Performance of an integrated algal pond for treatment of domestic sewage: a process audit
- Authors: Dube, Anele
- Date: 2020
- Subjects: Water -- Purification , Sewage -- Purification -- Anaerobic treatment , Algae -- Biotechnology , Waste disposal -- South Africa , Integrated algae pond systems (IAPS)
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/167043 , vital:41432
- Description: Integrated algae pond systems (IAPS) are energy efficient, robust, passive systems that use the principles of fermentation, photosynthesis and microbial metabolism to remediate wastewater, producing a good quality effluent with reuse potential. In addition to the treatment of wastewater, IAPS have the ability to generate two additional product streams viz. biogas and biomass. The latter adds to the attractiveness of the system. However, the implementation of this technology, like many passive systems, has remained limited at a commercial scale, and the inclination is still towards grey technologies. The aim of this research was to investigate the capabilities and potential of a demonstration-scale IAPS and use results obtained to establish a process audit framework. The aspects considered for the audit included performance efficiency, effluent water quality, biomass composition, quantity and productivity within the ponds, and cost analysis of operation and maintenance over a 9-year period. Plant performance was closely monitored during the course of the study and this led to a review of previously adopted plant management strategies. Troubleshooting exercises were also carried out when plant performance declined. Results showed that IAPS efficiently reduced standard water parameters with the exception of pH, dissolved oxygen, and nitrate whose values increased from raw influent to final effluent. The following water quality parameters were established for the final effluent: total suspended solids 55 ± 7.1 mg. L-1 (n = 28); chemical oxygen demand 94.1 ± 10.6 mg. L-1 (n = 28) (after removal of algae); pH 9.9 ± 0.01 (n = 26); ammonium nitrogen 1.7 ± 0.3 mg. L-1 (n = 25); nitrate 3.3 ± 0.6 mg. L-1 (n = 25); ortho-phosphate 1.6 ± 0.2 mg. L-1 (n = 25); electrical conductivity 98.7 ± 2.0 mS m-1 (n = 26) and faecal coliforms (per 100 mL) 1482.6 ± 636.0 (n = 24). The final effluent measured consistently high chemical oxygen demand and total suspended solids, however close analysis showed that total suspended solids could be controlled by increasing the frequency of removal of settled biomass within the settling ponds. Biomass produced contained microalgae, bacteria, metazoa, and protozoa. The biomass productivity achieved was as high as 130.6 kg ha-1 d-1; however, about 33% was lost to the final effluent due to inadequate settling. Results obtained during the course of this study and outcomes of earlier work on IAPS are taken as the baseline to determine parameters needed for the development of the process audit framework. Techniques utilised to derive the blue print process audit protocol for IAPS included a turtle diagram, a flow diagram and a checklist. Attention to plant management proved vital in determining overall performance. Cost, including operating and maintenance, of treating water using the demonstration scale system on a per person equivalent per year basis was determined as ZAR 123.87 (where, ZAR to USD = 0.07).
- Full Text:
- Date Issued: 2020
- Authors: Dube, Anele
- Date: 2020
- Subjects: Water -- Purification , Sewage -- Purification -- Anaerobic treatment , Algae -- Biotechnology , Waste disposal -- South Africa , Integrated algae pond systems (IAPS)
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/167043 , vital:41432
- Description: Integrated algae pond systems (IAPS) are energy efficient, robust, passive systems that use the principles of fermentation, photosynthesis and microbial metabolism to remediate wastewater, producing a good quality effluent with reuse potential. In addition to the treatment of wastewater, IAPS have the ability to generate two additional product streams viz. biogas and biomass. The latter adds to the attractiveness of the system. However, the implementation of this technology, like many passive systems, has remained limited at a commercial scale, and the inclination is still towards grey technologies. The aim of this research was to investigate the capabilities and potential of a demonstration-scale IAPS and use results obtained to establish a process audit framework. The aspects considered for the audit included performance efficiency, effluent water quality, biomass composition, quantity and productivity within the ponds, and cost analysis of operation and maintenance over a 9-year period. Plant performance was closely monitored during the course of the study and this led to a review of previously adopted plant management strategies. Troubleshooting exercises were also carried out when plant performance declined. Results showed that IAPS efficiently reduced standard water parameters with the exception of pH, dissolved oxygen, and nitrate whose values increased from raw influent to final effluent. The following water quality parameters were established for the final effluent: total suspended solids 55 ± 7.1 mg. L-1 (n = 28); chemical oxygen demand 94.1 ± 10.6 mg. L-1 (n = 28) (after removal of algae); pH 9.9 ± 0.01 (n = 26); ammonium nitrogen 1.7 ± 0.3 mg. L-1 (n = 25); nitrate 3.3 ± 0.6 mg. L-1 (n = 25); ortho-phosphate 1.6 ± 0.2 mg. L-1 (n = 25); electrical conductivity 98.7 ± 2.0 mS m-1 (n = 26) and faecal coliforms (per 100 mL) 1482.6 ± 636.0 (n = 24). The final effluent measured consistently high chemical oxygen demand and total suspended solids, however close analysis showed that total suspended solids could be controlled by increasing the frequency of removal of settled biomass within the settling ponds. Biomass produced contained microalgae, bacteria, metazoa, and protozoa. The biomass productivity achieved was as high as 130.6 kg ha-1 d-1; however, about 33% was lost to the final effluent due to inadequate settling. Results obtained during the course of this study and outcomes of earlier work on IAPS are taken as the baseline to determine parameters needed for the development of the process audit framework. Techniques utilised to derive the blue print process audit protocol for IAPS included a turtle diagram, a flow diagram and a checklist. Attention to plant management proved vital in determining overall performance. Cost, including operating and maintenance, of treating water using the demonstration scale system on a per person equivalent per year basis was determined as ZAR 123.87 (where, ZAR to USD = 0.07).
- Full Text:
- Date Issued: 2020
Post-treatment technologies for integrated algal pond systems
- Authors: Westensee, Dirk Karl
- Date: 2015
- Subjects: Sewage disposal plants -- South Africa -- Grahamstown , Water -- Purification -- Filtration , Water -- Purification -- Slow sand filtration , Sewage -- Purification -- Anaerobic treatment , Sewage -- Purification -- Biological treatment , Algae -- Biotechnology
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5958 , http://hdl.handle.net/10962/d1018180
- Description: Integrated Algae Pond Systems (IAPS) are a derivation of the Oswald designed Algal Integrated Wastewater Pond Systems (AIWPS®) and combine the use of anaerobic and aerobic bioprocesses to effect wastewater treatment. IAPS technology was introduced to South Africa in 1996 and a pilot plant designed and commissioned at the Belmont Valley WWTW in Grahamstown. The system has been in continual use since implementation and affords a secondarily treated water for reclamation according to its design specifications which most closely resemble those of the AIWPS® Advanced Secondary Process developed by Oswald. As a consequence, and as might be expected, while the technology performed well and delivered a final effluent superior to most pond systems deployed in South Africa it was unable to meet The Department of Water Affairs General Standard for nutrient removal and effluent discharge. The work described in this thesis involved the design, construction, and evaluation of several tertiary treatment units (TTU') for incorporation into the IAPS process design. Included were; Maturation Ponds (MP), Slow Sand Filter (SSF) and Rock Filters (RF). Three MP's were constructed in series with a 12 day retention time and operated in parallel with a two-layered SSF and a three-stage RF. Water quality of the effluent emerging from each of these TTU's was monitored over a 10 month period. Significant decreases in the chemical oxygen demand (COD), ammonium-N, phosphate-P, nitrate-N, faecal coliforms (FC) and total coliforms (TC) were achieved by these TTU's. On average, throughout the testing period, water quality was within the statutory limit for discharge to a water course that is not a listed water course, with the exception of the total suspended solids (TSS). The RF was determined as the most suitable TTU for commercial use due to production of a better quality water, smaller footprint, lower construction costs and less maintenance required. From the results of this investigation it is concluded that commercial deployment of IAPS for the treatment of municipal sewage requires the inclusion of a suitable TTU. Furthermore, and based on the findings presented, RF appears most appropriate to ensure that quality of the final effluent meets the standard for discharge.
