Faculty of Applied Sciences
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Item Impact of chemical oxygen demand to nitrogen ratio on anammox bacterial growth in an up-flow anaerobic sludge blanket reactor(2023) Msimango, Sandile Simiso; Kumari, Sheena; Nasr, Mahmoud; Bux, FaizalThe anaerobic oxidation of ammonium (ANAMMOX) process has been suggested as an economical and innovative means of removing nitrogen from wastewater. Nevertheless, very few studies have evaluated the effect of the chemical oxygen demand (COD) to nitrogen (N) ratio on bacterial communities in an ANAMMOX-mediated system. Heterotrophic bacteria can readily outcompete the slow-growing ANAMMOX bacteria in the presence of organic carbon. This study examined the effect of the organic carbon to nitrogen (C/N) ratio on the performance of ANAMMOX in an upflow sludge blanket reactor using synthetic wastewater as the feedstock. Two UASB reactors (UASB-A and UASB-B) were seeded with biomass from a labscale ANAMMOX reactor and operated for a period of 593 days. Both reactors were operated using similar operational conditions during the enrichment phase (0-400 days). Thereafter, the addition of organic carbon in the medium altered the C/N ratio of one of the reactors (UASBB). During this period, UASB-A served as a control reactor. A CN ratio of 1.0, 1.5, and 2.0 was achieved in the UASB B reactor by increasing the organic carbon concentration every 60 days. The reactors were analyzed at three-day intervals per week for nitrogen and COD removal efficiency. The quantitative PCR method was used to detect the dominant N-removing organisms within both reactors at different phases. In addition, cDNA quantification or reverse transcriptase qPCR (RT-qPCR) was also conducted to determine the dominant and active nitrifying communities. The results indicated that when the C/N ratio is 1.0, almost complete removal of NH4 + -N is observed (92%), and nitrogen removal efficiency (NRE) is approximately 82%. The ratios of ΔNO2 - /ΔNH4 + and ΔNO3 - /ΔNH4 + ratios during this phase (C/N=1) fluctuated from >1.25 to <1.6 and from >0.35 to <0.45 <0.11 to >1.6, respectively, which was within the range of the expected ANAMMOX stoichiometric ratio. In addition, when the C/N ratio was increased from 1 to 1.5, NRE rose from 82 to 88%. However, a decrease of NRE to 83% was observed when the C/N ratio was further increased to 2. The quantitative PCR results showed an increase in total bacteria from 1.4 × 106 copies/µL to 2.3× 106 copies/µL, and 2.4× 106 copies/µL as the ratio of C/N increased from 1.0 to 1.5 and thereafter to 2, respectively. ANAMMOX bacteria showed an increase from 16 × 103 copies/µL to 6.5× 10 4 copies/µL, and 2.06 × 105 copies/µL when the C/N ratio was increased from 1 to 1.5, and 2, respectively. The cDNA analysis further showed an increase of ANAMMOX bacteria transcript abundance from 4.6 × 104 copies/µL to 2.52× 106 copies/µL with an increase in C/N ratio to 1.5. Subsequently, a decrease in ANAMMOX bacteria transcript abundance to 1.09 × 106 copies/µL was observed when the C/N ratio was further increased to 2. The expression of the hzo gene encoding for hydrazine dehydrogenase (HDH), which catalyses the oxidization of the unique ANAMMOX intermediate hydrazine to N2 was 169 folds of expression, which was very high at C/N=1, but showed a decrease to 39 folds expression at C/N=1.5. Almost complete inhibition of hzo gene was observed when the C/N ratio was further increased to 2. Based on chemical analysis, it was further confirmed that the decrease of both ANAMMOX and AOB abundance at a higher C/N ratio caused an increase in effluent NH4 + -N concentrations. In conclusion, the study has shown that a higher C/N ratio could significantly affect the overall nitrogen removal rate and the activity of the diverse microbial populations, more specifically the ANAMMOX bacterial activity.Item An integrated approach for biofuel and fertilizer production using microalgae grown in wastewater(2022-09) Musetsho, Pfano; Bux, Faizal; Renuka, Nirmala; Guldhe, AbhishekMicroalgae are recognized as potential candidates for resource recovery from wastewater and are projected for biorefinery models. Therefore, this study was undertaken to evaluate the potential of poultry litter and municipal wastewater as nutrient and water sources, for the cultivation of Acutodesmus obliquus for lipids production for biodiesel application. The efficacy of lipid extracted biomass (LEA) as fertilizer for mung bean crops was also assessed in microcosm. A. obliquus cultivation in acid pre-treated poultry litter extract (PPLE) showed maximum biomass production of 1.90 g L-1 , which was 74.67% and 12.61% higher than the