Microalgal-bacterial flocs and extracellular polymeric substances for optimum function of integrated algal pond systems
- Authors: Jimoh, Taobat Adekilekun
- Date: 2021-10-29
- Subjects: Flocculation , Extracellular polymeric substances , Water Purification , Sewage Purification Anaerobic treatment , Integrated algae pond systems (IAPS) , Microalgal-bacterial flocs
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10962/191214 , vital:45071 , 10.21504/10962/191214
- Description: Despite the dire state of sanitation infrastructures, water scarcity, and the dwindling reserve of natural resources due to ever-increasing population growth, implementation of a suitable technology that can provide a solution to all these issues continues to be ignored. The integrated algal pond system (IAPS) is a wastewater treatment technology that combines the processes of anaerobic digestion and photosynthetic oxygenation to achieve wastewater treatment and facilitate the recovery of treated water and resources in the form of biogas and microalgal-bacterial biomass. The natural process of bioflocculation through microalgal-bacterial mutualism and production of extracellular polymeric substances (EPS) in high rate algal oxidation ponds (HRAOPs) of an IAPS increases efficiency of wastewater treatment and potentially enhances harvestability and biomass recovery, which could contribute significantly to the successful establishment of a biorefinery. Using a 500 PE pilot-scale IAPS supplied domestic sewage coupled with laboratory experiments, this study investigated the importance and function of in situ EPS production and MaB-floc formation in HRAOP. A metagenomic study revealed the biological components of the biomass or mixed liquor suspended solids (MLSS) produced in HRAOP and showed that the suspended biomass is composed largely of eukaryotes that were dominated by the colonial microalgae Pseudopediastrum sp. and Desmodesmus sp., and a diverse range of prokaryotes including bacteria and cyanobacteria. Dominance, within the bacterial population, by a sulphur-oxidizing bacterium, Thiothrix which comprised up to 80% of the prokaryotes, coincided with a period of poor flocculation and was therefore rationalized to have contributed to bulking and poor biomass settleability. Otherwise, good flocs were formed in the MLSS with settleability up to 95% and, within 1 h. The formation of MaB-flocs appeared to be dependent on EPS concentration of the mixed liquor due to the observed positive correlation between soluble EPS (S-EPS), biomass concentration, and settleability. The contribution and role of MLSS components towards the formation and sustenance of MaB-flocs were further demonstrated in laboratory experiments using pure strains of microalgae, cyanobacteria, and bacteria. Results showed that pure cultures of dominant microalgae in MLSS, Pseudopediastrum sp. and Desmodesmus sp. achieved a rapid 92 and 75% settleability within 3 h. A self-flocculating filamentous cyanobacterium, Leptolyngbya strain ECCN 20BG was isolated, characterized, and shown to achieve 99% settleability within 5 min by forming large tightly aggregated flocs. In further experiments, this strain was found to improve the settleability of MLSS by an average of 20%. Bacterial strains identified as Bacillus strain ECCN 40b, Bacillus strain ECCN 41b, Planococcus strain ECCN 45b, and Exiguobacterium strain ECCN 46b were also observed to produce sticky EPS-like materials in pure cultures that could also contribute to the aggregation of cells in a mixed environment. Given these results, various factors and/or mechanisms that might enhance microbial aggregation and biomass recovery from HRAOP MLSS were identified in this study and include; (1) dominance by larger colonial microalgae prevents disintegration of MaB-flocs and enhances recovery of biomass from MLSS by gravity sedimentation, (2) presence of filamentous cyanobacteria species that can self-flocculate to form an interwoven network of filaments may play an important role in the structural stability and settleability of MaB-flocs in MLSS, and (3) production of EPS to form the matrix or scaffold whereon all microbial components aggregate to develop a microenvironment. Indeed, all forms of EPS, except for that produced by Bacillus strain ECCN 41b, showed bioflocculating property and were able to serve as flocculants for the recovery of Chlorella, an alga known for its poor settleability. A combination of biochemical analyses and