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Evaluation of bacteria laccase hybrid biosensor and application in the detection of phenolic contaminants in water

  • Authors: Edoamodu, Chiedu Epiphany https://orcid.org/0000-0002-9254-3955
  • Date: 2021-09
  • Subjects: Laccase , Water -- Purification
  • Language: English
  • Type: Doctoral theses , text
  • Identifier: http://hdl.handle.net/10353/22820 , vital:52802
  • Description: The continuous outpour of aromatic pollutants in diverse groups, emanating from the industrial and domestic system due to population density, industrialisation and technological advancement is alarming. The increasing strength in wastewater discharge constitutes the main cause of the natural water pollution load, causing scarcity of potable water for consumption with an increasing health challenge. The physiochemical treatment approach has faced a series of limitations with little or no success. Hindrance to wastewater management can cause a point source contamination problem that might increase treatment cost and release a broad range of chemical contaminants in the environment. Hence, green, eco-friendly and cost-effective tools are imperative. The application of laccase has received much attention in bioremediation and bioprocessing matters owing to the oxidising capacity of a wide range of substrates. The process requires available molecular oxygen for its activation, releasing water as a by-product thus, establishing this research. This study was devised to examine the bioprocessing potentials of hybrid and amalgamated laccases extracted from the diverse environmental milieu of the Eastern Cape Province, South Africa. Bacteria producing laccase were isolated from marine sediment, cow dung, and wastewater samples via selective enrichment with some aromatic compounds. The axenic cultures were screened for laccase activity on various phenolic and non-phenolic substrates. The isolates were identified via molecular techniques and they belonged to the gammaproteobacteria and Bacilli classes under the following genera, Enterobacter and Bacillus. They were deposited in the NCBI database as Enterobacter asburiae ES1, Enterobacter sp. Kamsi, Enterobacter sp. AI1, and Bacillus sp. NU2 with the assigned accession number MN686602, MN686603, MN686605, MN686607, respectively. Optimisation of the laccase production via one factor at a time technique (OFAT) from the four bacteria species showed more enzyme yield in all lignocellulosic agro-waste media. However, wheat bran and mandarin peel maximally enhanced laccase production. In addition, xylose, galactose, fructose, and sorbitol were the best carbon sources utilised while (NH₄)₂SO8, KNO3 and NaNO3 were noted as the best nitrogen sources employed. Laccase yields were increased at pH 4 and 5, at temperatures 45 and 55 o C, and at 50 and 100 rpm, and precisely, at day eight of the incubation period. Further purification of the crude laccase yielded a purification fold of 4.18, 4.39, 2.78, 8.11, and the SDS-PAGE analysis showed a molecular size of 90, 55, 75 and 50 kDa for ES1, Kamsi, AI1, and NU2 laccases, respectively. The characterised purified laccase demonstrated polyextremotolerant potentials. The laccases were active through a wide temperature regime (30-90 o C) with maximum activity at 50 o C (ES1/AI1 and Kamsi/NU2) 60 o C (AI1), 70 o C (ES1, Kamsi, NU2); and were stable at 60 o C (ES1, AI1, NU2), 70 o C (ES1/AI1), 80 o C (Kamsi and Kamsi/NU2). Also, the laccases remained active through pH 3 - 8 and optimal at pH 4 (AI1, NU2), pH 5 (Kamsi, ES1/AI1), pH 7 (ES1), pH 8 (Kamsi/NU2), and the individual stability was measured at pH 4 (Kamsi, NU2), pH 5 (AI1), pH 6 (ES1), pH 7 (Kamsi/NU2), pH 10 (ES1/AI1). The purified laccases were either enhanced or left unchanged by a variable concentration of metallic salts, inhibitors, chelating agents and organic solvents. Clearly, the activities of the laccase were enhanced when pre-incubated with 1, 3, and 6 mm of CuCl2, FeCl3, MgCl2, ZnCl2 and AgCl, and 1, 2, 3 mm of Triton x-100, PMSF, EDTA, Tween 20, and NaCl. Additionally, 20, 30, and 10 percent v/v of acetone and DMSO were prominent organic solvents that also stimulated both the hybrid and amalgamated laccase activity. The gene of the purified laccases targeted showed a clear band size of 690 bp for the Enterobacter species laccases and 775 bp for the laccase from Bacillus sp. The protein sequence was deposited in NCBI database with the assigned accession numbers, MW251989, MW25990, MW251992, and MW251994 for ES1, Kamsi, AI1, and NU2 laccases, respectively. The optimised pH and temperature parameter examined on the decolourising potential of the bacteria laccases showed an effective dye removal on the five synthetic dyes (Congo Red (CR), Methyl Orange (MO), Malachite Green (MG), Reactive Blue 4 (RB4), Ramazol Brilliant Blue R (RBBR)) applied. The purified laccases were successfully immobilised in Na-alginate with cca. 88.49, 70.91, 76.04, 76.13, 90.07, and 91.99 laccase yield for the hybrid (ES1, ES1, Kamsi, AI1, NU2) and amalgamated (ES1/AI1 and Kamsi/NU2) laccases. The immobilised laccases were able to retain an average activity of 32 – 52 percent after eight dye decolourising cycles, exhibiting strong catalytic activity than the free laccases. Nonetheless, no significant difference was examined between the hybrid and amalgamated laccase activity. Also, the immobilised laccases were shown to be more efficient in biotechnological application than the free laccases. The result suggests that immobilising an enzyme in a carrier matrix served effectively as the remediation approach than the hybrid and the amalgamation of the free enzymes. Also, the application of lignocellulosic waste served as a cheaper substrate for commercial production of laccase and could help s in promoting es the biotechnology application and the bioeconomy. , Thesis (PhD) -- Faculty of Science and Agriculture, 2021
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  • Date Issued: 2021-09

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).
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  • Date Issued: 2020

Synthesis of indium phthalocyanines for photodynamic antimicrobial chemotherapy and photo-oxidation of pollutants