- Full Text:
- Date Issued: 2015
- Authors: Westensee, Dirk Karl
- Date: 2015
- Subjects: Sewage disposal plants -- South Africa -- Grahamstown , Water -- Purification -- Filtration , Water -- Purification -- Slow sand filtration , Sewage -- Purification -- Anaerobic treatment , Sewage -- Purification -- Biological treatment , Algae -- Biotechnology
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5958 , http://hdl.handle.net/10962/d1018180
- Description: Integrated Algae Pond Systems (IAPS) are a derivation of the Oswald designed Algal Integrated Wastewater Pond Systems (AIWPS®) and combine the use of anaerobic and aerobic bioprocesses to effect wastewater treatment. IAPS technology was introduced to South Africa in 1996 and a pilot plant designed and commissioned at the Belmont Valley WWTW in Grahamstown. The system has been in continual use since implementation and affords a secondarily treated water for reclamation according to its design specifications which most closely resemble those of the AIWPS® Advanced Secondary Process developed by Oswald. As a consequence, and as might be expected, while the technology performed well and delivered a final effluent superior to most pond systems deployed in South Africa it was unable to meet The Department of Water Affairs General Standard for nutrient removal and effluent discharge. The work described in this thesis involved the design, construction, and evaluation of several tertiary treatment units (TTU') for incorporation into the IAPS process design. Included were; Maturation Ponds (MP), Slow Sand Filter (SSF) and Rock Filters (RF). Three MP's were constructed in series with a 12 day retention time and operated in parallel with a two-layered SSF and a three-stage RF. Water quality of the effluent emerging from each of these TTU's was monitored over a 10 month period. Significant decreases in the chemical oxygen demand (COD), ammonium-N, phosphate-P, nitrate-N, faecal coliforms (FC) and total coliforms (TC) were achieved by these TTU's. On average, throughout the testing period, water quality was within the statutory limit for discharge to a water course that is not a listed water course, with the exception of the total suspended solids (TSS). The RF was determined as the most suitable TTU for commercial use due to production of a better quality water, smaller footprint, lower construction costs and less maintenance required. From the results of this investigation it is concluded that commercial deployment of IAPS for the treatment of municipal sewage requires the inclusion of a suitable TTU. Furthermore, and based on the findings presented, RF appears most appropriate to ensure that quality of the final effluent meets the standard for discharge.
- Full Text:
- Date Issued: 2015
Exploring the fertiliser potential of biosolids from algae integrated wastewater treatment systems
- Authors: Mlambo, Patricia Zanele
- Date: 2014
- Subjects: Sewage disposal plants , Sewage sludge as fertilizer , Algae -- Biotechnology , Sewage -- Purification -- Anaerobic treatment , Plant regulators , Biofertilizers , Microalgae -- Biotechnology
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5957 , http://hdl.handle.net/10962/d1013342
- Description: High rate algae oxidation ponds (HRAOP) for domestic wastewater treatment generate biosolids that are predominantly microalgae. Consequently, HRAOP biosolids are enriched with minerals, amino acids, nutrients and possibly contain plant growth regulator (PGR)-like substances, which makes HRAOP biosolids attractive as fertiliser or PGR. This study investigated HRAOP biosolids as a starting material for a natural, cost-effective and readily-available eco-friendly organic fertiliser and/or PGRs. Various HRAOP extract formulations were prepared and their effect on plant growth and development was evaluated using selected bioassays. Initial screening included assessing the effect on change in specific leaf area, radish cotyledon expansion as an indicator of PGR-like activity, and seed germination index (GI). More detailed studies on fertiliser efficacy and PGR-like activity utilised bean (Phaseolus vulgaris) and tomato (Solanum lycopersicum) plants. Combined effects of sonicated (S) and 40% v/v methanol (M) extract (5:1 SM) had impressive plant responses, comparable to Hoagland solution (HS). Other potentially fertiliser formulations included 0.5% M, 1% M, 2.5% S and 5% S formulations. The 5:1 SM and 5% S showed greater PGR-like activity, promoting cotyledon expansion by 459 ± 0.02% and 362 ± 0.01%, respectively. GI data showed that none of the formulations negatively impacted germination. Further investigation showed that the 5% S formulation increased leaf length, width and area by 6.69 ± 0.24, 6.21 ± 0.2 mm and 41.55 ± 0.2 mm². All formulated fertiliser extracts had no adverse effect on chlorophyll content and plant nutrient balance as indicated by C:N (8-10:1) ratio. In addition, plants appeared to actively mobilise nutrients to regions where needed as evidenced by a shift in shoot: root ratio depending on C, N and water availability. Furthermore, 5% S caused a 75% increase in tomato productivity and had no effect on bean productivity. Whereas, 5:1 SM and 1% M formulation improved bean pod production by 33.3% and 11%, respectively but did not affect tomato production. Harvest index (HI) however indicated a 3% reduction in tomato productivity with 5:1 SM and little or no enhancement in bean productivity with both 5:1 SM and 5% S treatments. Bean plants treated with 5:1 SM and 5% S produced larger fruits, which could be an indication of the presence of a PGR effect. Overall, HRAOP biosolids extracts prepared and investigated in this study demonstrated both fertiliser characteristics and PGR-like activity with performances comparable and in some cases exceeding that of commercial products. However additional research is needed to confirm presence of PGR-like activities and fertiliser efficacy.