FTIR spectroscopy revealed the importance of carbohydrate enrichment of these biopolymers. Carbohydrate concentration in all forms of EPS was between 12 and 41% suggesting that production of these compounds by microbes within the MLSS contributed to MaB-floc formation. EPS extracted from bulk MLSS and EPS produced by Bacillus strains possessed some surface-active properties that were comparable to Triton X-100, indicating potential application in bioremediation and recovery of oil from contaminated soil and water. In particular, EPS generated from Bacillus strain ECCN 41b displayed relatively distinct properties including the quantity produced (> 500 mg/L), increased viscosity, inability to flocculate microalgal cells, a rhamnolipid content of 32%, and a higher surface-activity. Based on these results, Bacillus strain ECCN 41b was rationalized to produce anionic EPS with potential application in metal or oil recovery. In addition to EPS production, the bacteria Planococcus strain ECCN 45b and Exiguobacterium strain ECCN 46b appeared pigmented. Based on partial characterization using UV/Vis spectrophotometry, thin-layer chromatography, FTIR, and NMR, the pigments produced by these two strains appeared to be identical and were tentatively identified as ketocarotenoids. This study successfully demonstrated the importance of EPS production and formation of MaB-flocs in the MLSS from HRAOP of an IAPS treating domestic sewage. It is evident that increased settleability of the biomass does contribute to the reported efficiency of wastewater treatment by IAPS and would reduce both total suspended solids (TSS) and chemical oxygen demand (COD). In addition, demonstration that this biomass contains products of value such as carotenoids and EPS with potential for commercial use strengthens the idea of using IAPS as a platform technology for innovation of the wastewater treatment process to a biorefinery. , Thesis (PhD) -- Faculty of Science, Institute for Environmental Biotechnology, 2021
- Full Text:
- Date Issued: 2021-10-29
- Authors: Jimoh, Taobat Adekilekun
- Date: 2021-10-29
- Subjects: Flocculation , Extracellular polymeric substances , Water Purification , Sewage Purification Anaerobic treatment , Integrated algae pond systems (IAPS) , Microalgal-bacterial flocs
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10962/191214 , vital:45071 , 10.21504/10962/191214
- Description: Despite the dire state of sanitation infrastructures, water scarcity, and the dwindling reserve of natural resources due to ever-increasing population growth, implementation of a suitable technology that can provide a solution to all these issues continues to be ignored. The integrated algal pond system (IAPS) is a wastewater treatment technology that combines the processes of anaerobic digestion and photosynthetic oxygenation to achieve wastewater treatment and facilitate the recovery of treated water and resources in the form of biogas and microalgal-bacterial biomass. The natural process of bioflocculation through microalgal-bacterial mutualism and production of extracellular polymeric substances (EPS) in high rate algal oxidation ponds (HRAOPs) of an IAPS increases efficiency of wastewater treatment and potentially enhances harvestability and biomass recovery, which could contribute significantly to the successful establishment of a biorefinery. Using a 500 PE pilot-scale IAPS supplied domestic sewage coupled with laboratory experiments, this study investigated the importance and function of in situ EPS production and MaB-floc formation in HRAOP. A metagenomic study revealed the biological components of the biomass or mixed liquor suspended solids (MLSS) produced in HRAOP and showed that the suspended biomass is composed largely of eukaryotes that were dominated by the colonial microalgae Pseudopediastrum sp. and Desmodesmus sp., and a diverse range of prokaryotes including bacteria and cyanobacteria. Dominance, within the bacterial population, by a sulphur-oxidizing bacterium, Thiothrix which comprised up to 80% of the prokaryotes, coincided with a period of poor flocculation and was therefore rationalized to have contributed to bulking and poor biomass settleability. Otherwise, good flocs were formed in the MLSS with settleability up to 95% and, within 1 h. The formation of MaB-flocs appeared to be dependent on EPS concentration of the mixed liquor due to the observed positive correlation between soluble EPS (S-EPS), biomass concentration, and