UC Irvine UC Irvine Electronic Theses and Dissertations Title Understanding of Nitrifying and Denitrifying Bacterial Population Dynamics in an Activated Sludge Process Permalink https://escholarship.org/uc/item/1bd53495 Author Wang, Tongzhou Publication Date 2014 Peer reviewed|Thesis/dissertation eScholarship.org Powered by the California Digital Library University of California UNIVERSITY OF CALIFORNIA, IRVINE DISSERTATION Submitted in Partial Satisfaction of the Requirements for the Degree of DOCTOR OF PHILOSOPHY in Engineering by Tongzhou Wang Dissertation Committee Professor Betty H. Olson, Chair Professor Diego Rosso Professor Sunny C. Jiang 2014 © 2014 Tongzhou Wang ii To My Family ii Table of Contents Page LIST OF FIGURES .................................................................................................................. ix LIST OF TABLES .................................................................................................................... xi ACKNOWLEDGEMENTS ..................................................................................................... xii CURRICULUM VITAE ......................................................................................................... xiv ABSTRACT OF THE DISSERTATION ............................................................................... xvii Chapter 1. ................................................................................................................................ Introduction .................................................................................................................................................... 1 1.1 Background ...................................................................................................................... 1 1.2 Research Motivation ........................................................................................................ 4 1.3 Objectives and Hypothesis ............................................................................................... 6 1.4 Research Methodology .................................................................................................... 8 1.5 Organization of the dissertation ....................................................................................... 9 Chapter 2. ...................................................................................................................... Literature Review .................................................................................................................................................... 1 2.1 Nitrogen cycle ................................................................................................................ 11 2.2 Nitrification in Activated Sludge Process ...................................................................... 14 2.2.1 Nitrifiers .................................................................................................................. 14 2.3 Denitrification ................................................................................................................ 17 2.3.1 Denitrifying Enzyme ............................................................................................... 17 2.3.2 Dominant denitrifying populations in nitrifying-denitrifying activated sludge processes .......................................................................................................................... 18 2.4 Quantitative polymerase chain reaction (qPCR)............................................................ 21 Chapter 3. ... Understanding the role of Nitrifying Populations, Total Bacterial Populations and their effect on ammonia removal and denitrification in an Activated Sludge Process under Warm-Climate Condition ......................................................................................................... 23 3.1 Abstract .......................................................................................................................... 23 3.2 Introduction .................................................................................................................... 24 3.3 Materials and Methods ................................................................................................... 27 3.3.1 Plant Description and Sample Collection ............................................................... 27 3.3.2 DNA Extraction ...................................................................................................... 29 iii 3.3.3 Quantitative Polymerase Chain Reaction ............................................................... 29 3.3.4 Positive control for TB, AOB, NIS, and NB ........................................................... 31 3.3.5 Pearson’s correlation and principal component analysis ........................................ 31 3.4 Results ............................................................................................................................ 32 3.4.1 Biological and physicochemical parameters on overall ammonia removal and denitrification ................................................................................................................... 32 3.4.2 Determination of the most important physicochemical parameters affecting nitrifying bacteria using Pearson’s correlation ................................................................ 40 3.4.3 Principal component analysis ................................................................................. 42 3.4.4 The need for further research in finding denitrification indicators ......................... 47 3.5 Conclusions .................................................................................................................... 47 Chapter 4. .. Investigation on the effect of physicochemical parameters on denitrifying bacteria and denitrification process in a full-scale water reclamation plant .......................................... 49 4.1 Abstract .......................................................................................................................... 49 4.2 Introduction .................................................................................................................... 50 4.3 Materials and Methods ................................................................................................... 55 4.3.1 Plant Description and Sampling.............................................................................. 55 4.3.2 Sample Collection ................................................................................................... 55 4.3.3 DNA extraction ....................................................................................................... 55 4.3.4 Zoogloea- like bacteria spp. Culture ....................................................................... 56 4.3.5 Thauera- like bacteria Positive Control .................................................................. 56 4.3.6 qPCR Thauera-like bacteria and Zoogloea- Methyloversatilis-like bacteria Primers and Probes development .................................................................................................. 56 4.3.7 Quantitative Polymerase Chain Reaction (qPCR) .................................................. 57 4.3.8 Melt Curve .............................................................................................................. 60 4.3.9 PCR product purification, and DNA sequencing and Optimization of primer annealing temperature ...................................................................................................... 60 4.3.10 Statistical calculation ............................................................................................ 61 4.4 Results ............................................................................................................................ 62 4.4.1 Sequencing Results for Zoogloea-Methyloversatilis like bacteria and Thauera-like bacteria. ............................................................................................................................ 62 iv 4.4.2 Effect of Temperature on Zoogloea-Methyloversatilis-like bacteria and Thauera spp. bacteria communities ................................................................................................ 64 4.4.3 Relationship between substrate and Zoogloea- Methyloversatilis-like bacterial and Thauera-like bacterial populations in activated sludge ................................................... 67 4.4.4 Aerobic versus Anoxic Denitrification.................................................................... 71 4.6 Conclusions .................................................................................................................... 77 Chapter 5. .................................................. Denitrification, Competitions among Minor Organisms .................................................................................................................................................. 79 5.1 Abstract .......................................................................................................................... 79 5.2 Introduction .................................................................................................................... 80 5.3 Materials and Methods ................................................................................................... 83 5.3.1 Plant Description ....................................................................................................