- Full Text:
- Date Issued: 2014
- Authors: Mlambo, Patricia Zanele
- Date: 2014
- Subjects: Sewage disposal plants , Sewage sludge as fertilizer , Algae -- Biotechnology , Sewage -- Purification -- Anaerobic treatment , Plant regulators , Biofertilizers , Microalgae -- Biotechnology
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5957 , http://hdl.handle.net/10962/d1013342
- Description: High rate algae oxidation ponds (HRAOP) for domestic wastewater treatment generate biosolids that are predominantly microalgae. Consequently, HRAOP biosolids are enriched with minerals, amino acids, nutrients and possibly contain plant growth regulator (PGR)-like substances, which makes HRAOP biosolids attractive as fertiliser or PGR. This study investigated HRAOP biosolids as a starting material for a natural, cost-effective and readily-available eco-friendly organic fertiliser and/or PGRs. Various HRAOP extract formulations were prepared and their effect on plant growth and development was evaluated using selected bioassays. Initial screening included assessing the effect on change in specific leaf area, radish cotyledon expansion as an indicator of PGR-like activity, and seed germination index (GI). More detailed studies on fertiliser efficacy and PGR-like activity utilised bean (Phaseolus vulgaris) and tomato (Solanum lycopersicum) plants. Combined effects of sonicated (S) and 40% v/v methanol (M) extract (5:1 SM) had impressive plant responses, comparable to Hoagland solution (HS). Other potentially fertiliser formulations included 0.5% M, 1% M, 2.5% S and 5% S formulations. The 5:1 SM and 5% S showed greater PGR-like activity, promoting cotyledon expansion by 459 ± 0.02% and 362 ± 0.01%, respectively. GI data showed that none of the formulations negatively impacted germination. Further investigation showed that the 5% S formulation increased leaf length, width and area by 6.69 ± 0.24, 6.21 ± 0.2 mm and 41.55 ± 0.2 mm². All formulated fertiliser extracts had no adverse effect on chlorophyll content and plant nutrient balance as indicated by C:N (8-10:1) ratio. In addition, plants appeared to actively mobilise nutrients to regions where needed as evidenced by a shift in shoot: root ratio depending on C, N and water availability. Furthermore, 5% S caused a 75% increase in tomato productivity and had no effect on bean productivity. Whereas, 5:1 SM and 1% M formulation improved bean pod production by 33.3% and 11%, respectively but did not affect tomato production. Harvest index (HI) however indicated a 3% reduction in tomato productivity with 5:1 SM and little or no enhancement in bean productivity with both 5:1 SM and 5% S treatments. Bean plants treated with 5:1 SM and 5% S produced larger fruits, which could be an indication of the presence of a PGR effect. Overall, HRAOP biosolids extracts prepared and investigated in this study demonstrated both fertiliser characteristics and PGR-like activity with performances comparable and in some cases exceeding that of commercial products. However additional research is needed to confirm presence of PGR-like activities and fertiliser efficacy.
- Full Text:
- Date Issued: 2014
The water and nutrient potential of brewery effluent for hydroponic tomato production
- Authors: Power, Sean Duncan
- Date: 2014
- Subjects: Hydroponics , Tomatoes -- Breeding , Brewery waste , Water -- Purification , Algae culture , Algae -- Biotechnology , Nitric acid , Phosphoric acid
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5358 , http://hdl.handle.net/10962/d1011604 , Hydroponics , Tomatoes -- Breeding , Brewery waste , Water -- Purification , Algae culture , Algae -- Biotechnology , Nitric acid , Phosphoric acid
- Description: Brewery effluent that had undergone treatment in an anaerobic digester (AD) was used as an alternative water and nutrient source for hydroponic crop production. Brewery effluent was demonstrated to contain sufficient nutrients to support the growth, flowering and fruiting of Lycopersicum escolentum "Moneymaker" tomato crops. The adjustment of the effluent pH with phosphoric acid to between pH 6.0 and 6.5 increased the development of the crops by around 100% compared to crops grown in unaltered effluent. The pH adjusted effluent-grown plants grew to a mean height of 831.4 ± 21.1 mm and a dry biomass weight of 42.34 ± 2.76 g compared to the unaltered pH effluent plants which grew to a height of 410.6 ± 20.5 mm and a weight of 7.65 ± 0.68 g after 49 days. Effluent treatment in high-rate algal ponds (HRAP) was determined to have no positive effect on the nutritional potential of the effluent for Moneymaker production. The effluent-grown plants did not perform as well as plants grown in inorganic-fertilizer and municipal water. Plants grown in effluent grew taller but did not produce significantly more fruit when phosphoric acid (height: 1573.3 ± 50.4 mm, 19.4 ± 1.4 fruit per plant) was compared to nitric acid (height: 1254.1 ± 25.4 mm, 15.6 ± 1.5 fruit per plant) as the pH adjustment over 72 days. Direct and secondary plant stresses from effluent alkalinity, ammonium nutrition, nitrogen limitation, sodium concentrations and heat stress among other factors were probably confounding variables in these trials and require further investigation. Considering the raw effluent composition and manipulating the AD operation is a potential opportunity to improve overall AD performance, reduce chemical inputs in the effluent treatment process, reduce the final effluent alkalinity, and increase available nitrogen content in the final effluent. The anaerobic digester discharging >1000 m³ of nutrient enriched effluent every day is a resource with considerable potential. The benefits of developing this resource can contribute to cost-reduction at the brewery, more efficient water, nutrient and energy management at the brewery, and offer opportunities for job creation and potentially benefit local food security.
- Full Text:
- Date Issued: 2014
- Authors: Power, Sean Duncan
- Date: 2014
- Subjects: Hydroponics , Tomatoes -- Breeding , Brewery waste , Water -- Purification , Algae culture , Algae -- Biotechnology , Nitric acid , Phosphoric acid
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5358 , http://hdl.handle.net/10962/d1011604 , Hydroponics , Tomatoes -- Breeding , Brewery waste , Water -- Purification , Algae culture , Algae -- Biotechnology , Nitric acid , Phosphoric acid
- Description: Brewery effluent that had undergone treatment in an anaerobic digester (AD) was used as an alternative water and nutrient source for hydroponic crop production. Brewery effluent was demonstrated to contain sufficient nutrients to support the growth, flowering and fruiting of Lycopersicum escolentum "Moneymaker" tomato crops. The adjustment of the effluent pH with phosphoric acid to between pH 6.0 and 6.5 increased the development of the crops by around 100% compared to crops grown in unaltered effluent. The pH adjusted effluent-grown plants grew to a mean height of 831.4 ± 21.1 mm and a dry biomass weight of 42.34 ± 2.76 g compared to the unaltered pH effluent plants which grew to a height of 410.6 ± 20.5 mm and a weight of 7.65 ± 0.68 g after 49 days. Effluent treatment in high-rate algal ponds (HRAP) was determined to have no positive effect on the nutritional potential of the effluent for Moneymaker production. The effluent-grown plants did not perform as well as plants grown in inorganic-fertilizer and municipal water. Plants grown in effluent grew taller but did not produce significantly more fruit when phosphoric acid (height: 1573.3 ± 50.4 mm, 19.4 ± 1.4 fruit per plant) was compared to nitric acid (height: 1254.1 ± 25.4 mm, 15.6 ± 1.5 fruit per plant) as the pH adjustment over 72 days. Direct and secondary plant stresses from effluent alkalinity, ammonium nutrition, nitrogen limitation, sodium concentrations and heat stress among other factors were probably confounding variables in these trials and require further investigation. Considering the raw effluent composition and manipulating the AD operation is a potential opportunity to improve overall AD performance, reduce chemical inputs in the effluent treatment process, reduce the final effluent alkalinity, and increase available nitrogen content in the final effluent. The anaerobic digester discharging >1000 m³ of nutrient enriched effluent every day is a resource with considerable potential. The benefits of developing this resource can contribute to cost-reduction at the brewery, more efficient water, nutrient and energy management at the brewery, and offer opportunities for job creation and potentially benefit local food security.