  • Authors: Sindelo, Azole
  • Date: 2019
  • Subjects: Phthalocyanines , Azo dyes , Indium compounds , Photochemotherapy , Nanoparticles , Photodegradation , Pollutants , Water -- Purification
  • Language: English
  • Type: text , Thesis , Masters , MSc
  • Identifier: http://hdl.handle.net/10962/67581 , vital:29116
  • Description: Indium (III) octacarboxyl phthalocyanine (ClInOCPc) alone and when conjugated to magnetic nanoparticles (MNP-ClInOCPc), 2(3),9(10),16(17),23(24)-octapyridylsulfanyl phthalocyaninato chloroindium (III) (ClInOPyPc) and its quaternized derivative 2(3),9(10),16(17),23(24)-octamethylpyridylsulfanyl phthalocyaninato chloroindium (III) (ClInOMePyPc) were synthesized. All Pcs were tested for both photodynamic antimicrobial chemotherapy (PACT) of an unknown water sample and photo-degradation of methyl red (MR). The singlet quantum yield (ΦΔ) for the ClInOCPc and MNP-ClInOCPc in PAN polymer fibers were 0.36 and 0.20 respectively using ADMA as a quencher in water. The photo-inactivation of bacteria in a water sample with unknown microbes was tested, with the MNP-ClInOCPc inactivating 90.6 % of the microbes and the ClInOCPc with 84.8 %. When embedded to the polymer, there was 48% bacterial clearance for ClInOCPc and 64% clearance for the MNP-ClInOCPc. The rate of degradation of MR increased with decrease of the MR concentration, with the MNP-ClInOCPc having the fastest rate. For ClInOPyPc and ClInOMePyPc, the singlet quantum yields were 0.46 and 0.33 in dimethylformamide (DMF), respectively. The PACT activity of ClInOMePyPc (containing 8 positive charges) was compared to those of 9(10),16(17),23(24)-tri-N-methyl-4-pyridylsulfanyl-2(3)-(4-aminophenoxy) phthalocyaninato chloro indium (III) triiodide (1) (containing 3 positive charges) and 2-[4-(N-methylpyridyloxy) phthalocyaninato] chloroindium (III) iodide (2) (containing 4 positive charges). When comparing ClInOMePyPc, 1 and 2, the largest log reduction for E. coli were obtained for complex 2 containing four positive charges hence showing it is not always the charge that determines the PACT activity, but the bridging atom in the phthalocyanine plays a role.
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  • Date Issued: 2019

Water quality, biomass and extracellular polymeric substances in an integrated algae pond system

  • Authors: Jimoh, Taobat Adekilekun
  • Date: 2018
  • Subjects: Water -- Purification , Sewage -- Purification -- Anaerobic treatment , Sewage lagoons , Sewage disposal plants , ASPAM model (Acid mine drainage) , Integrated algae pond systems (IAPS)
  • Language: English
  • Type: text , Thesis , Masters , MSc
  • Identifier: http://hdl.handle.net/10962/57307 , vital:26871
  • Description: Integrated algae pond systems (IAPS) combine the use of anaerobic and aerobic bioprocesses to effect wastewater treatment. Although, IAPS as a technology process offers many advantages including efficient and simultaneous N and P removal, no requirement for additional chemicals, O2 generation, CO2 mitigation, and a biomass with potential for valorization, a lack of technological advancement and the need for large land area, has limited the reach of this technology at industrial scale. In mitigation, peroxonation was introduced as a tertiary treatment unit and its effect on COD and TSS of IAPS treated water investigated. An effort was made to characterize the soluble but persistent COD in IAPS treated water and, productivity of the HRAOP mixed liquor was investigated to gain insight into the potential use of this biomass. Results show that peroxone treatment effectively reduced COD, TSS, and nutrient load of IAPS water without any significant impact on land area requirement. Indeed, summary data describing the effect of peroxone on quality of IAPS-treated water confirmed that it complies with the general limit values for either irrigation or discharge into a water resource that is not a listed water resource for volumes up to 2 ML of treated wastewater on any given day. Extraction followed by FT-IR spectroscopy was used to confirm albeit tentatively, the identity of the soluble but persistent COD in IAPS treated water as MaB-floc EPS. Results show that MaB-flocs from HRAOPs are assemblages of microorganisms produced as discrete aggregates as a result of microbial EPS production. A relationship between photosynthesis and EPS production was established by quantification of the EPS following exposure of MaB-flocs to either continuous light or darkness. Several novel strains of bacteria were isolated from HRAOP mixed liquor and 16S ribosomal genomic sequence analysis resulted in the molecular characterization of Planococcus maitriensis strain ECCN 45b. This is the first report of Planococcus maitriensis from a wastewater treatment process. Productivity and change in MaB-flocs concentration, measured as mixed liquor suspended solids (MLSS) between morning and evening were monitored and revealed that MLSS is composed of microalgae and bacteria but not fungi. Concentration varied from 77 mg L-1 in September (winter) to 285 mg L-1 in November (spring); pond productivity increased from 5.8 g m-2 d-1 (winter) to 21.5 g m-2 d-1 (spring); and, irrespective of MLSS concentration in late afternoon, approximately 39% was lost overnight, which presumably occurred due to passive removal by the algae settling pond. The outcomes of this research are discussed in terms of the quality of treated water, and the further development of IAPS as a platform technology for establishing a biorefinery within the wastewater treatment sector.
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  • Date Issued: 2018

Assessment of the prevalence of faecal coliforms and Escherichia coli o157:h7 in the final effluents of two wastewater treatment plants in Amahlathi Local Municipality of Eastern Cape Province, South Africa

