Submitted in fulfillment of the requirements of the degree of Master of Applied Science in Biotechnology, Durban University of Technology, 2015. / In 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.

Includes list of publications issued during the candidature. / Bibliography: leaves 176-197. / iii, ix, 197, [22] leaves : ill., maps ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Phosphorus retention was measured in five pilot-scaled constructed wetlands with different configurations in terms of macrophyte species and substrates in order to evaluate the phosphorus removal efficiency of water plants and substrates in experimental ponds; assess the contribution of macrophytes to phosphorus removal through direct uptake, modification of water chemistry and impacts on the phosphorus adsorption characteristics of substrate; and, contribute to the optimal design and operation of constructed wetlands for the treatment of agricultural drainage water. / Thesis (Ph.D.)--University of Adelaide, Dept. of Soil and Water, 2002?