- Full Text:
- Date Issued: 2014
Identification of novel marine algal compounds with differential anti-cancer activity: towards a cancer stem-cell specific chemotherapy
- Authors: De la Mare, Jo-Anne
- Date: 2012
- Subjects: Breast -- Cancer , Stem cells -- Research , Chemotherapy , Algae -- Biotechnology
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4143 , http://hdl.handle.net/10962/d1016250
- Description: Breast cancer remains the leading cause of cancer-related death in women worldwide. Furthermore, it has been demonstrated that the treatment-resistant ER-PR-HER2/neu- sub-type is more common among women of African descent, necessitating the search for novel chemotherapies for this form of the disease. The secondary metabolites produced by marine algae represent a rich source of structurally unique compounds with chemotherapeutic potential, particularly in South Africa, whose oceans allegedly host 15 % of the total number of species in the world. Indeed, a recent study reported the isolation of a range of novel compounds from South African red and brown algae of the Plocamium, Portiera and Sargassum genera which displayed cytotoxicity against oesophageal cancer cells in vitro. The molecular mechanisms mediating this toxicity were unknown, as was the effect of these and similar compounds on metastatic ER-PR-HER2/neu- breast cancer cell lines or breast cancer stem cells. The current study aimed to address these questions by screening a library of twenty-two novel marine algal compounds for the ability to inhibit MDA-MB-231 and Hs578T breast cancer cells, while having no adverse effects on non-cancerous MCF12A breast epithelial cells. While twelve of these were toxic in the micromolar range against breast cancer cells, only the polyhalogenated monoterpenes RU004 and RU007, and the tetraprenylated quinone sargaquinoic acid (SQA) were identified as hit compounds based on the criteria that their cytotoxicity was specific to breast cancer and not healthy breast cells in vitro. On the other hand, the halogenated monoterpene RU015 was found to be highly toxic to both breast cancer and non-cancerous breast cell lines, while the halogenated monoterpene stereoisomers RU017 and RU018 were non-toxic to either of these cell lines. The mode of action of RU004, RU007, RU015 and SQA, together with the previously characterized carotenoid fucoxanthin (FXN), was assessed in terms of the type of cell death induced and the effect on cell cycle distribution of these compounds. Flow cytometric analysis of the extent of Hoescht 33342 and propidium iodide staining along with PARP cleavage studies suggested that SQA induced apoptosis in MDA-MB-231 cells. On the other hand, the highly toxic compound RU015 appeared to induce necrosis as evidenced by 50 kDa PARP cleavage product in MDA-MB-231 cells. The flow cytometry profiles of MDA-MB-231 and Hst578T cells treated with the hit compounds RU004 and RU007 were suggestive of the induction of apoptosis by these compounds. Cell cycle analysis by flow cytometry with propidium iodide staining revealed that both SQA and FXN induced G0-G1 arrest together with an increase in the apoptotic sub-G0 population, which agreed with previous reports in the literature. The molecular mechanism of action of SQA and FXN were further investigated by the identification of specific signal transducer molecules involved in mediating their anti-cancer activities. SQA was found to require the activity of numerous caspases, including caspase-3, -6, -8, -9, -10 and -13, for its cytotoxicity and was demonstrated to decrease the level of the antiapoptotic protein Bcl-2. On the other hand, FXN was shown to require caspase-1, -2, -3, -9 and - 10 for its toxicity. This, together with the ability to decrease the levels of Bcl-2, pointed to the involvement of the intrinsic pathway in particular in mediating the activity of FXN. The screening of algal compounds against non-cancerous breast epithelial cells carried out in this study, together with the investigation into their mechanisms of action, represent one of the few reports in which characterization of algal metabolites goes beyond the initial cytotoxicity assays. Finally, in order to assess the potential anti-cancer stem cell activity of the marine algal compounds, a subset of these was screened using a mammosphere assay technique developed in this study. The cancer stem cell (CSC) theory proposes that cancers arise from and are maintained by a specific subpopulation of cells able to undergo asymmetric cell division and termed CSCs. These CSCs are capable of anchorage-independent growth in serum-free culture conditions, such as those in the mammosphere assay. Using this assay, the novel halogenated monoterpene stereoisomers RU017 and RU018 were demonstrated to possess putative anti- CSC activity as evidenced by their ability to completely eliminate mammosphere formation in vitro. Furthermore, since RU017 and RU018 were non-toxic to both breast cancer and healthy breast cells, it appeared that the activity of the compounds was potentially specific to the CSCs. The results require further validation, but represent the first report of selective anti-CSC activity.
- Full Text:
- Date Issued: 2012
- Authors: De la Mare, Jo-Anne
- Date: 2012
- Subjects: Breast -- Cancer , Stem cells -- Research , Chemotherapy , Algae -- Biotechnology
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4143 , http://hdl.handle.net/10962/d1016250
- Description: Breast cancer remains the leading cause of cancer-related death in women worldwide. Furthermore, it has been demonstrated that the treatment-resistant ER-PR-HER2/neu- sub-type is more common among women of African descent, necessitating the search for novel chemotherapies for this form of the disease. The secondary metabolites produced by marine algae represent a rich source of structurally unique compounds with chemotherapeutic potential, particularly in South Africa, whose oceans allegedly host 15 % of the total number of species in the world. Indeed, a recent study reported the isolation of a range of novel compounds from South African red and brown algae of the Plocamium, Portiera and Sargassum genera which displayed cytotoxicity against oesophageal cancer cells in vitro. The molecular mechanisms mediating this toxicity were unknown, as was the effect of these and similar compounds on metastatic ER-PR-HER2/neu- breast cancer cell lines or breast cancer stem cells. The current study aimed to address these questions by screening a library of twenty-two novel marine algal compounds for the ability to inhibit MDA-MB-231 and Hs578T breast cancer cells, while having no adverse effects on non-cancerous MCF12A breast epithelial cells. While twelve of these were toxic in the micromolar range against breast cancer cells, only the polyhalogenated monoterpenes RU004 and RU007, and the tetraprenylated quinone sargaquinoic acid (SQA) were identified as hit compounds based on the criteria that their cytotoxicity was specific to breast cancer and not healthy breast cells in vitro. On the other hand, the halogenated monoterpene RU015 was found to be highly toxic to both breast cancer and non-cancerous breast cell lines, while the halogenated monoterpene stereoisomers RU017 and RU018 were non-toxic to either of these cell lines. The mode of action of RU004, RU007, RU015 and SQA, together with the previously characterized carotenoid fucoxanthin (FXN), was assessed in terms of the type of cell death induced and the effect on cell cycle distribution of these compounds. Flow cytometric analysis of the extent of Hoescht 33342 and propidium iodide staining along with PARP cleavage studies suggested that SQA induced apoptosis in MDA-MB-231 cells. On the other hand, the highly toxic compound RU015 appeared to induce necrosis as evidenced by 50 kDa PARP cleavage product in MDA-MB-231 cells. The flow cytometry profiles of MDA-MB-231 and Hst578T cells treated with the hit compounds RU004 and RU007 were suggestive of the induction of apoptosis by these compounds. Cell cycle analysis by flow cytometry with propidium iodide staining revealed that both SQA and FXN induced G0-G1 arrest together with an increase in the apoptotic sub-G0 population, which agreed with previous reports in the literature. The molecular mechanism of action of SQA and FXN were further investigated by the identification of specific signal transducer molecules involved in mediating their anti-cancer activities. SQA was found to require the activity of numerous caspases, including caspase-3, -6, -8, -9, -10 and -13, for its cytotoxicity and was demonstrated to decrease the level of the antiapoptotic protein Bcl-2. On the other hand, FXN was shown to require caspase-1, -2, -3, -9 and - 10 for its toxicity. This, together with the ability to decrease the levels of Bcl-2, pointed to the involvement of the intrinsic pathway in particular in mediating the activity of FXN. The screening of algal compounds against non-cancerous breast epithelial cells carried out in this study, together with the investigation into their mechanisms of action, represent one of the few reports in which characterization of algal metabolites goes beyond the initial cytotoxicity assays. Finally, in order to assess the potential anti-cancer stem cell activity of the marine algal compounds, a subset of these was screened using a mammosphere assay technique developed in this study. The cancer stem cell (CSC) theory proposes that cancers arise from and are maintained by a specific subpopulation of cells able to undergo asymmetric cell division and termed CSCs. These CSCs are capable of anchorage-independent growth in serum-free culture conditions, such as those in the mammosphere assay. Using this assay, the novel halogenated monoterpene stereoisomers RU017 and RU018 were demonstrated to possess putative anti- CSC activity as evidenced by their ability to completely eliminate mammosphere formation in vitro. Furthermore, since RU017 and RU018 were non-toxic to both breast cancer and healthy breast cells, it appeared that the activity of the compounds was potentially specific to the CSCs. The results require further validation, but represent the first report of selective anti-CSC activity.
