Thesis (Master of Engineering in Chemical Engineering)--Cape Peninsula University of Technology, 2017. / For more than a decade, poultry product consumption increased in developed and developing countries, with more than 470 new slaughterhouses being constructed in South Africa (SA). Customer demand for poultry products resulted in a rapidly growing poultry industry, with consequential increases in the quantity of organic solid and liquid waste being produced from the poultry slaughterhouses. Annually, the productivity and profitability within the livestock production sector has increased, an evaluation based on the number of slaughtered and sold animals. Potable water is required for these animals, resulting in the generation of high strength wastewaters. Instantaneous disposal of such wastewaters into the environment is concerning as it results in odour and the spreading of diseases in local rivers and freshwater sources. The generated poultry slaughterhouse wastewater (PSW) contains a high quantity of biodegradable organic, suspended and colloidal matter in the form of proteins, fats, oil and grease (FOG), protein from meat, blood, skin, and feathers, resulting in high Biological Oxygen Demand (BOD) and Chemical Oxygen Demand (COD), which can contribute to environmental deterioration if not treated adequately before discharge. On average, PSW contains a high concentration of BOD, COD, nitrogen, pathogenic and non-pathogenic viruses, bacteria and parasites, including their eggs. These characteristics make PSW highly polluted with a large quantity of bird carcass debris including FOG. Due to the high concentration of organic matter and suspended solids in the wastewater, it is necessary to pre-treat the PSW prior to sequential anaerobic treatment. Most of the contaminants present in the PSW can be reduced by means of numerous treatment steps, i.e. physical, chemical and biological treatment. For this study, biological treatment methods, physical separation methods, and a membrane bioreactor system, were used to treat PSW. The biological treatment methods used were an anaerobic digester (AD) followed by a single stage nitrification/denitrification reactor and then a third stage in which an ultrafiltration (UF) and Microfiltration (MF) membrane bioreactor (MBR) was used. The AD used was an Expanded Granular sludge Bed Reactor (EGSB) as anaerobic digestion is one of the most effective biological wastewater treatment methods used, as it reduces the organic matter to even produce biogas as a renewable energy source. The basis of anaerobic treatment method relies on suitable bacteria cultivated in the absence of dissolved oxygen, facilitating decomposition of organic matter into a renewable source such as biogas. Similarly, biological nitrification/denitrification processes for the removal of total nitrogen (TN) in wastewater has become one of the most commonly used processes within the wastewater treatment sector. Nitrification and denitrification processes can be performed by some microorganisms within the wastewater in Wastewater Treatment Plants (WWTPs) The PSW used was collected at different times from a local poultry slaughterhouse in the Western Cape (South Africa) and stored in a refrigerator at 4°C until it was fed to the first stage of the treatment which was the EGSB. Before being fed to the EGSB, the PSW was filtered with a sieve to remove feathers and agglomerated FOG to avoid clogging of the tubing. The EGSB was inoculated with 0.747 L anaerobic granular sludge, had a working volume of 2.7 L, an inner diameter of 0.065 m and a height of 0.872 m respectively. Ceramic marbles with an average diameter of 0.0157m were placed at the bottom of the bioreactor as packing for the underdrain and to maintain the granular sludge within the heated section of the bioreactor. The EGSB was fed with three types of PSW: 50% (v/v), 70% (v/v), which was diluted with distilled water. Thereafter once the system stabilised the reactor was fed with undiluted PSW (100%). Each dilution was operated at different Hydraulic Retention Times (HRTs) and Organic Loading Rates (OLRs), with average HRTs used being 62.5, 57.5 and 49.65 h. Furthermore, the average OLRs were 1, 2 and 3 g tCOD/L.day respectively. The performance of the EGSB was determined using tCOD, Total Suspended Solids (TSS) and FOG, with overall averaged removal rates for these constituents being 69%, 98% and 92% respectively. The highest tCOD removal of 93 % (optimal efficiency) was obtained at an average HRT of 57.5 h with a corresponding average OLR of 2 g tCOD/L.day.

