Lab-scale assessment and adaptation of wastewater for cultivation of microalgal biomass for biodiesel production
Authors:   , Ramanna Luveshan 
Published: August 2021

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.