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
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- Date Issued: 2021-10-29
Characterization of bioflocculants produced by consortia of three marine bacteria belonging to the genera bacillus and cobetia previously isolated from the bottom sediment of Algoa Bay in the Eastern Cape Province of South Africa
- Authors: Ugbenyen, Anthony Moses https://orcid.org/0000-0002-1381-3428
- Date: 2013
- Subjects: Water -- Purification -- Flocculation , Water quality management , Flocculation
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10353/24454 , vital:62804
- Description: The bioflocculant-producing potentials of three marine bacteria isolated from the sediment samples of Algoa Bay in the Eastern Cape Province of South Africa were assessed. Analysis of the partial nucleotide sequence of the 16S rDNA of the bacteria revealed 99 percent, 99 percent, and 98 percent similarity to Cobetia sp. L222, Bacillus sp. A-5A, and Bacillus sp. HXG-C1 respectively and the sequence was deposited in GenBank as Cobetia sp. OAUIFE, Bacillus sp. MAYA and Bacillus sp. Gilbert (accession number JF799092, JF799093, and HQ537128 respectively). Cultivation condition studies for Cobetia sp. OAUIFE revealed that bioflocculant production was optimal with an inoculum size of 2 percent (v/v), initial pH of 6.0, Mn2+ as the metal ion, and glucose as the carbon source. Metal ions, including Na+, K+, Li+, Ca2+and Mg2+ stimulated bioflocculant production resulting in flocculating activity of above 90 percent. This crude bioflocculant is thermally stable, with about 78 percent of its flocculating activity remaining after heating at 100 oC for 25 min. Analysis of the purified bioflocculant revealed it to be an acidic extracellular polysaccharide. FTIR analysis revealed the presence of methoxyl, hydroxyl, and carboxyl - groups in the compound bioflocculant and SEM micrograph of the bioflocculant revealed a crystal-linear structure. On the other hand, bioflocculant production by Bacillus sp. MAYA was optimal when glucose (95.6 percent flocculating activity) and ammonium nitrate (83.3 percent flocculating activity) were used as carbon and nitrogen sources respectively; inoculum size was 2 percent (v/v); initial pH 6; and Ca2+ as coagulant aid. Chemical analysis of the purified bioflocculant shows that it is composed of uronic acid, neutral sugar and protein. FTIR analysis also revealed the presence of methoxyl, hydroxyl, carboxyl and amino- groups in this bioflocculant. The bioflocculant is thermostable with about 65.6 percent residual flocculating activity retained after heating the bioflocculant at 100 oC for 25 min. However bioflocculant production by Bacillus sp. Gilbert was optimal when sodium carbonate (95.2 percent flocculating activity) and potassium nitrate (76.6 percent flocculating activity) were used as carbon and nitrogen sources respectively; inoculum size was 3 percent (v/v); initial pH 9; and Al3+ as cation. The crude bioflocculant retained 44.2 percent residual flocculating activity after heating at 100 oC for 15 min. FTIR analysis reveals the presence of hydroxyl, carboxyl and methylene - groups in the compound bioflocculant. SEM micrograph of the bioflocculant revealed an amorphous compound. The consortia of these bacteria strains also produced bioflocculants with high flocculating activities which were highly efficient in removing turbidity and chemical oxygen demand (COD) from brewery wastewater, diary wastewater and river water. The bioflocculants from the consortia seemed better than traditional flocculants such as alum . The characteristics of the bioflocculant produced by the consortium of Cobetia sp. OAUIFE and Bacillus sp. MAYA showed that this extracellular bioflocculant, composed of 66percent uronic acid and 31percent protein and an optimum flocculation (90 percent) of kaolin suspension, when the dosage concentration was 0.8 mg/ml, under weak alkaline pH of 8, and Ca2+ as a coagulant aid. The bioflocculant is thermally stable, with a high residual flocculating activity of 86.7 percent, 89.3 percent and 87.0 percent after heating at 50 oC, 80 oC and 100 oC for 25 min respectively. The FTIR analysis of the bioflocculant indicated the presence of hydroxyl, amino, carbonyl and carboxyl functional groups. Scanning electron microscopy (SEM) image revealed a crystal-linear spongy-like bioflocculant structure and EDX analysis of the purified bioflocculant showed that the elemental composition in mass proportion of C,N,O,S and P was 6.67:6.23:37.55:0.38:4.42 (percent w/w). However, the characteristics of the bioflocculant produced by the consortium of Cobetia sp OAUIFE and Bacillus sp. Gilbert showed an optimum flocculation (90 percent) of kaolin suspension when the dosage concentration was 0.2 mg/ml, under neutral pH of 7, and Ca2+ as a coagulant aid. The FTIR analysis of the bioflocculant indicated the presence of hydroxyl and carbonyl functional groups. Scanning electron microscopy (SEM) image revealed an amorphous morphology. On the other hand the bioflocculant produced by the consortium of Bacillus sp. MAYA and Bacillus sp. Gilbert showed similar characteristic with the bioflocculant from the consortium of Cobetia sp. OAUIFE and Bacillus sp. Gilbert except for Al3+ being the preferred coagulant aid. The characteristics of the bioflocculant produced by the consortium of Cobetia sp. OAUIFE, Bacillus sp. MAYA and Bacillus sp. Gilbert showed an optimum flocculation (87 percent) of kaolin suspension when the dosage concentration was 1.0 mg/ml. Under strong alkaline pH of 12, flocculating activity reached (95 percent) when Al3+ was the coagulant aid. The FTIR analysis of the bioflocculant indicated the presence of hydroxyl, amino, carbonyl and carboxyl and phosphoryl functional groups. Scanning electron microscopy (SEM) image revealed a flaky amorphous morphological structure. Due to the excellent COD and turbidity removal efficiencies of the bioflocculants produced by the consortia, these make those attractive candidates for use in water and wastewater treatment. , Thesis (PhD) -- Faculty of Science and Agriculture, 2013
