addition of a surfactant, were investigated to determine their effect on the kinetic parameters. The results obtained using partially purified enzyme indicated that the removal of extraneous protein and/or melanoid material lead to a reduced capacity for conversion of sterically demanding substrates. The addition of the anionic detergent, sodium dodecyl sulphate (SOS), enhanced the ability of the biocatalyst to bind and oxidise sterically demanding substrates. These effects are attributed to changes in the polar state of groups within the protein binding pocket, which result in altered flexibility and hydrophobicity. Computer modelling of several biomimetic dinuclear copper complexes also indicated the importance of flexibility for effective biocatalysis. Novel binuclear copper (II complexes, containing a flexible biphenyl spacer and imidazole or benzimidazole donors, were prepared and analysed using NMR, UV, AA and cyclic voltammetric techniques. The complexes were also shown, in a detailed kinetic study, to mimic the catecholase activity of polyphenol oxidase by oxidising 3,5-di-tertbutylcatechol, and to catalyse the coupling of the phenolic substrate 2,4-di-tert-butylphenol. However, the complexes were apparently too flexible to react with smaller substrates. These biomimetic complexes provided valuable insights into the nature of the dinuclear copper binding site.
- Full Text:
- Date Issued: 1994
- Authors: Burton, Stephanie Gail
- Date: 1994
- Subjects: Phenol oxidase Polyphenols Oxidases
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4028 , http://hdl.handle.net/10962/d1004088
- Description: Mushroom polyphenol oxidase (EC 1.14.18.1) was investigated to determine its potential for application as a biocatalyst in the synthesis of o-quinones, in organic medium. In order to determine the kinetic properties of the biocatalyst, a system was devised which comprised an immobilised polyphenol oxidase extract, functioning in chloroform. The system was hydrated by the addition of buffer. A simple method for the consistent measurement of reaction rates in this heterogenous system was designed and used to obtain detailed enzyme kinetic data relating to optimisation of reaction conditions and substrate specificity. The aqueous content of the system was optimised using p-cresol as a substrate. A crude, immobilised extract of Agaricus bisporus was used to hydroxylate and oxidise a range of selected p-substituted phenolic substrates, yielding, as the sale products, o-quinones. These products were efficiently reduced to catechols by extracting the reaction mixtures with aqueous ascorbic acid solution. The biocatalytic system was also successfully utilised to produce L-DOPA, the drug used to treat Parkinson's disease, from L-acetyl tyrosine ethyl ester (ATEE). Michaelis-Menten kinetics were used to obtain apparent Km and V values with respect to the selected phenolic substrates, and the kinetic parameters obtained were found to correlate well with the steric requirements of the substrates and with their hydrophobicity. In the course of the investigation, a novel ¹H NMR method was used to facilitate measurement of the UV molar absorption coefficients of the o-quinones in reaction mixtures, thus avoiding the necessity to isolate these unstable, water-sensitive products. The biocatalytic system was extended to a continuous process, in which the immobilised enzyme was shown to function successfully in the chloroform medium for several hours, with high conversion rates. Modifications, involving partial purification and the addition of a surfactant, were investigated to determine their effect on the kinetic parameters. The results obtained using partially purified enzyme indicated that the removal of extraneous protein and/or melanoid material lead to a reduced capacity for conversion of sterically demanding substrates. The addition of the anionic detergent, sodium dodecyl sulphate (SOS), enhanced the ability of the biocatalyst to bind and oxidise sterically demanding substrates. These effects are attributed to changes in the polar state of groups within the protein binding pocket, which result in altered flexibility and hydrophobicity. Computer modelling of several biomimetic dinuclear copper complexes also indicated the importance of flexibility for effective biocatalysis. Novel binuclear copper (II complexes, containing a flexible biphenyl spacer and imidazole or benzimidazole donors, were prepared and analysed using NMR, UV, AA and cyclic voltammetric techniques. The complexes were also shown, in a detailed kinetic study, to mimic the catecholase activity of polyphenol oxidase by oxidising 3,5-di-tertbutylcatechol, and to catalyse the coupling of the phenolic substrate 2,4-di-tert-butylphenol. However, the complexes were apparently too flexible to react with smaller substrates. These biomimetic complexes provided valuable insights into the nature of the dinuclear copper binding site.