raw poultry litter extract (RPPE) and BG11 respectively. Higher NO3-N, NH3-N, and PO4-P removal of 79.51%, 81.82%, and 80.52% respectively were observed in PPLE as compared to RPLE treatment. The highest biomass (140.36 mg L-1 d -1 ), lipids (38.49 mg L-1 d -1 ), and carbohydrates (49.55 mg L-1 d -1 ) productivities were observed in the PPLE medium. The application of LEA as a fertilizer for mung bean crops showed improvement in plant growth and soil microbial activity. A maximum increase in organic carbon (59.5%) and dehydrogenase activity (130.8%) was observed in LEA amended soil which was significantly higher than chemical fertilizer (CF) control in 30 days. Whilst plant fresh weight and leaf chlorophyll in the LEA amended soil was comparable to whole algal biomass (WA) and CF control. The findings of the present study could be a basis for sustainable biorefinery for the valorization of wastewater for the production of microalgae-derived biofuel and byproducts for agricultural applications.Item Optimization of biomass and lipids production from microalgae using wastewater in a pilot scale raceway pond(2021) Rawat, Ismail; Bux, FaizalMicroalgae provide a sustainable renewable solution for the production of commodity products such as liquid biofuels. There are numerous benefits to using algae for the production of biofuels, however, the cost of production is a major hurdle to commercial-scale development. Major factors influencing the production of algae are the cost of nutrients, availability of water, contamination, and grazers. Research into algal biomass for biofuels production at laboratory scale does not translate directly to cultivation at large scale due to the change in cultivation conditions and the constant flux of environmental factors. This study focuses on the upstream processes of cultivation of biomass in a ~ 1146 m2 raceway pond. It demonstrates biomass productivity under different climatic conditions and utilisation of post-chlorinated wastewater as a water and nutrient source. The study further elucidates the population dynamics of the system and provides insight into the challenges faced during the cultivation of algae at large scale. An indigenous Scenedesmus sp. gave biomass productivity of 31.23 g/m2 /d with lipid production of 29.6 % lipid/g DCW in a 10 m2 raceway pond in a greenhouse using BG11. Biomass productivity was reduced to 13.09 g/m2 /d with a lipid content of 22.9 % lipid/g DCW under 3-fold higher irradiance. Biomass productivity of circular 3000L ponds at the large scale site resulted in the highest biomass and acceptable lipid content using 250mg/L NaNO3 although significantly lower than the 10 m2 raceway ponds. Wastewater has shown potential to replace conventional media. Post-chlorinated wastewater was found to have low levels of nitrogen and phosphorus but contained metals that act as micronutrients for algae. Supplemented wastewater proved to be an effective growth. Six individual runs of a covered 1146 m2 raceway pond driven by paddlewheel were conducted over 15 months. The average water temperature ranged from 20.61±0.68°C during mid-winter to 31.03±2.22°C in late summer. Daylight ranges from 10.25 to 14 hours in winter and summer respectively. The highest average light intensity was 359.00±212.71 µmol/m2 /s from Mid-winter to early spring and 645.44±330.58 µmol/m2 /s in late summer. Biomass productivities were low ranging from 2.7 to 7.34 g/m2 /d for most runs of the raceway pond, mainly due to the long periods of cultivation. Average productivity at day 7 for all raceway runs was 7.25 g/m2 /d. Adaptive Neuro-Fuzzy Inference System (ANFIS) modelling of the system elicited that the major factors affecting biomass productivity in the raceway pond were light intensity, pH, and depth for the raceway pond. The model showed that maximum biomass productivity is possible at a depth between 20 and 22 cm at light intensities between 200 and 400 µmol/m2 /s. pH in the range of 9 to 9.5 correlated positively with light intensity ranging from 200 to 1000 µmol/m2 /s with maximum biomass expected in the region of 400 to 500 µmol/m2 /s. The main algal constituents for the raceway ponds were Scenedesmus obliquus, Scenedesmus dimorphus, Chlorella, Keratococcus, and species of unidentified cyanobacteria. Either Scenedesmus or Chlorella was dominant for extended periods. Bacteria in open systems can have a positive or negative effect on the growth of microalgae but is dependent on the strains of microalgae and bacteria as well as prevailing conditions making these systems highly complex. Rhodobacteraceae, Plactomycetaceae, Xanthomonadaceae, Flavobacteriaceae, Phycisphaeraceae, Comamonadaceae, and Cyclobacteriaceae