settleability. The contribution and role of MLSS components towards the formation and sustenance of MaB-flocs were further demonstrated in laboratory experiments using pure strains of microalgae, cyanobacteria, and bacteria. Results showed that pure cultures of dominant microalgae in MLSS, Pseudopediastrum sp. and Desmodesmus sp. achieved a rapid 92 and 75% settleability within 3 h. A self-flocculating filamentous cyanobacterium, Leptolyngbya strain ECCN 20BG was isolated, characterized, and shown to achieve 99% settleability within 5 min by forming large tightly aggregated flocs. In further experiments, this strain was found to improve the settleability of MLSS by an average of 20%. Bacterial strains identified as Bacillus strain ECCN 40b, Bacillus strain ECCN 41b, Planococcus strain ECCN 45b, and Exiguobacterium strain ECCN 46b were also observed to produce sticky EPS-like materials in pure cultures that could also contribute to the aggregation of cells in a mixed environment. Given these results, various factors and/or mechanisms that might enhance microbial aggregation and biomass recovery from HRAOP MLSS were identified in this study and include; (1) dominance by larger colonial microalgae prevents disintegration of MaB-flocs and enhances recovery of biomass from MLSS by gravity sedimentation, (2) presence of filamentous cyanobacteria species that can self-flocculate to form an interwoven network of filaments may play an important role in the structural stability and settleability of MaB-flocs in MLSS, and (3) production of EPS to form the matrix or scaffold whereon all microbial components aggregate to develop a microenvironment. Indeed, all forms of EPS, except for that produced by Bacillus strain ECCN 41b, showed bioflocculating property and were able to serve as flocculants for the recovery of Chlorella, an alga known for its poor settleability. A combination of biochemical analyses and FTIR spectroscopy revealed the importance of carbohydrate enrichment of these biopolymers. Carbohydrate concentration in all forms of EPS was between 12 and 41% suggesting that production of these compounds by microbes within the MLSS contributed to MaB-floc formation. EPS extracted from bulk MLSS and EPS produced by Bacillus strains possessed some surface-active properties that were comparable to Triton X-100, indicating potential application in bioremediation and recovery of oil from contaminated soil and water. In particular, EPS generated from Bacillus strain ECCN 41b displayed relatively distinct properties including the quantity produced (> 500 mg/L), increased viscosity, inability to flocculate microalgal cells, a rhamnolipid content of 32%, and a higher surface-activity. Based on these results, Bacillus strain ECCN 41b was rationalized to produce anionic EPS with potential application in metal or oil recovery. In addition to EPS production, the bacteria Planococcus strain ECCN 45b and Exiguobacterium strain ECCN 46b appeared pigmented. Based on partial characterization using UV/Vis spectrophotometry, thin-layer chromatography, FTIR, and NMR, the pigments produced by these two strains appeared to be identical and were tentatively identified as ketocarotenoids. This study successfully demonstrated the importance of EPS production and formation of MaB-flocs in the MLSS from HRAOP of an IAPS treating domestic sewage. It is evident that increased settleability of the biomass does contribute to the reported efficiency of wastewater treatment by IAPS and would reduce both total suspended solids (TSS) and chemical oxygen demand (COD). In addition, demonstration that this biomass contains products of value such as carotenoids and EPS with potential for commercial use strengthens the idea of using IAPS as a platform technology for innovation of the wastewater treatment process to a biorefinery. , Thesis (PhD) -- Faculty of Science, Institute for Environmental Biotechnology, 2021
- Full Text:
- Date Issued: 2021-10-29
Mitigating salt accumulation in recycled brewery effluent through the integration of water treatment, agriculture and aquaculture
- Authors: Mabasa, Nyiko Charity
- Date: 2021-10-29
- Subjects: Brewery waste South Africa Eastern Cape , Recycling (Waste, etc.) South Africa Eastern Cape , Water reuse South Africa Eastern Cape , Irrigation South Africa Eastern Cape , Sewage Purification Anaerobic treatment , Constructed wetlands , Aquaculture
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10962/191126 , vital:45063 , 10.21504/10962/191126
