The enhanced biological phosphorus removal (EBPR) process efficiency relies on different operational and process conditions especially the type of carbon source available in the wastewater. Acetic acid and propionic acid are the two major volatile fatty acids (VFAs) found in domestic wastewater which can drive biological phosphorus (P) removal to the desired level. However, often domestic wastewater does not have a sufficient amount of VFAs. Due to high acetate and propionate production-cost, it is not economic to add acetate and propionate directly in full-scale wastewater treatment plants. This brought up the idea of using external carbon sources (e. g. molasses has been used successfully) in EBPR systems that can be converted to VFAs through a fermentation process. On the other hand, biodiesel fuels have been produced increasingly over the last decade. Crude glycerol is a biodiesel production major by-product that can be used as an external carbon source in wastewater treatment plant. Therefore, the main objective of this research is to optimize the glycerol/biodiesel waste fermentation process' operational conditions in pursuit of producing more favorable fermentation end-products (i. e. a mixture of acetic acid and propionic acid) by adding glycerol to a prefermenter versus direct addition to the anaerobic zone or fermentation with waste activated sludge. For this reason, different prefermenter parameters namely: mixing intensity, pH, temperature and solids retention time (SRT), were studied in a small scale fermentation media (serum bottles) and bench scale semi-continuous batch prefermenters. Experimental results revealed that glycerol/biodiesel waste fermentation resulted in a significant amount of VFAs production with propionic acid as the superior end-product followed by acetic acid and butyric acid. The VFA production was at its highest level when the initial pH was adjusted to 7 and 8.5. However, the optimum pH with respect to propionic acid production was 7. Increasing the temperature in serum bottles favored the total VFA production, specifically in the form of propionic acid. Regarding the mixing energy inconsistent results were obtained in the serum bottles compared to the bench scale prefermenters. The VFA production in mixed serum bottles at 200 rpm was higher than that of un-mixed ones. On the other hand, the unmixed or slowly mixed bench scale prefermenters showed higher VFA production than the mixed reactors. However, the serum bottles did not operate long enough to account for biomass acclimation and other long-term effects that the prefermenter experiments could account for. As a consequence one of the most important and consistently results was that VFA production was significantly enhanced by reducing mixing intensity from 100 rpm to 7 rpm and even ceasing mixing all together. This was true both for primary solids and glycerol. In addition propionate content was high under both high and low intensity, and adding glycerol also increased the fraction of primary solids that formed propionic acid instead of acetic acid. Increasing the SRT from 2 to 4 days increased the VFA production about 12% on average. In order to investigate the effect of glycerol on EBPR process efficiency two identical A2/O systems were monitored for 3 months. Experimental results suggested that glycerol addition could increase the P removal efficiency significantly. Adding glycerol to the prefermenter rather than the anaerobic zone resulted in a lower effluent soluble ortho phosphorus (SOP) (0.4 mg-P/L vs. 0.6 mg-P/L) but the difference was apparently statistically significant. Future experimentation should be done to determine if this effect is consistent, especially in carbon poor wastewaters. Also it would be desirable to conduct a longer pilot study or a full scale study to determine if this improvement in effluent SOP remains true over a range of temperature and changing influent conditions.


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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

June 2016

Length of Campus-only Access


Access Status

Doctoral Dissertation (Open Access)