were found to be the major families of bacteria that proliferate at different levels during the cultivation period in the circular ponds and the raceway pond. These families of bacteria have several beneficial traits to algae cultivation however further investigation is required. Modelling the system revealed that pH, depth, and light intensity were factors having a substantial effect on biomass productivity. As the system was carbon limited addition of CO2 (preferably a waste stream) could significantly enhance the overall biomass productivity. A major factor negatively affecting biomass productivity was the size of the pond. Inadequate mixing impacts biomass productivity in terms of access to nutrients and gaseous exchange. Shorter periods of cultivation resulted in higher productivities. For the scale of the system, semi-continuous harvesting would be required to achieve shorter residence time. This must be balanced against the energy utilization and cost of harvesting potentially lower culture densitiesItem Solid-phase extraction of selected acidic pharmaceuticals from wastewater using a molecularly imprinted polymer(2017) Zunngu, Silindile Senamile; Mdluli, Phumlane S.; Madikizela, Lawrence Mzukisi; Chimuka, LukeIn this study, molecular modeling was used to investigate the intermolecular interactions between the functional monomer and ketoprofen which is an acidic pharmaceutical that possesses anti-inflammatory and analgesic activities. Ketoprofen is widely employed in medical care for treating musculoskeletal injury. This led to rational design of a molecularly imprinted polymer (MIP) that is selective to ketoprofen. Density functional theory (DFT) at B3LYP/6-31 level was used to investigate the intermolecular interaction between functional monomers and ketoprofen. Binding energy, ΔE, was used as an indication of the strength of the interaction that occurs between functional monomers and ketoprofen. 2-vinylpyridine (2-VP) as one of the functional monomers gave the lowest binding energy when compared to all the functional monomers investigated. Monomer-template interactions were further experimentally investigated using spectroscopic techniques such as Ultraviolet-visible and Fourier transform infrared (FTIR). A selective MIP for ketoprofen was synthesized using 2-vinylpyridine, ethylene glycol dimethacrylate, 1,1’-azobis(cyclohexanecarbonitrile), toluene/acetonitrile (9:1, v/v), and ketoprofen as a functional monomer, cross-linker, initiator, porogenic mixture, and template, respectively. The polymerization was performed at 60 °C for 16 h, and thereafter the temperature was increased to 80 °C for 24 h to achieve a solid monolith polymer. The non-imprinted polymer (NIP) was synthesized in a similar manner with the omission of ketoprofen. Characterization with thermogravimetric analysis (TGA) and powder X-ray diffraction (XRD) showed that the synthesized polymers were thermally stable and amorphous. Morphology of the particles were clearly visible, with MIP showing rough and irregular surface compared to NIP on the scanning electron microscopy (SEM). The characterization of the prominent functional groups on both MIP and NIP were performed using FTIR and nuclear magnetic resonance (NMR). The existence of hydroxyl was observed in the MIP; this was due to the presence of ketoprofen in the cavity. Prominent carbonyl group was an indication of the cross-linker present in both polymers. The synthesized MIP was applied as a selective sorbent in the solid-phase extraction of ketoprofen from the water. The extracted ketoprofen was monitored by high performance liquid chromatography (HPLC) coupled with UV/Vis detector. Several parameters were investigated for maximum recovery of ketoprofen from the spiked deionized water. The optimum method involved the conditioning of 14 mg MIP sorbent with 5 mL of methanol followed by equilibrating with 5 mL of deionized water adjusted to pH 2.5. Thereafter, 50 mL sample (pH 5) was loaded into the cartridge containing MIP sorbent followed by washing and eluting with 1% TEA/H2O and 100% methanol, respectively. Eluted compounds were quantified with HPLC. MIP was more selective to ketoprofen in the presence of other structural related competitors. The analytical method gave detection limits of 0.23, 0.17, and 0.09 mg L-1 in wastewater influent, effluent, and deionized water, respectively. The recovery for the wastewater influent and effluent spiked with 5 µg L-1 of ketoprofen was 68%, whereas 114% was obtained for deionized water. The concentrations of ketoprofen in the influent and effluent samples were in the ranges of 22.5 - 34.0 and 1.14 - 5.33 mg.L-1, respectively. The relative standard deviation (RSD) given as ± values indicates that the developed analytical method for the analysis of ketoprofen in wastewater was rapid, affordable, accurate, precise, sensitive, and selective.Item Development and assessment of a membrane bioreactor for wastewater treatment(2001) Osifo, Peter Ogbemudia; Bux, FaizalA woven fibre membrane unit was developed locally and used for this study. The membrane unit was submerged in the aerobic tank of an anoxic-aerobic biological treatment process to treat dairy effluent. Another unit similar to the process above was also set up but with a gravity settling tank replacing the membrane unit. Kinetic studies were done on both processes and their results compared. Effluent characteristics of both processes were also compared. For the membrane bioreactor, the effectiveness of two backflush methods ie air and water, were tested. In this study it was discovered that air backflush could remove the clogging formed at external and internal pores of the membrane better than water backflush measured in terms of rate of pressure recovery and energy consumption. The effluent quality of the membrane bioreactor (MBR) in terms of COD, nitrogen, suspended solids and turbidity was excellent under all conditions tested and was better than conventional activated sludge system. The COD removed based on the average value in MBR was 96.02 % compared to 94 % conventional system. A mass balance of the processes showed that 79 % in the form of nitrogen fed into both the MBR and conventional systems was denitrified. The effluent suspended solids from the MBR was not determinable while that of conventional system was 2.09 mglL. Maximum specific nitrification rate determined graphically was 3.0 d-I and 2.10 d-I in conventional and MBR respectively. The smaller value of nitrification rate in MBR could be attributed to large number of nitrifier volatile suspended solids in the system. The estimated true yield and decay rate coefficients of conventional system is 0.204 kgVSS/kgCOD and 0.013 d-I respectively. The flux obtained from this study is between 22 and 92 L/m2.h. However, the average flux value is 57 L/m2.h for the whole period of operation. Regular flux decline observed during operation was improved through regular backwash with air and chemical cleaning. The transmembrane pressure did not as whole increase more than 55 kPa due to frequent cleaning of the membrane surface.Item Identification of polyphosphate accumulating bacteria from pilot- and full scale nutrient removal activated sludges(1999) Atkinson, Blaise William; Bux, FaizalGeneral removal of phosphorus (P) from wastewater was introduced in Scandanavia in the late 1960's. At that time it was believed that P alone was limiting to algal growth and that the sole removal of P would solve the problem of eutrophication. However, we now know that both P and nitrogen (N) contribute to this deleterious effect and as such, much research has been conducted concerned with both the biological and chemical removal of these nutrients from sewage effluents. Enhanced biological phosphorus removal (EBPR), which is basically the biological accumulation of soluble P (as polyphosphate or poly-P) from the bulk liquid in excess of normal metabolic requirements, still tends to be sensitive to many external parameters and, as such, is subject to fluctuations. This makes it extremely difficult for wastewater treatment installations to achieve and maintain full compliance with strict discharge regulations. A more comprehensive understanding of the microbial community within the mixed liquor of a wastewater treatment system is therefore required which will ultimately assist in improving system design and performance. Chemical and civil engineers, when designing biological wastewater treatment systems, consider only the processes (biological or chemical) taking place within the reactor/s with little or no regard for the individual microbial species or the entire microbial community involved. Process design appears to be tackled empirically from a 'black box' approach; biological reactions or processes occurring within a system such as wastewater treatment are all lumped together and attributed to a single surrogate organism ie., the response of the surrogate to certain stimuli accounts for the total system response. This is similar to an analogy which Professor George Ekama (Dept of Civil Engineering, UCT), a leading scientist in wastewater treatment and process design, refers to where engineers, if, for example, are confronted with modelling the dynamics of carbon dioxide utilisation ofa forest, would recognise the accumulative system response and not give cognisance to each individual tree's contribution. It