- Full Text:
- Date Issued: 2012
The treatment of brewery effluent using an integrated high rate algal ponding system
- Authors: Cilliers, Anneke
- Date: 2012
- Subjects: Water -- Purification -- South Africa , Algae -- Biotechnology , Algae culture , Algae -- Economic aspects
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5331 , http://hdl.handle.net/10962/d1005177 , Water -- Purification -- South Africa , Algae -- Biotechnology , Algae culture , Algae -- Economic aspects
- Description: The application of high rate algal ponds (HRAP) in the treatment of brewery effluent that met the South African Department of Water Affairs and Forestry's (DWAF) general limits for discharge into a natural water resource of 1998 were tested during a lO-month baseline phase, followed by an 11-month optimization phase. The objective of the baseline phase was to monitor the seasonal performance of HRAPs. The hydraulic retention time (HRT) fluctuated between 11.16 d and 12.00 d in HRAPs. The chemical oxygen demand (COD) increased from 130.12 ± 6.94 mg/L (post-AD), to 171.21 ± 7.99 mg/L (post-HRAP) . The presence of algal cells and evaporation contributed towards an increase in post-HRAP COD. The ammonia (NH₄-N) concentration decreased from 46.59 ± 2.47 mg/L (post-AD), to 1.08 ± 0.12 mg/L (post-HRAP). The nitrite (NO₂- N) concentration remained below 1.00 mg/L in post-pilot plant AD, post-PFP and post-HRAP effluent. The phosphate (PO₄-P) concentration decreased from 29.81 ± 1.39 mg/L (post-AD) to 17.30 ± 1.16 mg/L PO₄-P. The objective of the optimization phase was to manipulate the HRT to achieve the maximum treatment rate that met the DWAF general limits for discharge into a natural water resource of 1998. Nitrogen (as NH₄-N, NO₃-N, NO₂-N) removal efficiency was used as an indicator of nutrient removal success. HRT was influenced by season. The optimal HRT for autumn was 4.30 d at a temperature of 20.53ºC in HRAP A2 (heated) and 18.96ºC in HRAP B2 (ambient). The optimal HRT for summer was 2.74 d at 29.90ºC in HRAP A2 (heated) and 26.36ºC in HRAP B2 (ambient). The COD decreased from 152.33 ± 4.85 mg/L (post-AD) to 95 .00 ± 3.75 mg/L (post-HRAP A2), and to 100.82 ± 5.93 mg/L (post-HRAP B2). The incoming NH₄-N concentration decreased from 42.53 ± 1.38 mg/ L (post-AD), to 1.70 ± 0.81 mg/ L (post-HRAP) . The nitrate (NO₃-N) concentration post-HRAP was 12 - 14 mg/L. The main methods for NH₄-N removal were probably NH₄-N volatilization through algal uptake. HRAPs were able to lower nitrogen and phosphorous concentrations to within the DWAF limits under normal operating conditions. It is recommended that HRAP treated brewery wastewater be used for irrigation after salt removal, or alternatively, for groundwater recharge . Regulatory exemptions would be required for higher than permitted COD and EC concentrations to enable these actions.
- Full Text:
- Date Issued: 2012
- Authors: Cilliers, Anneke
- Date: 2012
- Subjects: Water -- Purification -- South Africa , Algae -- Biotechnology , Algae culture , Algae -- Economic aspects
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5331 , http://hdl.handle.net/10962/d1005177 , Water -- Purification -- South Africa , Algae -- Biotechnology , Algae culture , Algae -- Economic aspects
- Description: The application of high rate algal ponds (HRAP) in the treatment of brewery effluent that met the South African Department of Water Affairs and Forestry's (DWAF) general limits for discharge into a natural water resource of 1998 were tested during a lO-month baseline phase, followed by an 11-month optimization phase. The objective of the baseline phase was to monitor the seasonal performance of HRAPs. The hydraulic retention time (HRT) fluctuated between 11.16 d and 12.00 d in HRAPs. The chemical oxygen demand (COD) increased from 130.12 ± 6.94 mg/L (post-AD), to 171.21 ± 7.99 mg/L (post-HRAP) . The presence of algal cells and evaporation contributed towards an increase in post-HRAP COD. The ammonia (NH₄-N) concentration decreased from 46.59 ± 2.47 mg/L (post-AD), to 1.08 ± 0.12 mg/L (post-HRAP). The nitrite (NO₂- N) concentration remained below 1.00 mg/L in post-pilot plant AD, post-PFP and post-HRAP effluent. The phosphate (PO₄-P) concentration decreased from 29.81 ± 1.39 mg/L (post-AD) to 17.30 ± 1.16 mg/L PO₄-P. The objective of the optimization phase was to manipulate the HRT to achieve the maximum treatment rate that met the DWAF general limits for discharge into a natural water resource of 1998. Nitrogen (as NH₄-N, NO₃-N, NO₂-N) removal efficiency was used as an indicator of nutrient removal success. HRT was influenced by season. The optimal HRT for autumn was 4.30 d at a temperature of 20.53ºC in HRAP A2 (heated) and 18.96ºC in HRAP B2 (ambient). The optimal HRT for summer was 2.74 d at 29.90ºC in HRAP A2 (heated) and 26.36ºC in HRAP B2 (ambient). The COD decreased from 152.33 ± 4.85 mg/L (post-AD) to 95 .00 ± 3.75 mg/L (post-HRAP A2), and to 100.82 ± 5.93 mg/L (post-HRAP B2). The incoming NH₄-N concentration decreased from 42.53 ± 1.38 mg/ L (post-AD), to 1.70 ± 0.81 mg/ L (post-HRAP) . The nitrate (NO₃-N) concentration post-HRAP was 12 - 14 mg/L. The main methods for NH₄-N removal were probably NH₄-N volatilization through algal uptake. HRAPs were able to lower nitrogen and phosphorous concentrations to within the DWAF limits under normal operating conditions. It is recommended that HRAP treated brewery wastewater be used for irrigation after salt removal, or alternatively, for groundwater recharge . Regulatory exemptions would be required for higher than permitted COD and EC concentrations to enable these actions.