  • Authors: Ajibade, Adefisoye Martins
  • Date: 2014
  • Subjects: Sewage disposal plants , Escherichia coli -- South Africa -- Eastern Cape , Escherichia coli O157:H7 , Escherichia coli , Effluent quality -- Testing , Whole effluent toxicity testing , Water -- Purification
  • Language: English
  • Type: Thesis , Masters , MSc (Microbiology)
  • Identifier: vital:11283 , http://hdl.handle.net/10353/d1016166 , Sewage disposal plants , Escherichia coli -- South Africa -- Eastern Cape , Escherichia coli O157:H7 , Escherichia coli , Effluent quality -- Testing , Whole effluent toxicity testing , Water -- Purification
  • Description: The production of final effluents that meet discharged requirements and guidelines remain a major challenge particularly in the developing world with the resultant problem of surface water pollution. This study assessed the physicochemical and microbiological qualities of two wastewater treatment works in the Eastern Cape Province of South Africa in terms of the prevalence of faecal coliforms and Escherichia coli O157:H7 over a five month period. All physicochemical and microbiological analyses were carried out using standard methods. Data were collected in triplicates and analysed statistically using IBM SPSS version 20.0. The ranges of some of the physicochemical parameters that complied with set guidelines include pH (6.7 – 7.6), TDS (107 – 171 mg/L), EC (168 – 266 μS/cm), Temperature (15 – 24oC), NO3- (0 – 8.2 mg/L), NO2- (0.14 – 0.71 mg/L) and PO4 (1.05 – 4.50 mg/L). Others including Turbidity (2.64 – 58.00 NTU), Free Cl (0.13 – 0.65 mg/L), DO (2.20 – 8.48 mg/L), BOD (0.13 – 6.85 mg/L) and COD (40 – 482 mg/L) did not comply with set guidelines. The microbiological parameters ranged 0 – 2.7 × 104 CFU/100 ml for FC and 0 – 9.3 × 103 for EHEC CFU/100 ml, an indication of non-compliance with set guidelines. Preliminary identification of 40 randomly selected presumptive enterohemorrhagic E. coli isolates by Gram’s staining and oxidase test shows 100% (all 40 selected isolates) to be Gram positive while 90% (36 randomly selected isolates) were oxidase negative. Statistical correlation between the physicochemical and the microbiological parameters were generally weak except in the case of free chlorine and DO where they showed inverse correlation with the microbiological parameters. The recovery of EHEC showed the inefficiency of the treatment processes to effectively inactivate the bacteria, and possibly other pathogenic bacteria that may be present in the treated wastewater. The assessment suggested the need for proper monitoring and a review of the treatment procedures used at these treatment works.
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  • Date Issued: 2014

Evaluation of some wastewater treatment facilities in Chris Hani and Amathole district municipalities as potential sources of Escherichia coli in the environment

  • Authors: Mazwi, Sinazo Nomathamsanqa
  • Date: 2014
  • Subjects: Escherichia coli -- South Africa -- Eastern Cape , Water -- Purification
  • Language: English
  • Type: Thesis , Masters , MSc (Microbiology)
  • Identifier: vital:11285 , http://hdl.handle.net/10353/d1019804 , Escherichia coli -- South Africa -- Eastern Cape , Water -- Purification
  • Description: Access to clean and safe water is essential for the survival of human beings. Pollution of freshwater sources constitutes a major problem hindering access to safe water for drinking and other domestic uses. Wastewater effluent discharges often impact the microbiological qualities of surface waters with its attendant health and environmental problems. This study evaluated the microbiological qualities of the discharged effluents of four selected wastewater treatment plants in Amathole and Chris Hani District Municipalities of the Eastern Cape Province over a twelve-month sampling period. Microbiological analysis (faecal coliform, Escherichia coli and Escherichia coli O157:H7) was done using standard methods and polymerase chain reaction method was used to confirm identities ofbacterial isolates. Presumptive bacteria counts ranged as follows: faecal coliforms 0 to 1.6 × 103 CFU/100 ml, E. coli 0 to 1.4 × 103 CFU/100 ml and E. coli O157:H7 0 to 9.6 × 102 CFU/100 ml. Forty eight percent (305/626) of the presumptive E. coli isolates were confirmed using species-specific uidA gene which code for β-glucuronidase enzyme in E. coli. Antibiotic susceptibility profile of the isolate using a panel of 10 antibiotics shows 100% (150/150) resistance to antibiotics rifampicin and penicillin G while 49.3% (74/150) of the isolates and 46.7% (70/150) were susceptible to streptomycin and cefotaxime respectively. Multiple antibiotic resistance phenotypes (MARP) of the isolates showed resistance to two or more test antibiotics while the calculated multiple antibiotic resistance index (MARI) for the tested isolated is 0.49. The detection of potentially pathogenic E. coli in the final effluents suggestspotential danger to the receiving water bodies where the effluents are discharge. The high MARI valued obtained in this study indicates that the isolates are form environment where the tested antibiotics are being used and may further lead to the spread of multiple antibiotics resistance among other pathogens that may be present in the same environment.
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  • 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.
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  • Date Issued: 2014

Assessment of the physicochemical and microbiological qualities of Tyume River in Amathole District in the Eastern Cape Province, South Africa