- Full Text:
- Date Issued: 1994
Stress manipulation in Dunaliella salina and dual-stage [beta]-carotene production
- Authors: Phillips, Trevor David
- Date: 1994
- Subjects: Dunaliella Carotenes Plants -- Effect of stress on
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4037 , http://hdl.handle.net/10962/d1004097
- Description: The alga Dunaliella salina accumulates large quantities of β-carotene in response to certain environmental and physiological stresses. This hyper-accumulation process has been commercially exploited. However, the currently employed averaging or single-stage process produces β-carotene yields well below the genetic potential of the organism due to the inverse relationship between growth and secondary metabolite production. A dual-stage process, which separates the distinctive growth and secondary metabolite production stages of the alga, has been proposed. The broad aim of the research programme was to evaluate the practicality, scale-up and economic viability of a dual-stage β-carotene production process from D. salina. Preliminary laboratory studies showed that although stress factors such as high salinity and a range of nutrient limitations enhance β-carotene accumulation in D. salina, high light intensity is the single most important factor inducing β-carotene hyper-accumulation in the alga. Furthermore, the preliminary studies indicated that 6-carotene production could be successfully manipulated by the imposition of stress. The stress response of D. salina to high light stress was examined at a fundamental level. The relative partitioning of β-carotene between thylakoid membrane and interthylakoid globular β-carotene has revealed two responses to high light stress. The first is a response in which the alga adapts to the photoinhibitory effects of high light stress by the rapid accumulation and the peripheral localisation of Jl-carotene to the outer extremities of the chloroplast. This is followed by a maintenance response which is characterised by the recovery of the photosynthetic rate and cell growth. A possible interrelationship between the extent of the photo inhibitory response and the amount of β-carotene hyper-accumulation has been noted. An outdoor evaluation of the growth stage of the dual-stage system has demonstrated that D. salina can be grown in a relatively low salinity, nutrient sufficient medium for extended periods without overgrowth by small non-carotenogenic Dunaliella species. In addition, biomass productivities of three times greater than those obtained in the currently employed averaging system were achieved. The role of high light intensity in β-carotene hyper-accumulation was confirmed in outdoor scale-up stress pond studies. The studies demonstrated the feasibility of stress induced ll-carotene production in outdoor cultures of D. salina and β-carotene yields three times greater than those obtained in the currently employed averaging process were achieved. The dual-stage process imposes the specific requirement of viable cell separation on the harvesting system employed. A flocculation-flotation process and an air-displacement crossflow ultrafiltration system were developed and successfully evaluated for the separation of D. salina from the brine solution in a viable form. The extraction of β-carotene from D. salina was evaluated. Supercritical fluid extraction studies showed that the use of a co-solvent mixture of carbon dioxide and propane could effectively reduce the high extraction pressures associated with supercritical carbon dioxide extraction. In addition, a novel hydrophobic membrane assisted hot oil extraction process was developed which separates the complex oil-water emulsions produced during hot oil extraction of 6-carotene from wet D. salina biomass. Process design and economic evaluation studies were undertaken and showed that the economics of the dual-stage process offer significant advantages over the currently employed averaging process.