- Full Text:
- Date Issued: 2012
Analysis of the anti-cancer activity of novel indigenous algal compounds in breast cancer: towards the development of a model for screening anti-cancer stem cell activity
- Authors: Lawson, Jessica Clair
- Date: 2010
- Subjects: Breast -- Cancer , Breast -- Cancer -- Chemotherapy , Breast -- Cancer -- Treatment , Red algae , Brown algae , Algae -- Biotechnology
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3925 , http://hdl.handle.net/10962/d1003984 , Breast -- Cancer , Breast -- Cancer -- Chemotherapy , Breast -- Cancer -- Treatment , Red algae , Brown algae , Algae -- Biotechnology
- Description: Breast cancer, the most common malignancy diagnosed in women, is one of the leading causes of death in women worldwide. In South Africa only 32% of women diagnosed with advanced breast cancer survive more than five years. The search for new chemotherapeutic agents capable of effectively treating breast cancer is therefore essential. Recent evidence supporting the cancer stem cell theory of cancer development for breast cancer challenges the current theories of cancer development and hence treatment. Cancer stem cells are a small subpopulation of tumour cells that possess properties of both cancer cells and stem cells and are believed to be the tumour-initiating population of many cancers. Cancer stem cells are inherently resistant to many chemotherapeutic agents and in this way have been associated with repopulation of tumours after chemotherapy. This phenomenon is proposed as a possible mechanism for cancer relapse after treatment. Cancer stem cells have also been implicated in metastasis, the major cause of mortality in cancer patients. Therefore, any treatment that is capable of targeting and removing breast cancer stem cells may have the theoretical potential to effectively treat breast cancer. However, there are currently no such treatments available for clinical use. We were provided access to a library of novel indigenous small molecules isolated from red and brown algae found off the Eastern Cape of South Africa. The aim of this project was to analyse the anti-cancer and anti-cancer stem cell properties of the compounds in this library and to identify „hit‟ compounds which could form the basis for future development into new anti-cancer drugs. Ten novel compounds of algal origin were tested for cytotoxicity, by determining their ability to inhibit the growth of MCF12A breast epithelial cells and MCF7 breast cancer cells using the colorimetric MTT [(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] cell proliferation assay. All but one of the compounds tested exhibited cytotoxicity towards the MCF7 cancer cell line, with IC50 values (the concentration of the compound that leads to a 50% inhibition in cell growth) of between 3 μM and 90 μM. The chemotherapeutic drug paclitaxel was used as a positive control. Four of the compounds (RUMB-001, RUMB-002, RUMB-007 and RUMB-010/saragaquinoic acid) were significantly more toxic to the MCF7 cancer cell line, than the „normal‟ MCF12A breast cells and were selected as priority compounds for further analyses. In addition, two other compounds were selected as priority compounds, one highly cytotoxic towards both MCF12A and MCF7 cell lines (RUMB-015) and one which was non toxic to either cell line (RUMB-017/018). Preliminary studies into the mechanism of cytotoxicity using Western blot analysis for poly (ADP-ribose) polymerase (PARP) cleavage and Hoechst 33342 immunostaining in MCF-7 cells were largely unsuccessful. The Hoechst 33342 immunostaining assay did provide tentative evidence that selected priority compounds were capable of inducing apoptosis, although these assays will need to be repeated using a less subjective assay to confirm the results. The priority compounds were subsequently investigated for their cytotoxic effect on the cancer stem cell-enriched side population in MCF7 cells. The ability of the priority compounds to selectively target the cancer stem cell containing side population was assessed using two complementary flow cytometry-based techniques – namely the Hoechst 33342-exclusion assay, and fluorescent immunostaining for the expression of the putative cancer stem cell marker, ABCG2+. The ABCG2+ staining assay was a novel technique developed during the course of this study. It remains to be fully validated, but it may provide a new and reliable way to identify and analyse cancer stem cell containing side population cells. The MCF7 cells were treated with the compounds and the proportion of putative cancer stem cells compared with the size of the population in untreated cells was assessed. Three compounds (RUMB-010, RUMB-015 and RUMB-017/018) capable of reducing the proportion of side population cells within the MCF7 cell line were identified. Taking these data together, we identified two potential „hit‟ compounds which should be prioritised for future research. These are compounds RUMB-010/sargaquinoic acid and RUMB-017/018. RUMB-010 is of interest as it was shown to target the putative cancer stem cell population, in addition to the bulk MCF7 tumour line, but was relatively less toxic to the „normal‟ MCF12A cell line. RUMB-017/018 is of interest due to the ability to selectively target the cancer stem cell enriched side population, while having little effect on the normal (MCF12A) or bulk tumour (MCF7) cell lines tested. These compounds will be important as „hit‟ compounds for drug development and as tool compounds to study cancer and cancer stem cell biology.
- Full Text:
- Date Issued: 2010
- Authors: Lawson, Jessica Clair
- Date: 2010
- Subjects: Breast -- Cancer , Breast -- Cancer -- Chemotherapy , Breast -- Cancer -- Treatment , Red algae , Brown algae , Algae -- Biotechnology
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3925 , http://hdl.handle.net/10962/d1003984 , Breast -- Cancer , Breast -- Cancer -- Chemotherapy , Breast -- Cancer -- Treatment , Red algae , Brown algae , Algae -- Biotechnology
- Description: Breast cancer, the most common malignancy diagnosed in women, is one of the leading causes of death in women worldwide. In South Africa only 32% of women diagnosed with advanced breast cancer survive more than five years. The search for new chemotherapeutic agents capable of effectively treating breast cancer is therefore essential. Recent evidence supporting the cancer stem cell theory of cancer development for breast cancer challenges the current theories of cancer development and hence treatment. Cancer stem cells are a small subpopulation of tumour cells that possess properties of both cancer cells and stem cells and are believed to be the tumour-initiating population of many cancers. Cancer stem cells are inherently resistant to many chemotherapeutic agents and in this way have been associated with repopulation of tumours after chemotherapy. This phenomenon is proposed as a possible mechanism for cancer relapse after treatment. Cancer stem cells have also been implicated in metastasis, the major cause of mortality in cancer patients. Therefore, any treatment that is capable of targeting and removing breast cancer stem cells may have the theoretical potential to effectively treat breast cancer. However, there are currently no such treatments available for clinical use. We were provided access to a library of novel indigenous small molecules isolated from red and brown algae found off the Eastern Cape of South Africa. The aim of this project was to analyse the anti-cancer and anti-cancer stem cell properties of the compounds in this library and to identify „hit‟ compounds which could form the basis for future development into new anti-cancer drugs. Ten novel compounds of algal origin were tested for cytotoxicity, by determining their ability to inhibit the growth of MCF12A breast epithelial cells and MCF7 breast cancer cells using the colorimetric MTT [(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] cell proliferation assay. All but one of the compounds tested exhibited cytotoxicity towards the MCF7 cancer cell line, with IC50 values (the concentration of the compound that leads to a 50% inhibition in cell growth) of between 3 μM and 90 μM. The chemotherapeutic drug paclitaxel was used as a positive control. Four of the compounds (RUMB-001, RUMB-002, RUMB-007 and RUMB-010/saragaquinoic acid) were significantly more toxic to the MCF7 cancer cell line, than the „normal‟ MCF12A breast cells and were selected as priority compounds for further analyses. In addition, two other compounds were selected as priority compounds, one highly cytotoxic towards both MCF12A and MCF7 cell lines (RUMB-015) and one which was non toxic to either cell line (RUMB-017/018). Preliminary studies into the mechanism of cytotoxicity using Western blot analysis for poly (ADP-ribose) polymerase (PARP) cleavage and Hoechst 33342 immunostaining in MCF-7 cells were largely unsuccessful. The Hoechst 33342 immunostaining assay did provide tentative evidence that selected priority compounds were capable of inducing apoptosis, although these assays will need to be repeated using a less subjective assay to confirm the results. The priority compounds were subsequently investigated for their cytotoxic effect on the cancer stem cell-enriched side population in MCF7 cells. The ability of the priority compounds to selectively target the cancer stem cell containing side population was assessed using two complementary flow cytometry-based techniques – namely the Hoechst 33342-exclusion assay, and fluorescent immunostaining for the expression of the putative cancer stem cell marker, ABCG2+. The ABCG2+ staining assay was a novel technique developed during the course of this study. It remains to be fully validated, but it may provide a new and reliable way to identify and analyse cancer stem cell containing side population cells. The MCF7 cells were treated with the compounds and the proportion of putative cancer stem cells compared with the size of the population in untreated cells was assessed. Three compounds (RUMB-010, RUMB-015 and RUMB-017/018) capable of reducing the proportion of side population cells within the MCF7 cell line were identified. Taking these data together, we identified two potential „hit‟ compounds which should be prioritised for future research. These are compounds RUMB-010/sargaquinoic acid and RUMB-017/018. RUMB-010 is of interest as it was shown to target the putative cancer stem cell population, in addition to the bulk MCF7 tumour line, but was relatively less toxic to the „normal‟ MCF12A cell line. RUMB-017/018 is of interest due to the ability to selectively target the cancer stem cell enriched side population, while having little effect on the normal (MCF12A) or bulk tumour (MCF7) cell lines tested. These compounds will be important as „hit‟ compounds for drug development and as tool compounds to study cancer and cancer stem cell biology.