  • Authors: Sibanda, Timothy https://orcid.org/0000-0002-6864-3796
  • Date: 2013-05
  • Subjects: Water -- Purification , Sewage -- Purification
  • Language: English
  • Type: Doctoral theses , text
  • Identifier: http://hdl.handle.net/10353/24465 , vital:62812
  • 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 Tyume River water samples were collected monthly, over a 12-month period starting from August 2010 and ending in July 2011, and transported on ice to the Applied and Environmental Microbiology Research Group (AEMREG) Laboratory at the University of Fort Hare, Alice for analyses within 6 h of collection. Electrical conductivity (EC), total dissolved solids (TDS), temperature, pH and dissolved oxygen (DO) of water samples were determined in situ using a multi-parameter ion-specific meter. Concentrations of orthophosphate and total nitrogen (nitrate + nitrite) were determined by standard photometric methods. Total coliforms (TC), faecal coliforms (FC) and enterococci were determined by the membrane filtration method. Viruses in water samples were concentrated using the adsorption-elution method, followed by extraction of viral nucleic acids and purification done using commercially available kits. The concentrations of human enteric viruses in the river-water samples were estimated using quantitative PCR. RNA viruses were quantified in a two-step protocol where RNA was first transcribed into cDNA in a separate reverse-transcription step. Adenovirus species and serotypes were simultaneously detected using serotype-specific multiplex PCR. Norovirus genogroups GI and GII were detected by semi-nested PCR. The risk of infection associated with recreational and domestic use of the water was also estimated. Biochemical oxygen demand (BOD) levels fell within the stipulated BOD guideline of 10 mg/ℓ for surface waters where full contact use is allowed and ≤ 30 mg/ℓ where public access is prohibited, restricted, or infrequent. DO concentrations generally ranged between 7.47 mg/ℓ and 10.42 mg/ℓ, well within the target water quality requirements. The temperature regime ranged between 6°C and 28°C and for most sampling sites, the temperature regimes were within the acceptable limit of no risk (≤ 25°C) for domestic water uses in South Africa. EC ranged between 47 μS/cm and 408 μS/cm well within the South African target water quality EC guideline of 700 μS/cm though it was observed to increase as the river flowed through settlements. The pH in the period beginning September 2010 through to January 2011 was consistently below pH 9, but from February 2011 to June 2011 the pH significantly increased to between pH 10 and pH 11 at most sampling sites. Unpolluted waters normally show a pH of between 6.5 and 8.5. Most of the pH values observed in this study lie between pH 8.5 and pH 10.8 levels which are not far off from the upper level guideline of pH 9.0 for domestic use. Turbidity ranged between 6 NTU and 281 and fell short of the target water quality range (0 NTU to 1 NTU) of no risk for domestic water uses in South Africa. Monthly TDS values and EC values showed direct proportionality. TDS concentrations at all sites fell within the acceptable guideline of 0 mg/ℓ to 450 mg/ℓ of TDS for domestic use. Nutrient profiles were as follows: nitrate (0.18 mg/ℓ to 4.21 mg/ℓ); nitrite (0.02 mg/ℓ to 2.35 mg/ℓ); and orthophosphate (0.06 mg/ℓ to 2.72 mg/ℓ). The bacteriological qualities of the water were poor, exceeding the guideline of 200 CFU/100 mℓ and 33 CFU/100 mℓ for FC and enterococci respectively, for recreational water. FC counts also exceeded the 1 000 CFU/100 mℓ guideline for water used in fresh produce irrigation. Generally, higher counts of TC, FC and enterococci were recorded at the sampling sites located at the lower reaches of the river compared to the upper reaches. Adenovirus was detected in 31percent of the river samples in concentrations ranging between 1.0×100 genome copies/ℓ and 8.49×104 genome copies/ℓ. Serotyping showed the presence of species C adenovirus serotypes 1, 2, 6 and 7, and species F adenovirus serotype 41. The prevalence of norovirus was 4percent while rotavirus was detected in 4percent of river samples in concentrations ranging between 9×100 genome copies/ℓ and 5.64×103 genome copies/ℓ. Hepatitis A virus was detected in 13percent of river samples in concentrations ranging between1.67×103 and 1.64×104 genome copies/ℓ while enteroviruses were not detected. Detection of enteric viruses was inversely correlated to temperature. Risk analysis showed that both hepatitis A virus and adenovirus presented significantly higher risk of infection values compared to rotavirus in the case of ingestion of 10 mℓ or 100 mℓ of water from Tyume River while enteroviruses did not present any significant risk of infection. Tyume River water samples also did not conform to the US Environmental Protection Agency (US EPA) bacterial criteria of 200 CFU/100 mℓ faecal coliforms and 33 CFU/100 mℓ enterococci for bathing waters. Whereas the physicochemical parameters showed that Tyume River water was relatively clean, the bacteriological water quality was poor. Most of the microbiological contamination observed in this study (especially FIBs) can be blamed on inadequate sanitary infrastructure as we observed that open defecation is commonplace in this catchment, which also serves as a conduit for effluent discharges from wastewater-treatment facilities. Enteric viruses were detected along the course of the river in a sporadic pattern, generally not related to natural hydrological cycles and so we conclude that the presence of enteric viruses in the river is suggestive of the dynamics of the same in the host population. Even though the proportion of infective viruses was estimated in this study, fact remains that there is considerable risk of infection posed by the use of raw surface water for either domestic or recreational use. This study further confirmed the lack of correlation between faecal indicator bacteria and enteric virus occurrence in environmental waters, showing that assaying for enteric viruses in environmental waters remains the best method for determining the health risks associated with the use of faecally contaminated water. , Thesis (PhD) -- Faculty of Science and Agriculture, 2013
  • Full Text:
  • Date Issued: 2013-05

Development of a visible light active, photo-catalytic and antimicrobial nanocomposite of titanium dioxide and silicon dioxide for water treatment

  • Authors: Mungondori, Henry Heroe
  • Date: 2012
  • Subjects: Titanium dioxide , Silica , Catalysis , Nanocomposites (Materials) , Water -- Purification
  • Language: English
  • Type: Thesis , Masters , MSc (Chemistry)
  • Identifier: vital:11335 , http://hdl.handle.net/10353/471 , Titanium dioxide , Silica , Catalysis , Nanocomposites (Materials) , Water -- Purification
  • Description: The aim of this study was to prepare composite materials based on titanium dioxide (TiO2) and silicon dioxide (SiO2), and to evaluate their photo-catalytic and antimicrobial properties. Carbon and nitrogen doped TiO2nano-particles were prepared via a sol gel synthesis, which is a simple hydrolysis and condensation technique. In situ doping was carried out using glucose and urea as carbon and nitrogen sources respectively. Doping increased the spectral response of titanium dioxide photo-catalyst, allowing it to utilise the visible region which is much wider than the UV region (about 40 % of the solar spectrum), thus making it a more efficient photo-catalyst. The carbon and nitrogen doped TiO2-SiO2nano-particles were immobilized on glass support material to allow for easy separation of the spent photo-catalyst after the photo-degradation process. Tetraethyl orthosilicate (TEOS) was employed as both a binder and precursor for silicon dioxide. A mixture of TiO2 and TEOS in a 1:1 ratio was allowed to polymerize on a glass support which had been treated with hydrofluoric acid to introduce OH groups. The prepared photo-catalytic material was characterized by FT-IR, XRD, DRS, TEM, EDX, and BET analyses. Carbon was found to be more effective as a dopant than nitrogen. It brought about a band gap reduction of 0.30 eV and a BET surface area of 95.4 m2g-1 on the photo-catalyst as compared to a gap reduction of 0.2 eV and surface area of 52.2 m2g-1 for nitrogen doped TiO2. On the other hand, introduction of SiO2 allowed utilization of visible light by the TiO2-SiO2 nano-composite leading to an improved rate of photo-degradation of both methyl orange and phenol red. However, the immobilization of TiO2 on support material made it less effective towards inactivation of E. coli ATCC 25922 bacterial cells when compared to powdered TiO2 which was able to inactivate about 98 % of the bacterial cells within an hour of treatment.
  • Full Text:
  • Date Issued: 2012