- Full Text:
- Date Issued: 1994
- Authors: Phillips, Trevor David
- Date: 1994
- Subjects: Dunaliella Carotenes Plants -- Effect of stress on
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4037 , http://hdl.handle.net/10962/d1004097
- Description: The alga Dunaliella salina accumulates large quantities of β-carotene in response to certain environmental and physiological stresses. This hyper-accumulation process has been commercially exploited. However, the currently employed averaging or single-stage process produces β-carotene yields well below the genetic potential of the organism due to the inverse relationship between growth and secondary metabolite production. A dual-stage process, which separates the distinctive growth and secondary metabolite production stages of the alga, has been proposed. The broad aim of the research programme was to evaluate the practicality, scale-up and economic viability of a dual-stage β-carotene production process from D. salina. Preliminary laboratory studies showed that although stress factors such as high salinity and a range of nutrient limitations enhance β-carotene accumulation in D. salina, high light intensity is the single most important factor inducing β-carotene hyper-accumulation in the alga. Furthermore, the preliminary studies indicated that 6-carotene production could be successfully manipulated by the imposition of stress. The stress response of D. salina to high light stress was examined at a fundamental level. The relative partitioning of β-carotene between thylakoid membrane and interthylakoid globular β-carotene has revealed two responses to high light stress. The first is a response in which the alga adapts to the photoinhibitory effects of high light stress by the rapid accumulation and the peripheral localisation of Jl-carotene to the outer extremities of the chloroplast. This is followed by a maintenance response which is characterised by the recovery of the photosynthetic rate and cell growth. A possible interrelationship between the extent of the photo inhibitory response and the amount of β-carotene hyper-accumulation has been noted. An outdoor evaluation of the growth stage of the dual-stage system has demonstrated that D. salina can be grown in a relatively low salinity, nutrient sufficient medium for extended periods without overgrowth by small non-carotenogenic Dunaliella species. In addition, biomass productivities of three times greater than those obtained in the currently employed averaging system were achieved. The role of high light intensity in β-carotene hyper-accumulation was confirmed in outdoor scale-up stress pond studies. The studies demonstrated the feasibility of stress induced ll-carotene production in outdoor cultures of D. salina and β-carotene yields three times greater than those obtained in the currently employed averaging process were achieved. The dual-stage process imposes the specific requirement of viable cell separation on the harvesting system employed. A flocculation-flotation process and an air-displacement crossflow ultrafiltration system were developed and successfully evaluated for the separation of D. salina from the brine solution in a viable form. The extraction of β-carotene from D. salina was evaluated. Supercritical fluid extraction studies showed that the use of a co-solvent mixture of carbon dioxide and propane could effectively reduce the high extraction pressures associated with supercritical carbon dioxide extraction. In addition, a novel hydrophobic membrane assisted hot oil extraction process was developed which separates the complex oil-water emulsions produced during hot oil extraction of 6-carotene from wet D. salina biomass. Process design and economic evaluation studies were undertaken and showed that the economics of the dual-stage process offer significant advantages over the currently employed averaging process.
- Full Text:
- Date Issued: 1994
The biotechnology of effluent-grown Spirulina, and application in aquaculture nutrition
- Authors: Maart, Brenton Ashley
- Date: 1993
- Subjects: Aquaculture , Spirulina , Algae -- Biotechnology , Fishes -- Feeding and feeds
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4050 , http://hdl.handle.net/10962/d1004111 , Aquaculture , Spirulina , Algae -- Biotechnology , Fishes -- Feeding and feeds
