Enhanced biological phosphorus removal (EBPR) as well as biological nitrogen removal require a carbon source to be carried out. Volatile fatty acid (VFAs) (mainly acetic and propionic acids) are the major driving force for EBPR. Many domestic wastewaters have an insufficient amount of VFAs. However, carbon sources such as acetic and propionic acids can be produced using primary solids fermentation process. Due to the cost of VFA production, an external carbon source can be added to the biological nutrient removal (BNR) system that can be fermented to provide the desired VFAs. Glycerol (biodiesel by-product) offers a solution to reduce carbon addition cost if can be fermented to acetic and propionic acid or can be used directly as an external carbon substrate for EBPR and denitrification. Using glycerol in wastewater treatment can also offset the biodiesel plant disposal cost and reduce the BNR chemical cost. The main objective of this study was to optimize the prefermentation process and optimize the BNR system using glycerol as an external carbon source. In this work, Optimization of the prefermentation process using glycerol, mixing, and hydrogen gas addition was evaluated. EBPR performance within an A2O-BNR system was evaluated using either a direct glycerol method to the anaerobic zone or by co-fermentation with primary solids. Also, optimization of the nitrogen removal (specifically denitrification) efficiency of a 5-stage BardenphoTM BNR system using either a direct glycerol method to the second anoxic zone or by co-fermentation with primary solids was evaluated. It was found in this study that glycerol was an efficient external carbon substrate for EBPR as well as biological nitrogen removal. The prefermentation experiment showed that glycerol co-fermentation with primary solids produced significantly higher (p< 0.05) VFAs than primary solids fermentation alone, even more than the possible value from the added glycerol (427 mg-COD/L). The increased VFAs imply that the glycerol addition stimulated additional fermentation of primary solids. Lowering the prefermenter mixing energy (50 to 7 rpm) resulted in a significant increase in VFAs production (80%). Also, purging the headspace of the prefermenter with hydrogen gas did not lead to more VFAs, but significantly (p< 0.05) increased the propionic acid to acetic acid ratio by 41%. In the A2O-BNR pilot plant experiment, it was found that glycerol is a suitable renewable external substrate to drive enhanced EBPR as well as denitrification. The results from both locations of glycerol addition (direct vs. fermented) were beneficial to the BNR system. Both systems had similar effluent quality and achieved total nitrogen (TN) and total phosphorus (TP) removals up to 86% and 92% respectively. The 5-stage BardenphoTM BNR experiment investigated the location of glycerol addition (direct vs. fermented) on the performance of denitrification in the second anoxic zone and the overall performance. The results from both systems were that glycerol was beneficial to the BNR system and had virtually similar effluent quality. Both systems achieve complete denitrification and excellent removal of TN and TP up to 95% and 89% respectively. Also, the pilot that received fermented glycerol had significantly higher VFAs loading and lower observed yield. The side-stream prefermenter effluent flowing to the second anoxic reactor did not cause high effluent ammonia (NH3) concentration. In summary, the location at which glycerol was added did not affect effluent quality for nitrogen and phosphorus. However, glycerol addition and mixing energy did impact prefermenter performance and effluent quality.


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Graduation Date





Randall, Andrew


Doctor of Philosophy (Ph.D.)


College of Engineering and Computer Science


Civil, Environmental and Construction Engineering

Degree Program

Environmental Engineering









Release Date

August 2017

Length of Campus-only Access


Access Status

Doctoral Dissertation (Open Access)