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.
  • Full Text:
  • Date Issued: 2012

Preparation and evaluation of Lignocellulose-Montmorillonite nanocomposites for the adsorption of some heavy metals and organic dyes from aqueous solution

  • Authors: Bunhu, Tavengwa
  • Date: 2011
  • Subjects: Lignocellulose , Lignocellulose -- Biodegradation , Water -- Purification , Adsorption , Separation (Technology) , Dyes and dyeing , Montmorillonite
  • Language: English
  • Type: Thesis , Doctoral , PhD (Chemistry)
  • Identifier: vital:11333 , http://hdl.handle.net/10353/535 , Lignocellulose , Lignocellulose -- Biodegradation , Water -- Purification , Adsorption , Separation (Technology) , Dyes and dyeing , Montmorillonite
  • Description: The need to reduce the cost of adsorption technology has led scientists to explore the use of many low cost adsorbents especially those from renewable resources. Lignocellulose and montmorillonite clay have been identified as potentially low cost and efficient adsorbent materials for the removal of toxic heavy metals and organic substances from contaminated water. Montmorillonite clay has good adsorption properties and the potential for ion exchange. Lignocellulose possesses many hydroxyl, carbonyl and phenyl groups and therefore, both montmorillonite and lignocellulose are good candidates for the development of effective and low cost adsorbents in water treatment and purification. The aim of this study was to prepare composite materials based on lignocellulose and montmorillonite clay and subsequently evaluate their efficacy as adsorbents for heavy metal species and organic pollutants in aqueous solution. It was also important to assess the adsorption properties of the modified individual (uncombined) lignocellulose and montmorillonite. Lignocellulose and sodium-exchanged montmorillonite (NaMMT) clay were each separately modified with methyl methacrylate (MMA), methacrylic acid (MAA) and methacryloxypropyl trimethoxysilane (MPS) and used as adsorbents for the removal of heavy metals and dyes from aqueous solution. The lignocellulose and NaMMT were modified with MMA, MAA and MPS through free radical graft polymerisation and/or condensation reactions. NaMMT was also modified through Al-pillaring to give AlpMMT. The materials were characterised by fourier transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), scanning electron microscopy (SEM) and small angle X-ray scattering (SAXS) and characterisation results showed that the modification of the montmorillonite with MAA, MMA and MPS was successful. The modified lignocellulose and montmorillonite materials were evaluated for the adsorption of heavy metal ions (Cd2+ and Pb2+) from aqueous solution by the batch method. The adsorption isotherms and kinetics of both Cd2+ and Pb2+ onto the NaMMT clay, AlpMMT and lignocellulose materials are presented. The Langmuir isotherm was found to be the best fit for the adsorption of both heavy metals onto all the adsorbents. AlpMMT showed very poor uptake for heavy metals (both Cd2+ and Pb2+). PMMAgMMT, PMAAgMMT, PMAAgLig and PMPSgLig showed improved adsorption for both heavy metals. The mechanism of heavy metal adsorption onto the adsorbents was best represented by the pseudo second-order kinetic model. PMPSgLig, NaMMT and AlpMMT showed relatively high adsorption capacities for methyl orange, while the adsorption of neutral red was comparable for almost all the adsorbents. Neither the Langmuir model nor the Freundlich model was found to v adequately describe the adsorption process of dyes onto all the adsorbents. The pseudo second-order model was found to be the best fit to describe the adsorption mechanism of both dyes onto all the adsorbents. The modification of lignocellulose and montmorillonite with suitable organic groups can potentially produce highly effective and efficient adsorbents for the removal of both heavy metals and dyes from contaminated water. Novel adsorbent composite materials based on lignocellulose and montmorillonite clay (NaMMT) were also prepared and evaluated for the removal of pollutants (dyes and heavy metals) from aqueous solution. The lignocellulose-montmorillonite composites were prepared by in situ intercalative polymerisation, using methyl methacrylate, methacrylic acid and methacryloxypropyl trimethoxysilane (MPS) as coupling agents. The composite materials were characterised by FTIR, TGA, TEM and SAXS. SAXS diffractograms showed intercalated nanocomposites of PMMAgLig-NaMMT and PMAAgLig-NaMMT, whereas PMPSgLig-NaMMT showed a phase-separated composite and the same results were confirmed by TEM. The lignocellulose-montmorillonite composites were assessed for their adsorption properties for heavy metal ions (Cd2+ and Pb2+) and dyes (methyl orange and neutral red) from aqueous solution. Among these composite materials, only PMAAgLig-NaMMT showed a marked increase in the uptake of both Cd2+ and Pb2+ relative to lignocellulose and montmorillonite when used independently. The adsorption data were fitted to the Langmuir and Freundlich isotherms, as well as to the pseudo first-order and pseudo second-order kinetic models. The data were best described by the Langmuir isotherm and the pseudo second-order kinetic model. On the adsorption of dyes, only PMPSgLig-NaMMT showed enhanced adsorption of methyl orange (MetO) compared with lignocellulose and montmorillonite separately. The enhanced adsorption was attributed to the synergistic adsorption due to the presence of MPS, lignocellulose and NaMMT. Competitive adsorption studies were carried out from binary mixtures of MetO and Cd2+ or Pb2+ in aqueous solution. The adsorption process of MetO onto the composite material was found to follow the Freundlich adsorption model, while the mechanism of adsorption followed both the pseudo first-order and pseudo second-order models. This particular composite can be used for the simultaneous adsorption of both heavy metals and organic dyes from contaminated water. The adsorption of neutral red to the composite materials was comparable and the pseudo second-order kinetic model best described the adsorption mechanism.
  • Full Text:
  • Date Issued: 2011

Preparation of photocatalytic TiO₂ nanoparticles immobilized on carbon nanofibres for water purification