- Description: The biotechnology of production and utilisation of the cyanobacterium Spirulina has been well documented. Research has centred mainly on application in human and animal nutrition, and has been motivated by the high protein, vitamin, fatty acid and growth factor contents. The main obstacle in realising the full potential of this feed source has been the high production costs associated with its mass culture in defined media. The observation of blooms of Spirulina in tannery effluent evaporation ponds in Wellington, South Africa, prompted this investigation into the harvesting, and nutritional and toxicological evaluation of this potentially low-cost production system, with the ultimate aim of using the product in aquaculture rations. An investigation of the chemical gradient along the evaporation cascade showed a positive correlation between the prevailing chemical conditions and the dominant species populations. A standing crop of 9.5 tonnes/ha of Spirulina was found to be present in the latter alkaline ponds, characterised by relatively lower organic and sulphur contents. Initial harvesting of the biomass was achieved by the design, construction and implementation of a small-scale screen harvest, which yielded a 25 kg (dry weight) crop. A scale-up model was then designed, and implemented in a technical scale harvest, yielding a crop of 250 kg (dry weight). Both these harvests utilised the bloom of surface-autoflocculated biomass. Concentrated cell slurries were sun-dried on muslin beds, and milled to a coarse powder. An evaluation of the harvest revealed a chemical content similar to other published reports of defined media cultures, with the exception of the protein and amino acid contents. The observed lower levels of the latter two are almost certainly due to the sun-drying method employed, known to reduce the protein content due to thermal denaturation. Legislation demands the strict toxicological evaluation of new protein sources, and because of the effluent-nature of the growth medium of this source of Spirulina, its viability lies only in the application as an animal feed or supplement. A range of toxicological tests were chosen that were targeted to elucidate the possible toxicological constraints of this effluentgrown source of protein in animal nutrition. The nucleic acid and pesticide contents of the harvested biomass were within the prescribed safety ranges. Atomic absorption showed minimal accumulation of minerals and heavy metals from the effluent. A bioassay with the brine shrimp Anemia salina showed that the biomass contained no toxicologically active water-soluble components. A short term feeding trial with new-born chicks showed that supplementation with Spirulina had no effect on the growth rates and feed conversion ratios of the different feeding groups. Pathological analyses showed that the liver was the only target organ to elicit a change in response to supplementation of the diets with Spirulina. A general decrease in liver weight was noted, with Cu, Ca, Fe and Zn being significantly accumulated. A histopathological examination however, showed no cellular and functional aberration from the control animals. The toxicological analyses gave the preliminary safe go-ahead for the evaluation of effluent-grown Spirulina in aquaculture nutrition. The South African abalone Haliotis midae, and the rainbow trout Oncorhynchus mykiss were chosen as representative species of edible cultured organisms. The technology for the culture of the perlemoen abalone is being established in South Africa, with the main area of research being the development of an artificial diet for high density culture. A 40 day growth trial demonstrated that lower concentrations of Spirulina supplemented to an agar-based fishmeal diet resulted in growth rates and feed conversion ratios similar to the control fishmeal and purified-casein diets, and thus has application potential in the nutrition of this high-cost marine delicacy. The aquaculture technology of freshwater rainbow trout is already well established. An eight week feeding trial with various concentrations of Spirulina showed that this effluent-grown protein source can partially replace fishmeal in semi-purified diets. Fish fed Spirulina did not exhibit decisive manifestations of toxicity, as determined in a histopathological study. In addition, Spirulina supplementation resulted in enhanced colouration of the skin and flesh, which may have implications in the aesthetic marketing of this sought-after table fish. The primary aim of this preliminary investigation thus concerned the determination of the biotechnological potential of this effluent-source of Spirulina. A technology transfer from the economically unfeasible defined-media culture was implemented. This project is ultimately aimed as a contribution towards the treatment of tannery wastewater, by the removal of contaminants from the effluent in the form of organic biomass.