- Description: Water scarcity in South Africa, and globally, presents challenges for industries. It is imperative to develop responsible water use, such as recycling and reusing wastewater from food processing industries such as breweries. The Ibhayi Brewery (SAB Ltd) employs a combination of sustainable treatment processes that include anaerobic digestion (AD), primary facultative ponds (PFP), high rate algal ponds (HRAP) and constructed wetlands (CW) to treat brewery effluent on an experimental scale. The constituent concentrations of these experimentally treated effluents are within the ranges prescribed by local regulations to allow for potential downstream use in agriculture and aquaculture. However, the sodium content in this treated effluent, which originates from upstream cleaning agents and pH control at the onsite effluent treatment facility, is a constraint to the downstream use of brewery effluent. This study addresses the salt problem, by investigating the potential of either reducing/eliminating salt addition at source, or developing alternative techniques for downstream agriculture to mitigate the effects of salt accumulation caused by irrigation with brewery effluent. Four salt-tolerant test crops; Swiss chard (Beta vulgaris), saltbush (Atriplex nummularia), Salicornia meyeriana and sorghum (Sorghum bicolor), grew efficiently in brewery effluent irrigated soils but did not stop sodium accumulation in the growth medium. Swiss chard had the best growth with a wet biomass accumulation of 8,173 g m-2, due to the plant’s ability to tolerate saline conditions and continuous cropping. Crop rotation, to limit effects of nutrient depletion in soil, had no significant effect on plant growth suggesting soils were adequately able to provide micro-nutrients in the short-term. Prolonged irrigation with brewery effluent can lead to sodium accumulation in the soil, which was successfully controlled through the addition of soil amendments (gypsum and Trichoderma cultures). These reduced soil sodium from a potentially limiting level of 1,398 mg L-1 to the acceptable levels of 240 mg L-1 and 353 mg L-1 respectively, mainly through leaching. However only Trichoderma improved Swiss chard production to 11,238 g m-2. While crop rotation in this work did not contribute to mitigating the problem of salt accumulation, soil amended with Trichoderma appears to be a potential solution when brewery effluent is reused in agriculture. In an alternative to soil cultivation, CWs were trialled with no significant differences in the sodium concentration of brewery effluent treated along a 15 m lateral flow CW, which could be attributed to evapotranspiration. This was notably accompanied by a desirable 95.21% decrease in ammonia from inlet to outlet resulting in significant improvement in water quality for reuse in aquaculture where ammonia levels are important limiting constraints. While CWs remain a suitable brewery effluent treatment solution, this technology requires additional modelling and optimisation in order to mitigate the problem of salt accumulation in the reuse of treated brewery effluent in agriculture and aquaculture. This research demonstrates the baseline information for such modelling and optimisation. African catfish (Clarias gariepinus) grew in CW treated brewery effluent; however, this growth was moderate at 0.92% bw day-1, whereas Mozambique tilapia (Oreochromis mossambicus) were shown to be unsuited to growth in this system and lost weight with an average specific growth rate (SGR) of -0.98% bw day-1; and both fish species presenting with health related concerns. Hardy fish species such as African catfish can be cultured in brewery effluent, but with risk involved. This was a preliminary study to develop parameters for future dimensional analysis modelling to allow optimisation of the CW, based on nutrient removal rates obtained which will allow for improved downstream aquaculture by reducing or eliminating risks presented in this study. This work has also contributed to a foundation for the development of guidelines that use a risk-based approach for water use in aquaculture. Alternatives to the current in place cleaning agents were considered to mitigate the effects of salt accumulation. Sodium is introduced into the effluent via the use of sodium hydroxide and sodium chlorite for cleaning and disinfection in the brewery, as well as through effluent pH adjustment in the AD plant. The widespread