Combined treatment technologies for the removal of waste carbon, nitrogen, and/or sulfur under anoxic/anaerobic conditions have recently received considerable attention. It has been reported that nitrate and/or reduced N-oxides, such as nitrite (NO2-), nitric oxide (NO), and nitrous oxide (N2O), which are products of denitrification, suppress methanogenesis. Research was conducted to investigate the effect of N-oxides and sulfide on mixed, mesophilic (35oC) methanogenic cultures, along with the effect of the type of electron donor on the kinetics and pathway of nitrate reduction. Among all N-oxides tested, NO exerted the most and nitrate exerted the least inhibitory effect on the fermentative/methanogenic consortia. Long-term exposure of a methanogenic culture to nitrate resulted in an increase of N-oxide reduction and a decrease of methane production rates. Sulfide addition to sulfide-free enriched cultures resulted in inhibition of NO2-, NO, and N2O reduction causing accumulation of these intermediates, which in turn inhibited methanogenesis and fermentation. In nitrate-amended, sulfide-acclimated cultures, nitrate reduction occurred via dissimilatory nitrate reduction to ammonia (DNRA); thus, accumulation of N-oxides was avoided and inhibition of methanogenesis was prevented. The nitrate reduction rates in cultures fed with different electron donors followed the descending order: H2/CO2 > acetate > glucose > dextrin/peptone > propionate. Denitrification was observed in the propionate-, acetate-, and H2/CO2-fed cultures regardless of the COD/N value. Both denitrification and DNRA were observed in the dextrin/peptone- and glucose-fed cultures and the predominance of either of the two pathways was a function of the COD/N value. Nitrate reduction processes were incorporated into the IWA Anaerobic Digestion Model No. 1 (ADM1) in order to account for the effect of nitrate reduction processes on fermentation and methanogenesis. The extended ADM1 described the experimental results very well. Model simulations showed that process interactions during nitrate reduction within an overall methanogenic system cannot be explained based on only stoichiometry and kinetics, especially for batch systems and/or continuous-flow systems with periodic, shock nitrate loads. The results of this research are useful in predicting the fate of carbon-, nitrogen-, and sulfur-bearing waste material, as well as in understanding microbial process interactions, in both natural and engineered anoxic/anaerobic systems.

This study evaluated physical and biological treatments of bio-oil process water to decrease organic contaminants. A three-sequential-column filtration system compared four treatments: three columns filled with kenaf only; three columns filled with wood shavings only; first column filled with wood shavings and two with kenaf; and first column filled with kenaf and two with wood shavings. The kenaf and wood shavings were composted after filtration. The filtrate water underwent further bio-treatment by adding aeration and selected bacteria. After filtration and bio-treatment, oil and grease concentrations were reduced over 80 percent and toxicity reduced over 90 percent. There were no significant differences among filtration treatments. Most of the oil and grease was removed by the first column. Aeration significantly decreased the concentration of oil and grease and toxicity in the filtrate water. Composting of the bioiltration matrices significantly reduced the oil and grease concentrations at day 45 by 80 percent.

The possibility of the existence of an iron-organic interaction in Harwood's Mill Reservoir contributing to a problem with floe formation after chlorinating filter-applied water was investigated. Shortened filtration-cycle times resulted when the filter-applied water contained the floc. The effects of varying pH, temperature, alum dosage, and oxidant addition on organic and meta.ls removals were examined with jar tests. Ultrafiltration analyses were performed to determine with which molecular size range of organic matter the iron may have been associated. Particle-size analysis was used to further examine the chlorination phenomenon. The low iron concentrations in the raw water were removed easily under any experimental condition. Organic removal, however, was optimized by alum coagulation ( 50 mg/L) at pH 5. 5 and a preoxidant dose of 2 mg/L. Improvements in organics removal over that of the WTP suggested that poor organic removal contributed to the floe-formation problem. / M.S.