- Full Text:
- Date Issued: 2010
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 removal of toxic heavy metals from aqueous solutions by algal extracellular polysaccharides
- Authors: Selepe, Mamaropeng Marcus
- Date: 1999
- Subjects: Heavy metals -- Absorption and adsorption , Copper , Lead , Algae -- Biotechnology , Polysaccharides -- Biotechnology
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3994 , http://hdl.handle.net/10962/d1004054 , Heavy metals -- Absorption and adsorption , Copper , Lead , Algae -- Biotechnology , Polysaccharides -- Biotechnology
- Description: This study investigated the possible use of algal extracellular polysaccharide as a biosorbent for removal of heavy metals (copper and lead) from aqueous solutions as a means of bioremediation for metal containing effluents. This biopolymer has good biosorbent properties and a potential to provide a cost effective, selective and efficient purification system. A variety of environmental conditions induce the production of extracellular polysaccharides in algae. The production of exopolysaccharides by Dunaliella cultures was induced by nitrogen deficient conditions. A high ratio of carbon to nitrogen source considerably enhanced the polysaccharide release. Purified extracellular polysaccharide samples exhibited a monosaccharide composition consisting of the following sugars: xylose, arabinose, 2-0-methyl mannose, mannose, glucose and galactose. The relative abundance (%) of these sugars were calculated relative to xylose. The major sugar constituent was 2-0-methyl mannose, which was present at approximately 160% relative to xylose. The percentage relative abundance of other sugars was as follows: 18.8; 86.8; 85.3 and 22.3% for arabinose; mannose; glucose and galactose respectively. The identity of the various constituents were confirmed by mass spectrometry. The ability of Dunaliella exopolysaccharides to accumulate metals was investigated. The following parameters were studied because they affect metal uptake: solution pH, biomass concentration, temperature, time and metal concentration. The uptake of both copper and lead were pH dependent. However, metal uptake was not significantly affected by temperature. Kinetic studies showed that Dunaliella extracellular polysaccharides exhibit good bioremediation properties. Metal uptake was rapid. In addition, the exopolysaccharide has good metal binding capacity with an uptake capacity for lead of 80 mg/g from a solution containing initial lead concentration of approximately 40 mg/l. Competition studies revealed that the presence of a second metal in solution inhibits uptake of the other metal compared to uptake in single metal solution of that particular metal. The presence of lead inhibited the uptake of copper from approximately 65% in single metal solution to 10% in binary metal solution. The presence of copper also inhibited lead uptake, though not to the same extent. Higher concentrations of lead could not completely prevent removal of copper from solution and visa versa. The same was true for lead which could not be displaced by a four-fold concentration of copper. Instead, a certain percentage of copper was always removed showing that lead did not compete with copper for these binding sites. In conclusion it appears that, copper and lead bind to different sites on Dunaliella exopolysaccharides and that they exhibit selective or preferential removal of lead.
- Full Text:
- Date Issued: 1999
- Authors: Selepe, Mamaropeng Marcus
- Date: 1999
- Subjects: Heavy metals -- Absorption and adsorption , Copper , Lead , Algae -- Biotechnology , Polysaccharides -- Biotechnology
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3994 , http://hdl.handle.net/10962/d1004054 , Heavy metals -- Absorption and adsorption , Copper , Lead , Algae -- Biotechnology , Polysaccharides -- Biotechnology
- Description: This study investigated the possible use of algal extracellular polysaccharide as a biosorbent for removal of heavy metals (copper and lead) from aqueous solutions as a means of bioremediation for metal containing effluents. This biopolymer has good biosorbent properties and a potential to provide a cost effective, selective and efficient purification system. A variety of environmental conditions induce the production of extracellular polysaccharides in algae. The production of exopolysaccharides by Dunaliella cultures was induced by nitrogen deficient conditions. A high ratio of carbon to nitrogen source considerably enhanced the polysaccharide release. Purified extracellular polysaccharide samples exhibited a monosaccharide composition consisting of the following sugars: xylose, arabinose, 2-0-methyl mannose, mannose, glucose and galactose. The relative abundance (%) of these sugars were calculated relative to xylose. The major sugar constituent was 2-0-methyl mannose, which was present at approximately 160% relative to xylose. The percentage relative abundance of other sugars was as follows: 18.8; 86.8; 85.3 and 22.3% for arabinose; mannose; glucose and galactose respectively. The identity of the various constituents were confirmed by mass spectrometry. The ability of Dunaliella exopolysaccharides to accumulate metals was investigated. The following parameters were studied because they affect metal uptake: solution pH, biomass concentration, temperature, time and metal concentration. The uptake of both copper and lead were pH dependent. However, metal uptake was not significantly affected by temperature. Kinetic studies showed that Dunaliella extracellular polysaccharides exhibit good bioremediation properties. Metal uptake was rapid. In addition, the exopolysaccharide has good metal binding capacity with an uptake capacity for lead of 80 mg/g from a solution containing initial lead concentration of approximately 40 mg/l. Competition studies revealed that the presence of a second metal in solution inhibits uptake of the other metal compared to uptake in single metal solution of that particular metal. The presence of lead inhibited the uptake of copper from approximately 65% in single metal solution to 10% in binary metal solution. The presence of copper also inhibited lead uptake, though not to the same extent. Higher concentrations of lead could not completely prevent removal of copper from solution and visa versa. The same was true for lead which could not be displaced by a four-fold concentration of copper. Instead, a certain percentage of copper was always removed showing that lead did not compete with copper for these binding sites. In conclusion it appears that, copper and lead bind to different sites on Dunaliella exopolysaccharides and that they exhibit selective or preferential removal of lead.
