Dissertation submitted in compliance with the requirements for the Masters Degree in Technology: Biotechnology, Durban Institute of Technology, 2003. / Water is a precious and very valuable resource. Water resource shortages are problems that are plaguing the world. There is therefore a high demand for tightening water quality standards for both potable water and waters in the environment. It is becoming a necessity to treat and reuse wastewaters, especially wastewaters from industries, as these waters are of concern due to their toxic effects on the environment. In South Africa one of the industries of major concern is that of the edible oil industry and there are over a dozen ofthese industries in South Africa alone. Each of these industries utilises approximately 1.75 million m3 of water and discharges approximately 0.61 million m3 to the sewer each year. This wastewater that is being released has very high organic, inorganic and fats, oils and greases loads. Depending on the type of refinery process conducted on the crude oil, the effluent quality and quantity varies on a day to day basis. The two types of refinery procedures are the physical refining, where water is used or chemical refining where caustic soda is used. The organic load ofthe untreated effluent can range from 1 100 to 8 990 mg COD/L, the oils and greases can range from 80 to 1 360 mg/L and the pH can range from 1.8 to 10.5. / M

Dissertation submitted in compliance with the requirements for the Master's Degree in Technology: Biotechnology, Technikon Natal, 2001. / The vegetable oil industry produces effluent containing quantities of fat, oil, sodium, phosphates as well as other pollutants. Oils and greases tend to clog sewers and pumps, thus creating difficulties within the municipal wastewater treatment works. Physico-chemical treatment methods, such as (Dissolved Air Flotation) OAF, gravity separation and the use of coagulants have been attempted providing a considerable reduction in organic loading; however, discharge standards are still not met. Thus, biological treatment methods are being sought after. Aerobic treatment has been attempted however, shock loads cause problems while running such a process. The objective of this study was to assess the efficiency of anaerobic digestion to degrade Vegetable Oil Effluent (VOE) as well as the efficiency of the Anaerobic Baffled Reactor (ABR). Anaerobic digestion involves the breakdown of organic matter by the action of microorganisms in the absence of oxygen, producing methane-rich biogas. The VOE was characterized, providing significant information on its chemical composition. It was found that the effluent had high sulphate content as well as a high COD content. High sulpahte content of wastewaters have known to promote growth of Sulphate Reducing Bacteria (SRB), which utilize the same energy source as Methane Producing Bacteria (MPB) and therefore compete for the same energy source. Sulphate and lipid reduction pretreatment experiments were carried out, using barium chloride and gravitational separation respectively. The results obtained, showed that the use of barium chloride to reduce sulphate content in VOE was successful, with significant sulphate reduction. The lipid reduction experiments however, did not show any significant lipid reduction. Batch tests were conducted in serum bottles to assess the extent of biodegradation of the VOE in its raw state as well as with reduced sulpahte content. Methanogenic toxicity tests on the raw and pretreated VOE provided a range of toxicity results. These assays are relatively simple and inexpensive. Gas production was monitored to determine the rate and extent of biodegradation. The efficiency of digestion was assessed by COD reduction. Results indicated potential inhibition of the methanogenic bacteria responsible for methane production by the / M

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.