- Full Text:
- Date Issued: 1993
- Authors: Maart, Brenton Ashley
- Date: 1993
- Subjects: Aquaculture , Spirulina , Algae -- Biotechnology , Fishes -- Feeding and feeds
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4050 , http://hdl.handle.net/10962/d1004111 , Aquaculture , Spirulina , Algae -- Biotechnology , Fishes -- Feeding and feeds
- Description: The biotechnology of production and utilisation of the cyanobacterium Spirulina has been well documented. Research has centred mainly on application in human and animal nutrition, and has been motivated by the high protein, vitamin, fatty acid and growth factor contents. The main obstacle in realising the full potential of this feed source has been the high production costs associated with its mass culture in defined media. The observation of blooms of Spirulina in tannery effluent evaporation ponds in Wellington, South Africa, prompted this investigation into the harvesting, and nutritional and toxicological evaluation of this potentially low-cost production system, with the ultimate aim of using the product in aquaculture rations. An investigation of the chemical gradient along the evaporation cascade showed a positive correlation between the prevailing chemical conditions and the dominant species populations. A standing crop of 9.5 tonnes/ha of Spirulina was found to be present in the latter alkaline ponds, characterised by relatively lower organic and sulphur contents. Initial harvesting of the biomass was achieved by the design, construction and implementation of a small-scale screen harvest, which yielded a 25 kg (dry weight) crop. A scale-up model was then designed, and implemented in a technical scale harvest, yielding a crop of 250 kg (dry weight). Both these harvests utilised the bloom of surface-autoflocculated biomass. Concentrated cell slurries were sun-dried on muslin beds, and milled to a coarse powder. An evaluation of the harvest revealed a chemical content similar to other published reports of defined media cultures, with the exception of the protein and amino acid contents. The observed lower levels of the latter two are almost certainly due to the sun-drying method employed, known to reduce the protein content due to thermal denaturation. Legislation demands the strict toxicological evaluation of new protein sources, and because of the effluent-nature of the growth medium of this source of Spirulina, its viability lies only in the application as an animal feed or supplement. A range of toxicological tests were chosen that were targeted to elucidate the possible toxicological constraints of this effluentgrown source of protein in animal nutrition. The nucleic acid and pesticide contents of the harvested biomass were within the prescribed safety ranges. Atomic absorption showed minimal accumulation of minerals and heavy metals from the effluent. A bioassay with the brine shrimp Anemia salina showed that the biomass contained no toxicologically active water-soluble components. A short term feeding trial with new-born chicks showed that supplementation with Spirulina had no effect on the growth rates and feed conversion ratios of the different feeding groups. Pathological analyses showed that the liver was the only target organ to elicit a change in response to supplementation of the diets with Spirulina. A general decrease in liver weight was noted, with Cu, Ca, Fe and Zn being significantly accumulated. A histopathological examination however, showed no cellular and functional aberration from the control animals. The toxicological analyses gave the preliminary safe go-ahead for the evaluation of effluent-grown Spirulina in aquaculture nutrition. The South African abalone Haliotis midae, and the rainbow trout Oncorhynchus mykiss were chosen as representative species of edible cultured organisms. The technology for the culture of the perlemoen abalone is being established in South Africa, with the main area of research being the development of an artificial diet for high density culture. A 40 day growth trial demonstrated that lower concentrations of Spirulina supplemented to an agar-based fishmeal diet resulted in growth rates and feed conversion ratios similar to the control fishmeal and purified-casein diets, and thus has application potential in the nutrition of this high-cost marine delicacy. The aquaculture technology of freshwater rainbow trout is already well established. An eight week feeding trial with various concentrations of Spirulina showed that this effluent-grown protein source can partially replace fishmeal in semi-purified diets. Fish fed Spirulina did not exhibit decisive manifestations of toxicity, as determined in a histopathological study. In addition, Spirulina supplementation resulted in enhanced colouration of the skin and flesh, which may have implications in the aesthetic marketing of this sought-after table fish. The primary aim of this preliminary investigation thus concerned the determination of the biotechnological potential of this effluent-source of Spirulina. A technology transfer from the economically unfeasible defined-media culture was implemented. This project is ultimately aimed as a contribution towards the treatment of tannery wastewater, by the removal of contaminants from the effluent in the form of organic biomass.
- Full Text:
- Date Issued: 1993
The microbial production of polyphenol oxidase enzyme systems and their application in the treatment of phenolic wastewaters
- Authors: Goetsch, Patricia-Ann
- Date: 1993
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:21060 , http://hdl.handle.net/10962/6190
- Description: 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.
- Full Text:
- Date Issued: 1993
- Authors: Goetsch, Patricia-Ann
- Date: 1993
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:21060 , http://hdl.handle.net/10962/6190
- Description: 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.
- Full Text:
- Date Issued: 1993
The culture of Dunaliella salina and the production of β-carotene in tannery effluents
- Authors: Laubscher, Richard Keith
- Date: 1992
- Subjects: Dunaliella , Carotenes , Tanneries -- Waste disposal , Recycling (Waste, etc.)