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- Date Issued: 2013
Production and biochemical characterization of new bioflocculants from bacteria isolated from freshwater and marine environments of the Eastern Cape in South Africa
- Authors: Mabinya, Leonard Vuyani https://orcid.org/0000-0002-0682-7282
- Date: 2013-01
- Subjects: Flocculation , Bacteria
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10353/24228 , vital:62445
- Description: The production and characterization of bioflocculants produced by three bacteria belonging to Arthrobacter, Halomonas and Micrococcus genera and isolated from freshwater and marine environments were evaluated both as axenic cultures and as consortia. The influences of cultutre conditions such as carbon, nitrogen and metal ions sources, as well as initial pH on bioflocculant production by individual isolates were investigated. Both Arthrobacter sp. Raats and Halomonas sp. Okoh utilized urea as a nitrogen source of choice for optimal production of the bioflocculants with Micrococcus sp. Leo having a preference for peptone. All three strains differed in as far as the carbon source of choice was concerned with lactose, glucose and sucrose the preferred carbon sources respectively. Also, all three bacterial strains produced an extracellular bioflocculant aerobically but an intial pH 7.0 of the culture media was suitable for both Arthrobacter sp. Raats and Halomonas sp. Okoh with a slightly alkaline pH of 9.0 preferred by Micrococcus sp. Leo. The presence of Mg2+ cations stimulated bioflocculant production by both Arthrobacter sp. Raats and Micrococcus sp. Leo while Ca2+ resulted in more efficient bioflocculant production by Halomonas sp. Okoh. Chemical analyses revealed the bioflocculants produced by both Halomonas sp. Okoh and Micrococcus sp. Leo to be predominantly polysaccharides whereas Arthrobacter sp. Raats produced principally a glycoprotein composed of about 56percent protein and 25percent total carbohydrate. Response surface methodology (RSM) was used to optimize production medium for bioflocculant production by a consortium of Halomonas sp. Okoh and Micrococcus sp. Leo. Plackett-Burman experimental design showed that fructose, ammonium sulphate and MgCl2 were significant in the high yield of the bioflocculant. Furthermore, central composite design showed that optimal concentration of these critical nutritional sources were 16.14 g/L, 1.55 g/L and 1.88 g/L for fructose, ammonium sulphate and MgCl2 respectively. Quantification of the bioflocculant showed a yield of 6.43 g/L which was in close accord with the predicted value of 6.51 g/L. FTIR spectrometry of the bioflocculant indicated the presence of carboxyl, hydroxyl and amino groups, typical for heteropolysaccharide, while SEM imaging revealed a lattice-like structure. The efficiency of the nutrient optimization suggests suitability for industrial applicability. , Thesis (PhD) -- Faculty of Science and Agriculture, 2013
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- Date Issued: 2013-01
Studies on bioflocculant production by a consortium of two bacterial species belonging to the Methylobacterium and Actinobacterium genera
- Authors: Ntsaluba, Luvuyo
- Date: 2012
- Subjects: Flocculation , Actinobacteria , Methylobacterium , Water -- Purification
- Language: English
- Type: Thesis , Masters , MSc (Microbiology)
- Identifier: vital:11266 , http://hdl.handle.net/10353/482 , Flocculation , Actinobacteria , Methylobacterium , Water -- Purification
- Description: Bioflocculants produced by two identified bacteria: Actinobacterium sp. Mayor and Methylobacterium sp. Obi were investigated with regard to their physicochemical and flocculating characteristics. The two strains were later combined to form a consortium for further studies. The optimum culture conditions for the bioflocculant production were similar for all strains except in the case of Actinobacterium sp. Mayor and the consortium, where glucose was replaced by sodium carbonate as a carbon source. Multi-nitrogen source was the best nitrogen source compare to individual sources for both strains. The divalent cation, Ca2+ proved to be a better flocculating activity stimulus for all produced bioflocculants in this study. The optimum flocculating activities obtained for both individual strains and the consortium were all at alkaline pH. The yield of purified bioflocculant produced by the consortium was 8.203 g/l, while 4.190 g/l and 4.610 g/l were recovered for single strains of Actinobacterium sp. Mayor and Methylobacterium sp. Obi respectively. Further characterization of pure bioflocculants revealed that a bioflocculant dosage of 0.3 mg/ml resulted in the highest flocculating activity for both individual strains while 1.0 mg/ml of the bioflocculant produced by the consortium was required to enhance maximum flocculating efficiency. These bioflocculants proved to be all thermo stable at a temperature range of 20 to 900°C with a heating rate of 10oC/min under a constant flow of nitrogen gas. The presence of functional groups normally required for bioflocculation such as hydroxyl, carboxyl and amino was also detected. The findings of this study suggest that the producedbioflocculants can be utilized as excellent substitutes for harmful synthetic flocculants in both water and wastewater treatments as well as in other industrial applications.
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- Date Issued: 2012