is true that if one had to consider every microbial species present in a highly organised community such as activated sludge, process models, designed to make quantitative and qualitative predictions as to the expected effluent quality from a particular design, would become increasingly complex and superfluous. It is evident from the countless accomplishments that engineers have succeeded, to a certain degree, in modelling wastewater treatment systems. One only has to consider the tremendous success of biological P (bio-P) removal and nitrification/denitrification processes at full-scale. However, there are limitations to this empirical approach and EBPR processes occasionally deteriorate in phosphate removal efficiency. In order to further optimise biological processes, whether they be organics oxidation, bio-P removal, nitrification or denitrification, biological community analyses will have to play a more significant role in design. The better microbial community structure and function is understood, the better the control and management of the system. With the advent of improved microbial identification and enumeration (to a certain extent) techniques (in situ), it was considered significant to investigate the mechanism ofbio-P removal and to elucidate which bacteria are actively responsible for this process. To this end, experimental work was conducted in two phases: \xAE laboratory, where samples of mixed liquor were obtained from a full-scale wastewater treatment facility exhibiting biological nutrient removal (BNR) characteristics and @ pilot plant, where an enhanced culture ofpolyphosphate accumulating organisms (PAO's) was developed and probed using molecular identification and enumeration techniques (as well as a cultivation-dependent approach). During phase \xAE of experimentatItem Extraction, characterisation and metal biosorption of extracellular polysaccharides from activated sludge(1998) Zondo, Raynold Mduduzi; Swalaha, Feroz MahomedWaste activated sludge is a biological adsorbent whose potential to remove metals from solution and effluent has been demonstrated. Extracellular polysaccharides (EPS) as components of activated sludge are thought to contribute to activated sludge metal biosorption. During the present study characterisation and determination of the metal biosorptive capabilities of domestic and industrial extracellular polysaccharides (EPS) revealed similarities both in terms of chemical composition and metal adsorption potential. Extracellular polysaccharides were extracted from activated sludge, obtained from domestic and industrial sludge treatment plants, using chemical techniques which involved sodium hydroxide extraction and solvent precipitation. A purification technique, which involved precipitation of protein with chloroform and removal of nucleic acids was developed. To assess the efficiency of the purification method, the ratio of extracted polysaccharide to the amount of protein present was determined. This provided an indication of the magnitude of EPS extracted in relation to the degree of cellular disruption. The type of activated sludge being treated was shown to be of particular importance. The quantity of EPS present in the original sample was found to be higher in domestic sludge than in industrial sludge. Purified EPS was fractionated in a column of DEAE-Sepharose CL-6B using stepwise pH gradient elution. Molecular weight distribution was conducted on a column of Sepharose CL-4B. Component monosaccharides were identified by paper chromatography. Monomers identified were glucose, fructose, glucuronic acid and galactosamine. Ion-exchange chromatography results demonstrated the presence of a number of different polysaccharide fractions while gel filtration results indicated a wide molecular weight distribution range of EPS from both domestic and industrial activated sludge. This indicated potential for variety in the EPS content of the activated sludge. Metal adsorption studies were conducted to determine the capabilities of EPS to adsorb metalsItem Evaluation of anaerobic sludges as metal biosorbents and development of a biotechnological process for metal ion removal from selected wastewater(1997) Bux, Faizal; Kasan, Hamanth C.As a result of rapid expansion of the industrial sector and increasing population, the environment has been under phenomenal stress. The volume of sewage and other effluents has increased tremendously in the last century. Globally, approximately .12 million tonnes of dry sludge biomass is produced and discarded of by landspreading, landfilling, incineration or dumping in lagoons and oceans. The discharge of industrial effluents into receiving waters has been