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4055 , http://hdl.handle.net/10962/d1004116 , Dunaliella , Carotenes , Tanneries -- Waste disposal , Recycling (Waste, etc.)
- Description: The problems of waste disposal in the tanning industry are unique in that the effluents are highly saline, have a high organic loading and contain heavy metals. Methods are available for the safe treatment and disposal of the latter two components, but the saline component requires the expensive outlay of evaporation ponds. This study has identified a possible use for the saline effluents, turning a problematic waste product into a potentially valuable by-product. A range of tannery effluents were identified and tested for their suitability for the mass cultivation of Dunaliella salina (bardawil strain). The bardawil strain was preferred over a local isolate because of its higher production of β-carotene. Ponded tannery effluents and combined processes effluent proved unsuitable for realistic propagation of the alga. Anaerobic digestion of combined processes effluent did not improve its suitability significantly. Anaerobic digestion of hide-soak effluent may remove persistent antimicrobial agents which influence algal growth, but its contribution to enhancing algal growth is equivocal. Undigested hide-soak effluent lacking in persistent antimicrobial agents was found to be an ideal culture medium, as no additional nutrients needed to be added. Significantly higher biomass was obtained in this effluent compared to chemically defined media. Induction of β-carotene was achieved in nitrogen-deficient defined media after culture in tannery effluent. This suggests that a two-stage system using hide-soak effluent for cell propagation and nitrogen deficient media for β-carotene induction, could be possible for the mass cultivation of D. salina for β-carotene production.
- Full Text:
- Date Issued: 1992
- Authors: Laubscher, Richard Keith
- Date: 1992
- Subjects: Dunaliella , Carotenes , Tanneries -- Waste disposal , Recycling (Waste, etc.)
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4055 , http://hdl.handle.net/10962/d1004116 , Dunaliella , Carotenes , Tanneries -- Waste disposal , Recycling (Waste, etc.)
- Description: The problems of waste disposal in the tanning industry are unique in that the effluents are highly saline, have a high organic loading and contain heavy metals. Methods are available for the safe treatment and disposal of the latter two components, but the saline component requires the expensive outlay of evaporation ponds. This study has identified a possible use for the saline effluents, turning a problematic waste product into a potentially valuable by-product. A range of tannery effluents were identified and tested for their suitability for the mass cultivation of Dunaliella salina (bardawil strain). The bardawil strain was preferred over a local isolate because of its higher production of β-carotene. Ponded tannery effluents and combined processes effluent proved unsuitable for realistic propagation of the alga. Anaerobic digestion of combined processes effluent did not improve its suitability significantly. Anaerobic digestion of hide-soak effluent may remove persistent antimicrobial agents which influence algal growth, but its contribution to enhancing algal growth is equivocal. Undigested hide-soak effluent lacking in persistent antimicrobial agents was found to be an ideal culture medium, as no additional nutrients needed to be added. Significantly higher biomass was obtained in this effluent compared to chemically defined media. Induction of β-carotene was achieved in nitrogen-deficient defined media after culture in tannery effluent. This suggests that a two-stage system using hide-soak effluent for cell propagation and nitrogen deficient media for β-carotene induction, could be possible for the mass cultivation of D. salina for β-carotene production.
- Full Text:
- Date Issued: 1992
The microbial production of polyphenol oxidase enzyme systems and their application in the treatment of phenolic wastewaters
- Scherman, Patricia Ann (neé Goetch)
- Authors: Scherman, Patricia Ann (neé Goetch)
- Date: 1992
- Subjects: Phenol oxidase Water -- Purification -- Biological treatment Enzymes -- Regulation
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4047 , http://hdl.handle.net/10962/d1004108
- Description: 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.
- Full Text:
- Date Issued: 1992
- Authors: Scherman, Patricia Ann (neé Goetch)
- Date: 1992
- Subjects: Phenol oxidase Water -- Purification -- Biological treatment Enzymes -- Regulation
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4047 , http://hdl.handle.net/10962/d1004108
- Description: 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.
- Full Text:
- Date Issued: 1992
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