  • Authors: Nyamukamba, Pardon
  • Date: 2011
  • Subjects: Water -- Purification , Titanium alloys -- Industrial applications , Titanium , Nanoparticles , Drinking water -- Purification , Drinking water -- Contamination
  • Language: English
  • Type: Thesis , Masters , MSc (Chemistry)
  • Identifier: vital:11330 , http://hdl.handle.net/10353/367 , Water -- Purification , Titanium alloys -- Industrial applications , Titanium , Nanoparticles , Drinking water -- Purification , Drinking water -- Contamination
  • Description: Titanium dioxide nanoparticles were prepared using the sol-gel process. The effect of temperature and precursor concentration on particle size was investigated. The optimum conditions were then used to prepare carbon and nitrogen doped titanium dioxide (TiO2) nanoparticles. Doping was done to reduce band gap of the nanoparticles in order to utilize visible light in the photocatalytic degradation of organic compounds. A significant shift of the absorption edge to a longer wavelength (lower energy) from 420 nm to 456 nm and 420 nm to 428 nm was observed for the carbon doped and nitrogen doped TiO2 respectively. In this study, the prepared TiO2 photocatalyst was immobilized on carbon nanofibres to allow isolation and reuse of catalyst. The photocatalytic activity of the catalyst was tested using methyl orange as a model pollutant and was based on the decolourization of the dye as it was degraded. The doped TiO2 exhibited higher photocatalytic activity than the undoped TiO2. The materials prepared were characterized by XRD, TEM, SEM, FT-IR, DSC and TGA while the doped TiO2 was characterized by XPS, ESR and Raman Spectroscopy.
  • Full Text:
  • Date Issued: 2011

Towards a sustainable bioprocess for the remediation of acid mine drainage

  • Authors: Mambo, Mutsa Prudence
  • Date: 2011
  • Subjects: Acid mine drainage , Algae culture , Reduction (Chemistry) , Hydrolysis , ASPAM model (Acid mine drainage) , Water -- Purification
  • Language: English
  • Type: Thesis , Masters , MSc
  • Identifier: vital:5955 , http://hdl.handle.net/10962/d1006167 , Acid mine drainage , Algae culture , Reduction (Chemistry) , Hydrolysis , ASPAM model (Acid mine drainage) , Water -- Purification
  • Description: Acid mine drainage is of growing concern for both developing and developed economies. Thus there is increasing pressure to develop alternative remediation strategies. Biological sulphidogenic mechanisms have long since been studied but, very few have been implemented on a large scale. Limitations are due to the inability to acquire a suitable, low cost, environmentally friendly, renewable carbon source. The present study investigated the use of an algae biomass generated by the HRAOP of an IAPS as a carbon source for the EBRU 00AB/06 SRB consortium. The algae biomass and consortium were utilized together to remediate simulated AMD. Remediation involved decreasing the sulphate and metal concentrations in solution and decreasing the acidity of a simulated AMD. Experiments were carried out to investigate the capability of the EBRU 00AB/06 SRB consortium for sulphate reduction and sulphide generation. The consortium produced colonies when grown under anaerobic conditions in Petri dishes containing modified lactate SRB medium. The SRB consortium reduced the sulphate concentration of modified Postgates medium B and generated sulphide. Further analysis of the EBRU 00AB/06 SRB consortium revealed that the consortium was minimally impacted at pH 5 and by sulphate and iron at 3 g.L-1 and 0.5 g.L-1 respectively. The EBRU 00AB/06 SRB consortium was exposed to Actinomycin D and Ethidium Bromide to determine whether transcription and translation of proteins was required for sulphate reduction. Results indicated that sulphide generation and sulphate reduction were inducible. Analysis of the algae biomass used in this study revealed the empirical formula C1.0H1.91N0.084S0.003O0.36 indicating a carbon source rich in the nutrients required to sustain microbial development. Light microscopy revealed that algae cell walls and in particular those of Pediastrum were susceptible to acid hydrolysis. Dinitrosalicylic acid, Nile red, Bradford and Ninhydrin assays were used to determine the reducing sugar, lipid, protein and amino acid content respectively, of the mixed algae biomass. Results showed that upon exposure of the biomass to simulated AMD at pH 1 and pH 3, the concentration of reducing sugars and amino acids in solution increased. Whereas levels of lipids remained unchanged while the protein concentration decreased, indicating that, upon exposure of algae biomass to AMD, simulated or otherwise, cells ruptured, proteins were hydrolyzed and polysaccharides were broken down to sugars which are immediately available for SRB utilization. Exposure of biomass to simulated AMD revealed further that the presence of algae biomass increased the pH of simulated AMD (pH 3) to pH 7.67 after 4 d. Likewise, the pH of simulated AMD at 1 increased to 1.77 after 2 d while pH of the neutral control increased to 8.1 after 4 d. A direct comparison between lactate and algae biomass revealed 94 % sulphate removal after 23 d in the presence of algae biomass while 82 % sulphate removal was measured in the presence of lactate. Thus the EBRU 00AB/06 SRB consortium successfully utilized algae biomass for sulphate reduction and sulphide generation. In another experiment to establish if the consortium could remediate simulated AMD (pH 5) containing 0.5 g.L-1 iron and 3 g.L-1 sulphate while utilizing an algae biomass as the carbon source no residual iron was detected after 14 d and by day 23, an 89.07 % reduction in sulphate was measured. The results of this investigation are discussed in terms of utilizing a readily available and renewable biomass in the form of microalgae produced in HRAOPs as an effective carbon source in the SRB catalysed remediation of AMD.
  • Full Text:
  • Date Issued: 2011

Assessment of the prevalence of virulent Eschericia coli strains in the final effluents of wastewater treatment plants in the Eastern Cape Province of South Africa