[Truncated abstract] The use of constructed wetlands for tertiary purification of municipal wastewater has received increasing attention around the world because direct discharge of secondary-treated municipal wastewater to water bodies has caused eutrophication. Plant species selection and vegetation management may enhance nutrient removal efficiency in constructed wetlands. However, there is a lack of knowledge on the relations between plant growth and nutrient removal efficiency in constructed wetlands. The objective of this study is to better understand how plant growth and resource allocation are influenced by nutrients in wastewater and how nutrient removal efficiencies are affected by plant species and vegetation management. The preliminary experiment was conducted to select macrophytes, especially ornamental species, to grow in the wastewater in the wetland microcosms. Ten plant species, comprising six ornamental species: Alocasia macrorrhiza, Canna indica, Iris louisiana, Lythrum sp., Zantedeschia aethiopica, Zantedeschia sp., and four sedge species: Baumea articulate, Baumea juncea, Carex tereticaulis and Schoenoplectus validus, were planted in the wetland microcosms and fed a simulated wastewater solution in the concentrations similar to the secondary-treated municipal wastewater. C. indica has shown vigorous and healthy growth, and a relatively high potential of rooting-zone aeration and nutrient removal efficiency. B. articulata and S. validus also showed relatively high nutrient removal efficiency. ... The high nutrient availability and optimum N/P ratio were required for stimulating plant growth, resulting in allocation of more resources to above-ground tissues compared to below-ground parts, and enhancing nutrient removal efficiency. Nutrient removal efficiencies were significantly influenced by growth of C. indica and S. validus, nutrient loading rates and N/P ratios in the wastewater. The nutrient uptake kinetics of C. indica and S. validus were investigated to elucidate the differences in nutrient uptake between species. Wetland plant species have shown differential nutrient uptake efficiency and different preferences for inorganic N source, with C. indica preferring NO3-N and S. validus preferring NH4-N. C. indica had greater capacity than S. validus to take up PO4-P when the concentration of PO4-P in the solution was relatively low, whereas S. validus was more capable than C. indica to take up NO3-N when the concentration of NO3-N in the solution was relatively low. The PO4-P uptake capacity was higher in younger than older plants. Overall, the study has suggested that different plant species have differential capacity to take up nutrients. In addition to nutrient uptake, plants have significant other roles in terms of nutrient removal from the wastewater (such as leaking oxygen into the rhizosphere in which oxidation of substances like ammonia can occur). The properly high nutrient availability and optimum N/P ratio are required to stimulate the plant growth, resulting in enhancing the treatment performance in the wetlands. These findings have important implications for improving our ability to engineer ecological solutions to the problems associated with nutrient-rich wastewater.

The presence of heavy metal ions in waste streams is one of the most pervasive environmental issues of present times. A rotating biological contactor (RBC) was used to investigate the potential capacity of microbial biofilms in remediation of the metal ion species from a mixed metal contaminated effluent solution containing Cr+3 , Pb+2 and Cu+2 , each at a concentration of 200 mg r1 • In the first part of this study the effectiveness of various support materials for the development of microbial biofilms capable of removing heavy metals from a synthetic effluent was investigated. EDX analysis showed that none of the support matrices investigated, viz. gravel, polyester batting and sand, adsorbed metal ions on their surfaces; hence, metal adsorption was due purely to microbial activities. The biofilms attached more firmly and uniformly to polyester batting than to gravel and sand. The characteristics of polyester batting which made it a superior support matrix were its surface roughness and porous hydrophilic nature, which provided a larger surface area for the adhesion of microorganisms and attraction of nutrients during the biofilm development process. The selective accumulation of metal ion specIes by various microbial populations grown as biofilm using polyester batting as support matrix in separate compartments of a single-stage RBC bioreactor was examined. Lead ions were readily accumulated by almost all the microbial biofilms tested. Fungus-dominated biofilms selectively accumulated chromium ions whereas biofilms comprising mainly bacteria more readily accumulated copper ions from the mixed metal contaminated effluent solution. However, where interactions between the bacterial and fungal components were encouraged the mechanical stability of the biofilms was enhanced so that large amounts of all three metal ion species were removed by this biofilm. The combined effect of a series of bench-scale columns containing liquid humic acid and a three stage RBC bioreactor on the removal of metal ion species from a mixed metal contaminated effluent was investigated. After seven days of treatment the combined system had removed approximately 99% of the Cr+3, 98% of the Pb+2 and 90% of the Cu+2 ions from the mixed metal contaminated synthetic effluent. Complexation of the metal ions with humic acid was the predominant factor accounting for approximately 68-86% Cr+3 , 70-86% Pb+2 and 53-73% Cu+2 removal levels within the columns. A large proportion of the remaining Cr+3 and Pb+2, but not of the Cu+2, was removed in compartment 1 of the RBC. This suggested that the presence of the former two metals in solution might have reduced the removal of the Cu+2 ions from the system. The removal of substantially large amounts of the competing ions chromium and lead during the initial stages of the treatment process meant that copper was successfully taken up in the second and third RBC compartments. Hence, the economy of the treatment process was improved as larger quantities of the metal ions were removed in a shorter period of time than was possible when using the individual treatments (humic acid-metal complexation and biofilm adsorption) separately. More than 75%,92% and 86% of the adsorbed Cr+3 , Pb+2 and Cu+2 ions, respectively, were recovered from the three RBC bioreactor compartments following repeated washing of the biofilms with 0.1 M HCI. This relatively easy desorption suggested that the metal ions were simply adsorbed onto the surfaces of the biofilm cells rather than being taken into the cytoplasm of the cells. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2004.

