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Investigating Fate of Silver Nanoparticles in Wastewater Biofilms Connie Marie Walden University of Arkansas, Fayetteville

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Investigating Fate of Silver Nanoparticles in Wastewater Biofilms Connie Marie Walden University of Arkansas, Fayetteville University of Arkansas, Fayetteville ScholarWorks@UARK Theses and Dissertations 12-2017 Investigating Fate of Silver Nanoparticles in Wastewater Biofilms Connie Marie Walden University of Arkansas, Fayetteville Follow this and additional works at: http://scholarworks.uark.edu/etd Part of the Civil Engineering Commons, and the Environmental Engineering Commons Recommended Citation Walden, Connie Marie, "Investigating Fate of Silver Nanoparticles in Wastewater Biofilms" (2017). Theses and Dissertations. 2549. http://scholarworks.uark.edu/etd/2549 This Dissertation is brought to you for free and open access by ScholarWorks@UARK. It has been accepted for inclusion in Theses and Dissertations by an authorized administrator of ScholarWorks@UARK. For more information, please contact [email protected], [email protected]. Investigating Fate of Silver Nanoparticles in Wastewater Biofilms A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Engineering by Connie Marie Walden University of Arkansas Bachelor of Science in Civil Engineering, 2012 University of Arkansas Master of Science in Civil Engineering, 2014 December 2017 University of Arkansas This dissertation is approved for recommendation to the Graduate Council. ____________________________ Dr. Wen Zhang Dissertation Director _____________________________ _____________________________ Dr. Julian Fairey Dr. Franck Carbonero Committee Member Committee Member _____________________________ Dr. Lauren Greenlee Committee Member Abstract As industrial advances make everyday life easier for human kind, the processes by which we need to maintain sanitary conditions for both water and wastewater treatment will become increasingly complex. Innovations in food packaging and textile design incorporate engineered nanoparticles (ENPs) to increase antimicrobial properties of clothing, maintain product color, and keep food in packaging from spoilage. For most products, ENPs released will enter the sanitary sewer system, and ultimately wastewater treatment plants. Biofilms grow universally on surfaces where a protective layer of extracellular polymeric substances (EPS) shields attached cells from stressors. In wastewater treatment, complex biofilms are utilized as a biological process for nutrient removal. Along with manufacturing innovations, the technology to study wastewater processes also continues to advance. Understanding complex biological communities requires detailed expertise in metagenomics for identifying bacteria present in a unit process of interest. This dissertation seeks to address both issues with respect to biofilm processes. First, a review of ENPs and their interaction with wastewater microbial communities lays groundwork for understanding the current state of knowledge. Then, a comparison of multiple methods to identify wastewater biofilms will help to understand the proper application of metagenomics to study changing biofilms in the presence of ENPs. Finally, multiple bench scale reactors and a quartz crystal microbalance are used to quantify ENP accumulation in wastewater biofilm. These studies advance the field of biofilm research by aiding in understanding how new technologies impact the biological treatment processes applied in wastewater treatment, as well as improve on the metagenomic identification of biofilm communities in these environments. Acknowledgments I would like to extend deepest gratitude to my advisor, Dr. Wen Zhang, for unwavering support and countless late-night revisions of each manuscript. I would not have been able to work this program and have a balanced home life without her understanding and encouragement. Besides my advisor, I would like to thank the rest of my advisory committee, Dr. Julian Fairey, Dr. Franck Carbonero, and Dr. Lauren Greenlee for providing thoughtful input throughout the years. I also acknowledge and am forever thankful for financial support provided by the University of Arkansas Graduate School Doctoral Academy Fellowship for the past four years, and the Hearst Foundation Fellowship through the Women in Engineering program. I am grateful to the other graduate students including Clint Mash, Johnnie Chamberlin and undergraduate Casey Gibson whom helped with gathering data, maintaining the laboratory, and sharing a laugh when things went terribly wrong. A special shout to Thien Do, whose insistence on a dozen training sessions on multiple devices not only made me chuckle, but also pull out my hair. Dedication This dissertation is dedicated to my parents for somehow molding me into the person I strive to be every day. And to my husband, thank you for riding this train to the end. Table of Contents Chapter 1: Introduction ..............................................................................................................1 1. Problem Statement .................................................................................................................2 2. Objectives and Approach .......................................................................................................3 2.1. Objective 1 ......................................................................................................................3 2.2. Objective 2 ......................................................................................................................3 2.3. Objective 3 ......................................................................................................................4 3. Document Organization .........................................................................................................4 4. References ..............................................................................................................................6 Chapter 2: Biofilms versus activated sludge: considerations in metal and metal oxide nanoparticle removal from wastewater ..............................................................................7 1. Introduction ............................................................................................................................8 1.1. Application and use of Me(O)NPs .................................................................................9 1.2. Presence of Me(O)NPs in wastewater ............................................................................11 2. Agglomeration/Aggregation of Me(O)NPs in wastewater ....................................................14 3. Interaction between Me(O)NPs and activated sludge. ...........................................................19 3.1. Removal of Me(O)NPs with activated sludge ................................................................19 3.2. Toxicity of Me(O)NPs and its impact on nutrient removal. ...........................................21 3.3. Impact on subsequent processes .....................................................................................23 4. Interactions between ENPs and Biofilm ................................................................................24 4.1. Entrapment of Me(O)NPs. ..............................................................................................26 4.2. Factors affecting removal of Me(O)NPs ........................................................................28 5. Me(O)NPs impact upon wastewater treatment design...........................................................30 5.1. Implications of Me(O)NPs in wastewater treatment processes……. .............................30 5.2. Considerations in regulating Me(O)NP removal from wastewater. ...............................33 Chapter 3: Assessing impacts of DNA extraction methods on next generation sequencing of water and wastewater samples ..................................................................................................... 55 1. Introduction. ...........................................................................................................................57 1.1 Materials and Methods.....................................................................................................59 1.1.1. Study sites and sampling........................................................................................59 1.1.2. Sample processing .................................................................................................59 1.1.3. DNA extractions ....................................................................................................60 1.1.3.1. QIAamp DNA Mini Kit. ...............................................................................60 1.1.3.2. QIAamp DNA Stool Mini Kit ......................................................................61 1.1.3.3. MO BIO PowerSoil DNA Kit. ......................................................................61 1.1.3.4. MO BIO PowerWater DNA Kit ...................................................................61 1.1.4. 16S rRNA amplification and Illumina sequencing. ...............................................61 2. Results and Discussion ..........................................................................................................62 2.1 Alpha diversity. ................................................................................................................63 2.1.1 Relative abundance. ...............................................................................................64 2.1.1.1 Freshwater lake. .............................................................................................64 2.1.1.2 Trickling filter
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