  • Authors: Osode, Augustina Nwabuje
  • Date: 2010
  • Subjects: Escherichia coli , Escherichia coli -- Genetics , Effluent quality -- Testing , Water -- Purification , Sewage disposal plants , Escherichia coli -- South Africa -- Eastern Cape , Whole effluent toxicity testing
  • Language: English
  • Type: Thesis , Doctoral , PhD (Microbiology)
  • Identifier: vital:11246 , http://hdl.handle.net/10353/d1001062 , Escherichia coli , Escherichia coli -- Genetics , Effluent quality -- Testing , Water -- Purification , Sewage disposal plants , Escherichia coli -- South Africa -- Eastern Cape , Whole effluent toxicity testing
  • Description: Escherichia coli (E. coli) is a common inhabitant of surface waters in the developed and developing worlds. The majority of E. coli cells present in water are not particularly pathogenic to humans; however, there are some present in small proportion that possess virulence genes that allow them to colonize the digestive tract. Pathogenic E. coli causes acute and chronic diarrheal diseases, especially among children in developing countries and in travelers in these locales. The present study, conducted between August 2007 and July 2008, investigated the prevalence and distribution of virulent E. coli strains as either free or attached cells in the final effluents of three wastewater treatment plants located in the Eastern Cape Province of South Africa and its impact on the physico-chemical quality of the receiving water body. The wastewater treatment plants are located in urban (East Bank Reclamation Works, East London), peri-urban (Dimbaza Sewage Treatment Works) and in rural area (Alice Sewage Treatment Works). The effluent quality of the treatment plants were acceptable with respect to pH (6.9-7.8), temperature (13.8-22.0 °C), dissolved oxygen (DO) (4.9-7.8 mg/L), salinity (0.12-0.17 psu), total dissolved solids (TDS) (119-162 mg/ L) and nitrite concentration (0.1-0.4 mg/l). The other xii physicochemical parameters that did not comply with regulated standards include the following: phosphate (0.1-4.0 mg/L); chemical oxygen demand (COD) (5-211 mg/L); electrical conductivity (EC) (237-325 μS/cm) and Turbidity (7.7-62.7 NTU). Results suggest that eutrophication is intensified in the vicinity of the effluent discharge points, where phosphate and nitrate were found in high concentrations. Presumptive E. coli was isolated from the effluent samples by culture-based methods and confirmed using Polymerase Chain Reaction (PCR) techniques. Antibiogram assay was also carried out using standard in vitro methods on Mueller Hinton agar. The viable counts of presumptive E. coli for the effluent samples associated with 180 μm plankton size ranged between 0 – 4.30 × 101 cfu/ml in Dimbaza, 0 – 3.88 × 101 cfu/ml in Alice and 0 – 8.00 × 101 cfu/ml in East London. In the 60 μm plankton size category E. coli densities ranged between 0 and 4.2 × 101 cfu/ml in Dimbaza, 0 and 2.13 × 101 cfu/ml in Alice and 0 and 8.75 × 101 cfu/ml in East London. Whereas in the 20 μm plankton size category presumptive E. coli density varied from 0 to 5.0 × 101 cfu/ml in Dimbaza, 0 to 3.75 × 101 cfu/ml in Alice and 0 to 9.0 × 101 cfu/ml in East London. The free-living presumptive E. coli density ranged between 0 and 3.13 × 101 cfu/ml in Dimbaza, between 0 and 8.0 × 101 cfu/ml in Alice and between 0 and 9.5 × 101 cfu/ml in East London. Molecular analysis successfully amplified target genes (fliCH7, rfbEO157, ial and aap) which are characteristic of pathogenic E. coli strains. The PCR assays using uidA-specific primer confirmed that a genetic region homologous in size to the E. coli uidA structural gene, including the regulatory region, was present in 3 of the E. coli isolates from Alice, 10 from Dimbaza and 8 from East London. Of the 3 E. coli isolates from Alice, 1 (33.3%) was positive for the fliCH7 genes and 3 was positive for rfbEO157 genes. Out of the 10 isolates from Dimbaza, 4 were xiii positive for fliCH7 genes, 6 were positive for the rfbEO157 genes and 1 was positive for the aap genes; and of the 8 isolates from East London, 1 was positive for fliCH7 genes, 2 were for the rfbEO157 genes, 6 were positive for the ial genes. Antimicrobial susceptibility profile revealed that all of the E. coli strains isolated from the effluent water samples were resistant (R) to linezolid, polymyxin B, penicillin G and sulfamethoxazole. The E. coli isolates from Dimbaza (9/10) and East London (8/8) respectively were resistant to erythromycin. All the isolates were found to be susceptible (S) to amikacin, ceftazidime, ciprofloxacin, colistin sulphate, ceftriaxone, cefotaxime, cefuroxime, ertapenem, gatifloxacin, gentamycin, imidazole, kanamycin, meropenem, moxifloxacin, neomycin, netilmicin, norfloxacin and tobramycin. The findings of this study revealed that the Alice wastewater treatment plant was the most efficient as it produced the final effluent with the least pathogenic E. coli followed by the Dimbaza wastewater treatment plant. In addition, the findings showed that the wastewater treatment plant effluents are a veritable source of pathogenic E. coli in the Eastern Cape Province watershed. We suggest that to maximize public health protection, treated wastewater effluent quality should be diligently monitored pursuant to ensuring high quality of final effluents.
  • Full Text:
  • Date Issued: 2010

Ammonia removal from water by ion exchange using South African and Zambian zeolite samples