Thesis (M.Tech.: Biotechnology)-Dept. of Biotechnolgy, Durban University of Technology, 2007 xviii, 143 leaves / Activated sludge flocs are responsible for flocculation, settling and dewaterability. It is important to maintain the growth off loc-forming bacteria for efficient sludge settleability and compaction for good quality effluent. Filamentous bacteria on the other hand are believed to provide rigid support network or backbone upon which floc-forming bacteria adhere to form stable activated sludge flocs (Wilderer et al., 2002; Ramothokang et al., 2003). Filamentous bacteria can also be detrimental to the process when they outgrow floc-forming bacteria. Morphologically filamentous bacteria are at an advantage as they have higher outward growth velocity and can extend freely to bulk liquid substrate. Proliferation of filamentous bacteria causes foaming and bulking (Martins et al., 2004). Although chemical alleviation measures to circumvent bulking are present, they are symptomatic (Chang et al., 2004). Eikelboom (1975) developed the first identification keys for the classification of filamentous bacteria that is primarily based on morphological characteristics and microscopic examination. Although very useful, this type of identification has its limitations. For instance some filamentous bacteria can change morphology in response to changes in the environment and although some of them can be morphologically similar they may vary considerably in their physiology and taxonomy (Martins et al., 2004). A vast number of filamentous bacteria are still very poorly understood which could be due to the problems of cultivation due to their slow growing nature and maintenance of cultures (Rossetti et al., 2006). This limitation necessitates a molecular approach to resolve the taxonomy of filamentous bacteria as it is a culture-independent technique which is highly accurate. This project was undertaken to verify the identity of pure cultures of filamentous bacteria isolated previously through the application of molecular techniques. The 16S rDNA are conserved regions in bacterial cells and they can be extracted and specific nucleic acid fragments amplified. Denaturation gradient gel electrophoresis enabled the separation of fragments of identical length but different size and served as an indication of purity (Muyzer et al., 1993).

Dissertation submitted in fulfilment with the requirements for the Masters Degree: Biotechnology, Durban University of Technology, 2011. / Untreated vegetable oil effluents (VOEs) are known for creating shock-loading problems for the receiving wastewater treatment installations, resulting in poor quality final effluents being produced which do not satisfy municipal discharge standards. Onsite activated sludge treatment as an alternative has not been fully investigated. Hence, in this investigation biological treatment using the activated sludge process was chosen as the method for the treatment of VOE. The effect of VOE on measured process parameters was also determined. Novel molecular techniques such as fluorescent in situ hybridisation (FISH) and dot-blot hybridization have become powerful tools for the analysis of complex microbial communities that exist within activated sludge. The aim of this investigation was to evaluate biological treatment, optimize and apply FISH and dot-blot hybridization in order to analyze the microbial community implicated the biological treatment of VOE using probes EUBmix, ALF1b, BET42a, GAM42a and HGC69a. A laboratory-scale modified Ludzack-Ettinger (MLE) process setup and fed VOE with a COD (chemical oxygen demand) of ± 1000 mg/L. Daily monitoring of the process involved COD and TKN (total kjeldahl nitrogen) analysis of the influent and effluent as well as direct OUR (oxygen utilization rate) measurement and monitoring of the MLVSS (mixed liquor volatile suspended solids) concentration of the aerobic mixed liquor. The process exhibited overall COD and TKN removal capacities of 84% and 90% respectively. The aerobic mixed liquor had an OUR of 19 mgO/L.h and an average MLVSS concentration of 3000 mg/L. FISH results revealed that 72% of cells stained with 4‟, 6-diamidino-2-phenylindole (DAPI) within the aerobic mixed liquor bound to probe EUBmix, indicating a substantial Bacterial population within the laboratory-scale biological process. The alpha-Proteobacteria was identified as the dominant bacterial community comprising 31% of Bacterial cells, followed by the beta-Proteobacteria (17% of EUBmix), gamma-Proteobacteria (8% of EUBmix) and Actinobacteria (4% of EUBmix). Results of dot-blot hybridization were in agreement with FISH Adrian Phillip Degenaar| CHAPTER 1: General Introduction - v - results reiterating dominance of the alpha-Proteobacteria. This indicated that the class alpha-Proteobacteria could play a primary role in the biological degradation of VOE. This research will therefore aid in process design and retrofitting of biological processes treating VOE.