- Full Text:
- Date Issued: 1999
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
Algal biotechnology and the beneficiation of saline effluent wastes
- Authors: Rose, P D (Peter Dale)
- Date: 1992
- Subjects: Algae -- Biotechnology , Algae culture , Tanneries -- Waste disposal
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4135 , http://hdl.handle.net/10962/d1015967
- Description: Saline deterioration in the South African public water system has been documented and disposal of brine wastes has been identified as part of the problem. The broad aim of this research programme was to undertake an initial technical study to evaluate the feasibility of integrating algal biotechnology into a disposal function for these wastes. A demonstration of utility in the form of products and waste treatment could produce a beneficiation of saline effluents and provide incentives necessary to deal with the disposal issue. The study attempted to demonstrate a synthesis between the two main thrusts in algal biotechnology that have produced large-scale practical applications - stable, predictable algal production in saline media and the cost effective High Rate Oxidation Ponding (HROP) process for incorporating algal production into a waste treatment function. Tannery organic saline effluents and the biotechnology of Dunaliella salina culture producing β- carotene were chosen as paradigms for the study. 1. The alga was shown to grow in certain tannery effluents producing enhanced biomass yields compared to defined inorganic medium cultivation. The potential for amino acid or protein supplementation of defmed culture media was noted. 2. A reduction in organic load simultaneous with the growth of D.salina was recorded in laboratory-scale simulations of the HROP process. Rates similar to the fresh water HROP equivalent were demonstrated. 3. These results suggested the uptake and storage of organic nitrogen by D.salina. The consequent inhibition of β-carotene accumulation by the organism presented a potentially insurmountable obstacle to the feasibility of β-carotene production in this medium. Uptake and release of organic compounds, previously demonstrated in phytoplankton and other micro-algae, was confirmed in this study for D.salina. The evidence acquired indicated the internalization of both glycine and bovine serum albumin. An ultrastructural study demonstrated mechanisms by which this process might occur. 4. The release of substantial quantities of glycerol was shown. A mechanism whereby D. salina may use this to regulate ammonia availability via control of its associated bacterial population was observed. Glycerol release was identified as presenting an application in treating refractory organic wastes, such as secondary sewage sludges, by elevating C:N ratios. This could demonstrate a significant utility for brine waste impoundments. 5. A multistage production process was proposed to deal with the problem of β-carotene inhibition by separation of the growth and metabolite accumulation functions into separate unit operations. It was shown in this study that the stress of nitrogen deficiency combined with high salinity provides for effectiveβ-carotene accumulation under the conditions of low illumination that pertain in dense cultures. Subjected to these conditions effluent-grown cells show delayed but unimpaired {j-carotene accumulation. 6. A role for the plant hormone abscisic acid in mediating the stress response was demonstrated in D.salina. Fluorescence induction studies suggested the presence of a signalling process forming part of a sensitivity control mechanism. Stress induction of β-carotene accumulation could occur through four clearly defined stages. Potential was identified for using this response as a physiological probe for monitoring and regulating the stress induction process. 7. The multistage processing concept requires effective algal cell separation technology. The use of cross-flow ultrafiltration and diafiltration with a polyethersulfone tubular membrane system was demonstrated as an effective process for the recovery and washing of D. salina. Cell concentrates were produced in a viable form. 8. Process designs incorporating the findings of the research programme are presented demonstrating how effluent and organic waste treatment functions may be combined with the production of D.salina and its products. Application of the multi-stage processing concept to β-carotene production in a defined medium process was identified as offering a potential four-fold yield enhancement. This could have a significant impact on a high cost, marginal algal biotechnology process. Aspects of novelty have been claimed in provisional patents applications. A provisional demonstration of the feasibility of D.salina production in tannery effluent indicates that algal biotechnology may provide a utility for, and hence the beneficiation of saline effluent wastes.
- Full Text:
- Date Issued: 1992
- Authors: Rose, P D (Peter Dale)
- Date: 1992
- Subjects: Algae -- Biotechnology , Algae culture , Tanneries -- Waste disposal
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4135 , http://hdl.handle.net/10962/d1015967
- Description: Saline deterioration in the South African public water system has been documented and disposal of brine wastes has been identified as part of the problem. The broad aim of this research programme was to undertake an initial technical study to evaluate the feasibility of integrating algal biotechnology into a disposal function for these wastes. A demonstration of utility in the form of products and waste treatment could produce a beneficiation of saline effluents and provide incentives necessary to deal with the disposal issue. The study attempted to demonstrate a synthesis between the two main thrusts in algal biotechnology that have produced large-scale practical applications - stable, predictable algal production in saline media and the cost effective High Rate Oxidation Ponding (HROP) process for incorporating algal production into a waste treatment function. Tannery organic saline effluents and the biotechnology of Dunaliella salina culture producing β- carotene were chosen as paradigms for the study. 1. The alga was shown to grow in certain tannery effluents producing enhanced biomass yields compared to defined inorganic medium cultivation. The potential for amino acid or protein supplementation of defmed culture media was noted. 2. A reduction in organic load simultaneous with the growth of D.salina was recorded in laboratory-scale simulations of the HROP process. Rates similar to the fresh water HROP equivalent were demonstrated. 3. These results suggested the uptake and storage of organic nitrogen by D.salina. The consequent inhibition of β-carotene accumulation by the organism presented a potentially insurmountable obstacle to the feasibility of β-carotene production in this medium. Uptake and release of organic compounds, previously demonstrated in phytoplankton and other micro-algae, was confirmed in this study for D.salina. The evidence acquired indicated the internalization of both glycine and bovine serum albumin. An ultrastructural study demonstrated mechanisms by which this process might occur. 4. The release of substantial quantities of glycerol was shown. A mechanism whereby D. salina may use this to regulate ammonia availability via control of its associated bacterial population was observed. Glycerol release was identified as presenting an application in treating refractory organic wastes, such as secondary sewage sludges, by elevating C:N ratios. This could demonstrate a significant utility for brine waste impoundments. 5. A multistage production process was proposed to deal with the problem of β-carotene inhibition by separation of the growth and metabolite accumulation functions into separate unit operations. It was shown in this study that the stress of nitrogen deficiency combined with high salinity provides for effectiveβ-carotene accumulation under the conditions of low illumination that pertain in dense cultures. Subjected to these conditions effluent-grown cells show delayed but unimpaired {j-carotene accumulation. 6. A role for the plant hormone abscisic acid in mediating the stress response was demonstrated in D.salina. Fluorescence induction studies suggested the presence of a signalling process forming part of a sensitivity control mechanism. Stress induction of β-carotene accumulation could occur through four clearly defined stages. Potential was identified for using this response as a physiological probe for monitoring and regulating the stress induction process. 7. The multistage processing concept requires effective algal cell separation technology. The use of cross-flow ultrafiltration and diafiltration with a polyethersulfone tubular membrane system was demonstrated as an effective process for the recovery and washing of D. salina. Cell concentrates were produced in a viable form. 8. Process designs incorporating the findings of the research programme are presented demonstrating how effluent and organic waste treatment functions may be combined with the production of D.salina and its products. Application of the multi-stage processing concept to β-carotene production in a defined medium process was identified as offering a potential four-fold yield enhancement. This could have a significant impact on a high cost, marginal algal biotechnology process. Aspects of novelty have been claimed in provisional patents applications. A provisional demonstration of the feasibility of D.salina production in tannery effluent indicates that algal biotechnology may provide a utility for, and hence the beneficiation of saline effluent wastes.
- Full Text:
- Date Issued: 1992
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