documented to be the cause of severe environmental contamination. Heavy metals have been the cause of particular environmental concern. Their toxic and carcinogenic potentials at low concentrations, as well as the large quantities disposed to the environment, have prioritised them as leading contaminants. Current technologies of remediating heavy metal containing effluents are expensive and, in most cases, ineffective. Locally, most industries are merely diluting their effluents, thus resulting in the loss of valuable water resources. Waste sludges have shown the ability to adsorb heavy metals from their aqueous environment. Therefore, the current study attempted firstly, to compare biosorptive capacities of various waste sludges for a range of heavy metal ions, and secondly, to establish a relationship, if any, between biosorptive capacity and sludge surface charge. Finally, a laboratory scale biosorption process, encompassing desorption and recovery of metal ions from sludge surfaces, would have to be developed. Effluents used included pure, metal solutions of divalent zinc, cadmium, copper, nickel, trivalent and hexavalent chromium. In addition, synthetic effluents comprising a cocktail of the above-mentioned metal ions as well as an industrial effluent from a metal plating company were used. Five waste digested sludges were prepared and challenged against pure metal solutions to determine and compare their respective biosorptive capacities. Mechanisms of biosorption were elucidated using the Langmuir adsorption isotherm model. Sludge surface charge was determined using the millivolt quantification method. Upscaling of bioreactor trials to fully mixed laboratory scale was also investigated. These experiments encompassed the use of three sludges showing the greatest potential for biosorption and desorption using the selected mineral acid, H2S04, In addition, a simultaneous fully mixed biosorption and desorption process was designed and optimised. Subsequent trials involved comparing the latter process with a packed bed configuration whereby biomass was immobilised using poly sulfone resin. The overall comparative adsorptive capacities of the sludges (SI-SS) for metal ions in single solutions was S3 > S2 > S4 > SS > SI. Surface charge determination showed S3 to contain the most electronegative charge, with other sludges following in the same descending order as mentioned above. These findings supported the theory of a direct correlation between sludge surface charge and biosorptive potential. The affinity series of the sludges for metal ions followed the descending order of Cd2+ > Cu2+ > Ni2+ > Zn2+ > Cr6+ > Cr3+. Fully mixed studies, using mixed synthetic effluents, resulted in lower biosorptive capacities being recorded by the three selected sludges ie., S2, S3 and S4, as compared to single solution experiments. Biosorption studies with industrial effluent, containing Zn2+ as the most prevalent metal at 119.4 mg.F'. resulted in S3 biosorbing a maximum of 4.5 mg.g' of the cation. Sulphuric acid (H2S04) at O.2N, hydrochloric acid (HCI) at O.2N and acetic acid (CH3COOH) at O.4N were tested for their desorptive efficiencies. Sulphuric acid proved to be the most effective desorbing agent. Using S3 as biosorbent and O.2N H2S04 as desorbent, the manipulation and operation of a simultaneous process proved to be successful since both biosorption and desorption occurred concurrently, thus reducing time required for successful remediation considerably. Immobilised biomass, in a packed bed configuration, produced acceptable final effluent regarding standards as stipulated by the Durban Municipality for trade effluents. However, biosorption capacity of the sludge was compromised, with subsequent reductions in desorption being recorded, when the process was compared to fully mixed trials. Affinity series determined for the packed bed process wasC~+ >Cd2+>Zn2+>Cu2+>Cr6+ >Ni2+. Waste digested sludge has shown potential as metal biosorbent on an industrial scale. The present findings have succeeded in demonstrating a novel laboratory scale biotechnological process for the remediation of metal laden industrial effluents.Item Lab-scale assessment and adaptation of wastewater for cultivation of microalgal biomass for biodiesel production(2015) Ramanna, Luveshan; Bux, FaizalIn light of the world’s declining fossil fuel reserves, the use of microalgal biodiesel has come to the forefront as a potentially viable alternative liquid fuel. The depleting freshwater reserves make the feasibility of this concept questionable. The use of wastewater reduces the requirement for depleting freshwater supplies. This project aimed to determine the viability of municipal