M. Tech. (Chemical Engineering) / The usual target of an upgrading process using membrane is to produce a retentate stream, the product, with high CH4 concentration. This work presents a simulation of two possible membrane configurations, single stage without recycle (SSWR) and double stage with permeate recycle (DSPR), of an existing operational biogas upgrading plant. The simulation was conducted using ChemCAD and AlmeeSoft gas permeation software to investigate the performance of the configurations on product purity, recovery and required compressor power with a view to determine the optimal operational conditions for maximising the concentration of CH4 and its recovery. Thereafter, an economic assessment on the optimal configuration was conducted to determine the gas processing cost (GPC), the profitability of producing biomethane and cost-benefit of utilising biomethane as a vehicular fuel. The simulation was validated against plant data with a maximum percentage error of 2.64%. Increasing CO2 in feed reduced product recovery and purity. Increasing feed pressure and selectivity increased product recovery and purity up to the pressure limit of the membrane module. Increasing feed flow rate increased product recovery but reduces purity. In both configurations, increasing CO2 in the feed and increasing feed pressure increased the GPC. However, increasing feed flow rate reduced the GPC. The overall performance of DSPR configuration was much higher due to increased trans-membrane area available for separation. At optimal conditions, a product purity of 91% and 96% CH4 recovery was achieved from the initial plant result of 87.2% product purity and 91.16% CH4 recovery. The total compression duty was 141 kW. The GPC was $0.46/m3 of biomethane. The cumulative discounted NPV, IRR and BCR for producing biomethane was R15,240,343, 22.41% and 2.05 respectively, with a break-even in the 5th year after plant start-up considering a prime lending rate at 9%. Using CBG instead of gasoline saves 34% of annual fuel cost with a payback period of one year and three months for the cost of retrofitting the vehicle.

The purpose of this research is to hydraulically characterize an engineered wetland in Albany, Oregon. The wetland receives treated wastewater from both Albany Millersburg Water Reclamation Facility (AMWRF) and ATI Wah Chang. AMWRF's water is municipal waste water. ATI Wah Chang's water comes from its nearby metal processing plant. The wetland is designed to remove thermal input as well as nitrogen species from both sources. ATI Wah Chang effluent has significant nitrate concentrations. A reliable model is needed to estimate the denitrification potential of the wetland. In order to construct a model, accurate hydraulic parameters such as residence time and flow rate are needed. In the first few days after ATI started flow, the aquatic conductivity level of the wetland increased significantly. Conductivity was used as a tracer to estimate residence times in the wetland as well as to measure the split ratios from different water sources in the wetland (ATI Wah Chang and AMWRF). A pilot test on conductivity and flow rate was carried out on a single pond. The pilot test was designed to accurately measure the influent and effluent from a single pond. Using this information, rates of infiltration as well as unintended flow paths could be identified. A third tracer test was performed using Rhodamine W.T. This test allowed for the determination of the residence time of each pond, the wetland as a whole, and identified stagnant zones within the ponds. To simulate the nitrogen transportation and transformation process, a numerical model was developed. The model's input parameters include reaction rate constants for nitrification and denitrification, volume of each pond, flow rate, flow path connections, and temperature. The model simulated the tracer test that was performed on the wetland to verify its accuracy. The model is also capable of predicting denitrification potential in both pilot scale and field scale. It is also temperature sensitive because temperatures vary significantly; for instance, in winter when the average temperature in Albany is <5��C, denitrification rates decrease significantly. Through this research, hydraulic characterization as well as current denitrification rates in the wetland were identified. Strategies for increasing the denitrification rate were also identified through this research. / Graduation date: 2012