  • Authors: Mwale, Monica
  • Date: 2000
  • Subjects: Ammonia -- Toxicology , Water -- Purification , Zeolites , Ion exchange
  • Language: English
  • Type: Thesis , Masters , MSc
  • Identifier: vital:5215 , http://hdl.handle.net/10962/d1005058 , Ammonia -- Toxicology , Water -- Purification , Zeolites , Ion exchange
  • Description: One problem of intensive fish culture systems is the progressive build-up of toxic wastes such as ammonia. The possibility of improving aquaculture water quality using two kinds of zeolite is discussed. Zeolites are alumino-silicates whose framework allows them to exchange cations. Ion exchange has been demonstrated to be competitive with other methods of ammonia removal due to the high selectivity for ammonia exhibited by zeolite materials. In this study an unknown Zambian zeolite (identified as laumontite by X-ray diffraction techniques) and Pratley clinoptilolite (a South African zeolite) were tested under laboratory conditions and in a fresh water recirculating system. Ammonia cation exchange capacities (CEC) and suitable application rates for efficient water treatment were determined using the batch and column ion exchange procedures. Estimated ammonia uptake, the most important criterion used to assess performance of zeolite filters was strongly influenced by zeolite type, particle size, pre-treatment, regeneration and ion exchange method used. Statistical analysis showed significant differences in average ammonia CEC values between clinoptilolite (14.94 mg g⁻¹) and laumontite (2.77 mg g⁻¹), with the former displaying a higher Na⁺ ® NH₄⁺ exchange rate especially in the early reaction stages. This difference accords with the higher purity of clinoptilolite, 47% as opposed to 4.7% for laumontite, which makes it a better zeolite for ammonium removal. CEC increased linearly as particle size of the clinoptilolite was reduced resulting in a linear regression model (y = 18.29 – 3.704 x; r² = 74%). Pre-treatment of clinoptilolite using 1N NaCl significantly improved the ammonia CEC of clinoptilolite. Overall performance of both the batch and column methods achieved after regeneration (18.3 mg g⁻¹) was 25% higher than the estimated CEC values (13.0 mg g⁻¹) for the unregenerated samples of clinoptilolite. Comparison of CEC estimates using Pratley clinoptilolite, showed that average batch CEC estimates were significantly lower than the column method estimates. The average ammonia CEC values estimated in a fresh water recirculating system (5.80 mg g⁻¹ and 4.12 mg g⁻¹ for the 0.7-1.0 and 1.0-1.4 mm particle sizes, respectively) were significantly lower than the column and batch estimates for the same particle sizes (P < 0.05). Some nitrite (NO₂) and nitrate (NO3) build up was experienced probably due to the growth of autotrophs in the filters. Mass balance of nitrogen (N) for the three treatments of the fish trial (0.7-1.0 mm, 1.0-1.4 mm and the control treatment that had no zeolite in the filter) indicated that less that 10% of the N was retained for growth. It was found that 60% of the NH₄-N present associated with the soluble N was available for absorption by the zeolite filter or biological nitrification and that a total of approximately 22% of NH₄-N available was absorbed by clinoptilolite. The results indicate that the rate of nitrification can be deductively estimated by allowing a zeolite filter to become a biological filter. It is concluded that water treatment by ion exchange using natural zeolites, provides a reliable and efficient method for ammonia removal and appears to be a viable supplementary water treatment method for fresh water systems.
  • Full Text:
  • Date Issued: 2000

Capillary membrane-immobilised polyphenol oxidase and the bioremediation of industrial phenolic effluent

  • Authors: Edwards, Wade
  • Date: 1999
  • Subjects: Membranes (Technology) , Effluent quality , Pollutants , Phenols , Water -- Purification
  • Language: English
  • Type: Thesis , Doctoral , PhD
  • Identifier: vital:4095 , http://hdl.handle.net/10962/d1008458
  • Description: Waste-generating industrialisation is intrinsically associated with population and economic proliferation. This places considerable emphasis on South Africa's water shortage due to the integral relationship between population growth rate and infrastructure development. Of the various types of industry-generated effluents, those containing organic pollutants such as phenols are generally difficult to remediate. Much work has been reported in the literature on the use of enzymes for the removal of phenols from these waste-streams but little application of this bioremediation approach has reached practical fruition. This study focuses on integrating and synergistically combining the advantages of enzyme-mediated dephenolisation of synthetic and industrial effluent with that of membrane teclmology. The ability of the enzyme polyphenol oxidase to convert phenol and a number of its derivatives to chemically reactive o-quinones has been reported extensively in the literature. These o-quinones can then physically be removed from solution using various precipitation or adsorption techniques. The enzyme is, however, plagued by a product-induced phenomenon known as suicide inactivation, which renders it inactive and thus limits its application as a bioremediation tool. Integrating membrane technology with the enzyme's catalytic ability by immobilising polyphenol oxidase onto polysulphone and poly(ether sulphone) capillary membranes enabled the physical removal of these inhibitory products from the micro-environment of the immobilised enzyme which therefore increased the phenol conversion capability of the immobilised biocatalyst. Under non-immobilised conditions it was found that when exposed to a mixture of various phenols the substrate preference of the enzyme is a function of the R-group. Under immobilised conditions, however, the substrate preference of the enzyme becomes a function of certain transport constraints imposed by the capillary membrane itself. Furthermore, by integrating a quinone-removal process in the enzyme-immobilised bioreactor configuration, a 21-fold increase in the amount of substrate converted per Unit enzyme was observed when compared to the conversion capacity of the inunobilised enzyme without the product removal step. Comparisons were also made using different membrane bioreactor configurations (orientating the capillaries transverse as opposed to parallel to the module axis) and different immobilisation matrices (poly(ether sulphone) and polysulphone capillary membranes). Conversion efficiencies as high as 77% were maintained for several hours using the combination of transverse-flow modules and novel polysulphone capillary membranes. It was therefore concluded that immobilisation of polyphenol oxidase on capillary membranes does indeed show considerable potential for future development.
  • Full Text:
  • Date Issued: 1999

Evaluation of a 'defouling on demand' strategy for the ultrafiltration of brown water using activatable enzymes

  • Authors: Buchanan, K
  • Date: 1999
  • Subjects: Water -- Purification , Ultrafiltration , Enzymes , Membranes (Technology)
  • Language: English
  • Type: Thesis , Masters , MSc
  • Identifier: vital:3904 , http://hdl.handle.net/10962/d1003963 , Water -- Purification , Ultrafiltration , Enzymes , Membranes (Technology)
  • Description: New approaches to the application of membranes for the production of potable water are constantly being sought after in anticipation of future demands for increasingly rigorous water quality standards and reduced environmental impact. A major limitation, however, is membrane fouling, which manifests itself as a continual reduction in flux over time and thus restricts the practical implementation to restore flux. Mechanical and chemical methods have been implemented to restore flux to ultrafiltration systems, but these either result in a break in the process operation or lead to membrane damage or additional pollution problems. This project was aimed to develop a 'defouling on demand' stategy for cleaning membranes used during brown water ultrafiltration. The process involves the use of activatable peroxidase enzymes, which were immobilised onto flat sheet polysulphone membranes. Following flux decline which reaches a critical level with the build-up of the foulant layer, the immobilised enzyme layer was activated by the addition of a chemical activator solution, in this case hydrogen peroxidase and manganous sulphate. Manganese peroxidase was found to be the most effective enzyme at alleviating fouling by degrading the foulant layer formed on the membrane surface and hence restored flux to the ultrafiltration system. A 93% flux improvement was observed when manganese peroxidase was activated when 800uM manganous sulphate, 100mM hydrogen peroxide were added in the presence of a manganese chelator, lactate. The concept and the potential benefits this system holds will be discussed in further detail.
  • Full Text:
  • Date Issued: 1999

Analytical procedures for the determination of wattle polyphenols in wastewaters

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