use of outdated legacy cleaning systems and pH adjustment regimes is entrenched in the brewery standard operating procedures (SOP). A cost-benefit analysis (CBA) demonstrated that a change of cleaning and disinfecting regimes to hydrogen peroxide in the brewery, and magnesium hydroxide pH adjustment in the effluent treatment plant addresses the sodium issue upstream in the brewery practically eliminating sodium from the effluent. In addition, a life cycle analysis (LCA) was carried out to assess the environmental impacts associated with the alternative cleaning and pH adjustment scenarios. The LCA showed that electricity consumption during use phase of the chemicals for respective purposes, as well as their production activities were major contributors to the significant environmental impact categories that were assessed. The cleaning scenario employing the use of hydrogen peroxide for both cleaning and disinfection was found to be the most environmentally sustainable. This was attributed to the reduced number of chemicals used compared to the other cleaning scenarios. Dolomitic lime was the pH adjustment alternative with the lowest average environmental impact; but, however, had a higher impact on freshwater eutrophication which is of major concern if the effluent will be reused for irrigation. Magnesium hydroxide was therefore considered to be the better option as a sodium hydroxide alternative for pH adjustment. This mitigates salt accumulation, making treated brewery effluent suitable for reuse in high value downstream agriculture and aquaculture, while employing more environmentally sustainable technologies. Notably, this converts brewery effluent from a financial liability to Ibhayi Brewery, into a product containing water and nutrients that generate income, improve food security, and can create employment in downstream agriculture and aquaculture in a sustainable manner. , Thesis (PhD) -- Faculty of Science, Ichthyology and Fisheries Science, 2021
- Full Text:
- Date Issued: 2021-10-29
- Authors: Mabasa, Nyiko Charity
- Date: 2021-10-29
- Subjects: Brewery waste South Africa Eastern Cape , Recycling (Waste, etc.) South Africa Eastern Cape , Water reuse South Africa Eastern Cape , Irrigation South Africa Eastern Cape , Sewage Purification Anaerobic treatment , Constructed wetlands , Aquaculture
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10962/191126 , vital:45063 , 10.21504/10962/191126
- Description: Water scarcity in South Africa, and globally, presents challenges for industries. It is imperative to develop responsible water use, such as recycling and reusing wastewater from food processing industries such as breweries. The Ibhayi Brewery (SAB Ltd) employs a combination of sustainable treatment processes that include anaerobic digestion (AD), primary facultative ponds (PFP), high rate algal ponds (HRAP) and constructed wetlands (CW) to treat brewery effluent on an experimental scale. The constituent concentrations of these experimentally treated effluents are within the ranges prescribed by local regulations to allow for potential downstream use in agriculture and aquaculture. However, the sodium content in this treated effluent, which originates from upstream cleaning agents and pH control at the onsite effluent treatment facility, is a constraint to the downstream use of brewery effluent. This study addresses the salt problem, by investigating the potential of either reducing/eliminating salt addition at source, or developing alternative techniques for downstream agriculture to mitigate the effects of salt accumulation caused by irrigation with brewery effluent. Four salt-tolerant test crops; Swiss chard (Beta vulgaris), saltbush (Atriplex nummularia), Salicornia meyeriana and sorghum (Sorghum bicolor), grew efficiently in brewery effluent irrigated soils but did not stop sodium accumulation in the growth medium. Swiss chard had the best growth with a wet biomass accumulation of 8,173 g m-2, due to the plant’s ability to tolerate saline conditions and continuous cropping. Crop rotation, to limit effects of nutrient depletion in soil, had no significant effect on plant growth suggesting soils were adequately able to provide micro-nutrients in the short-term. Prolonged irrigation with brewery effluent can lead to sodium accumulation in the soil, which was successfully controlled through the addition of soil amendments (gypsum and Trichoderma cultures). These reduced soil sodium from a potentially limiting