domestic wastewater effluent as a substrate for microalgal growth, in order to generate an economical and environmentally friendly source of biofuel. Wastewater effluents from three domestic wastewater treatment plants were characterized in terms of known microalgal nutrients viz., ammonia, phosphate and nitrates. Phosphate concentrations varied throughout the year and were found to be low (< 3 mgL-1) whilst ammonia and nitrate concentrations ranged from 0 to 10 mgL-1 throughout the experimental period. These wastewaters were found to be suitable for cultivating microalgae. The study explored the cultivation of Chlorella sorokiniana on pre- and post-chlorinated domestic wastewater effluent to assess their potential as a medium for high microalgal culture density and lipid production. Post-chlorinated wastewater effluent was found to be superior to pre-chlorinated wastewater effluent, as evident by the higher biomass concentration. This wastewater stream did not contain high concentrations of bacteria when compared to pre-chlorinated wastewater effluent. Nitrogen is an essential nutrient required for regulating the growth and lipid accumulation in microalgae. Cultures growing in post-chlorinated effluent had a lifespan of 18 d. Residual nitrogen in wastewater effluent supported microalgal growth for limited periods. Supplementation using cheap, readily available nitrogen sources was required for optimal biomass and lipid production. Urea, potassium nitrate, sodium nitrate and ammonium nitrate were evaluated in terms of biomass and lipid production of C. sorokiniana. Urea showed the highest biomass yield of 0.216 gL-1 and was selected for further experimentation. Urea concentrations (0–10 gL-1) were assessed for their effect on growth and microalgal physiology using pulse amplitude modulated fluorometry. A concentration of 1.5 gL-1 urea produced 0.218 gL-1 biomass and 61.52 % lipid by relative fluorescence. Physiological stress was evident by the decrease in relative Electron Transport Rate from 10.45 to 6.77 and quantum efficiency of photosystem II charge separation from 0.665 to 0.131. Gas chromatography analysis revealed that C16:0, C18:0, C18:1, C18:2 and C18:3 were the major fatty acids produced by C. sorokiniana. Wastewater effluent has been considered an important resource for economical and sustainable microalgal biomass/lipid production. The study showed that C. sorokiniana was sufficiently robust to be cultivated on wastewater effluent supplemented with urea. The results indicate that supplemented wastewater effluent was an acceptable alternative to conventional media. Using a relatively cheap nitrogen source like urea can certainly improve the techno-economics of large scale biodiesel production.Item Molecular analyses of pure cultures of filamentous bacteria isolated from activated sludge(2005) Naidoo, DashikaThe activated sludge process is the mostl used biological treatment process. Engineers and microbiologists are constantly seeking ways to improve process efficiency, which can be attributed to the increasing demand for fresh water supplies and proper environmental management. Since the inception of the activated sludge process, bulking and foaming have been major problems affecting its efficiency. Filamentous bacteria have been identified as the primary cause of bulking and foaming. Numerous attempts have been made to resolve this problem. Some of these attempts were effective as interim measures but failed as long term control strategies. The identification of filamentous bacteria and the study of their physiology have been hampered by the unreliability of conventional microbiological techniques. This is largely due to their morphological variations and inconsistent characteristics within different environments. To fully understand their role in promoting bulking and foaming, filamentous bacteria need to be characterized on a molecular level. The aim of this study was, therefore, to identify filamentous bacteria in pure culture with the purpose of validating these findings to the physiological traits of the pure cultures when they were isolated. Fourteen different filamentous cultures were used for this study. The cultures were identified using specific oligonucleotide probes via fluorescent in situ hybridisation and nucleotide sequencing. Prior to sequencing, an agarose gel and a denaturing gradient gel Electrophoresis profile were determined for each isolate. The various techniques were optimised specifically for the filamentous isolates. The isolates were identified as Gordonia amarae, Haliscomenobacter hydrossis, Acinetobacter sp./Type 1863, Type 021N, Thiothrix nivea, Sphaerotilus natans and Nocardioform organisms.