level of 1,398 mg L-1 to the acceptable levels of 240 mg L-1 and 353 mg L-1 respectively, mainly through leaching. However only Trichoderma improved Swiss chard production to 11,238 g m-2. While crop rotation in this work did not contribute to mitigating the problem of salt accumulation, soil amended with Trichoderma appears to be a potential solution when brewery effluent is reused in agriculture. In an alternative to soil cultivation, CWs were trialled with no significant differences in the sodium concentration of brewery effluent treated along a 15 m lateral flow CW, which could be attributed to evapotranspiration. This was notably accompanied by a desirable 95.21% decrease in ammonia from inlet to outlet resulting in significant improvement in water quality for reuse in aquaculture where ammonia levels are important limiting constraints. While CWs remain a suitable brewery effluent treatment solution, this technology requires additional modelling and optimisation in order to mitigate the problem of salt accumulation in the reuse of treated brewery effluent in agriculture and aquaculture. This research demonstrates the baseline information for such modelling and optimisation. African catfish (Clarias gariepinus) grew in CW treated brewery effluent; however, this growth was moderate at 0.92% bw day-1, whereas Mozambique tilapia (Oreochromis mossambicus) were shown to be unsuited to growth in this system and lost weight with an average specific growth rate (SGR) of -0.98% bw day-1; and both fish species presenting with health related concerns. Hardy fish species such as African catfish can be cultured in brewery effluent, but with risk involved. This was a preliminary study to develop parameters for future dimensional analysis modelling to allow optimisation of the CW, based on nutrient removal rates obtained which will allow for improved downstream aquaculture by reducing or eliminating risks presented in this study. This work has also contributed to a foundation for the development of guidelines that use a risk-based approach for water use in aquaculture. Alternatives to the current in place cleaning agents were considered to mitigate the effects of salt accumulation. Sodium is introduced into the effluent via the use of sodium hydroxide and sodium chlorite for cleaning and disinfection in the brewery, as well as through effluent pH adjustment in the AD plant. The widespread use of outdated legacy cleaning systems and pH adjustment regimes is entrenched in the brewery standard operating procedures (SOP). A cost-benefit analysis (CBA) demonstrated that a change of cleaning and disinfecting regimes to hydrogen peroxide in the brewery, and magnesium hydroxide pH adjustment in the effluent treatment plant addresses the sodium issue upstream in the brewery practically eliminating sodium from the effluent. In addition, a life cycle analysis (LCA) was carried out to assess the environmental impacts associated with the alternative cleaning and pH adjustment scenarios. The LCA showed that electricity consumption during use phase of the chemicals for respective purposes, as well as their production activities were major contributors to the significant environmental impact categories that were assessed. The cleaning scenario employing the use of hydrogen peroxide for both cleaning and disinfection was found to be the most environmentally sustainable. This was attributed to the reduced number of chemicals used compared to the other cleaning scenarios. Dolomitic lime was the pH adjustment alternative with the lowest average environmental impact; but, however, had a higher impact on freshwater eutrophication which is of major concern if the effluent will be reused for irrigation. Magnesium hydroxide was therefore considered to be the better option as a sodium hydroxide alternative for pH adjustment. This mitigates salt accumulation, making treated brewery effluent suitable for reuse in high value downstream agriculture and aquaculture, while employing more environmentally sustainable technologies. Notably, this converts brewery effluent from a financial liability to Ibhayi Brewery, into a product containing water and nutrients that generate income, improve food security, and can create employment in downstream agriculture and aquaculture in a sustainable manner. , Thesis (PhD) -- Faculty of Science, Ichthyology and Fisheries Science, 2021
- Full Text:
- Date Issued: 2021-10-29
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