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Investigation of Physical Characteristics Impacting Fate and Transport of Viral Surrogates in Water Systems Doctoral Dissertation

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Investigation of Physical Characteristics Impacting Fate and Transport of Viral Surrogates in Water Systems Doctoral Dissertation Investigation of Physical Characteristics Impacting Fate and Transport of Viral Surrogates in Water Systems Doctoral Dissertation Prepared by: Abigail J. Charest Department of Civil and Environmental Engineering Worcester Polytechnic Institute Worcester, MA Submitted to: Jeanine Plummer, Assoc. Professor, Civil & Environmental Engineering, Worcester Polytechnic Institute Sharon C. Long, Professor, Soil Science, University of Wisconsin – Madison John Bergendahl, Assoc. Professor, Civil & Environmental Engineering, Worcester Polytechnic Institute David Adams, Professor, Biology & Biotechnology, Worcester Polytechnic Institute Germano S. Iannacchione, Dept. Head, Physics, Worcester Polytechnic Institute Abstract A multi-scale approach was used to investigate the occurrence and physical characteristics of viral surrogates in water systems. This approach resulted in a methodology to quantify the dynamics and physical parameters of viral surrogates, including bacteriophages and nanoparticles. Physical parameters impacting the occurrence and survival of viruses can be incorporated into models that predict the levels of viral contamination in specific types of water. Multiple full-scale water systems (U.S., Italy and Australia) were tested including surface water, drinking water, stormwater and wastewater systems. Water quality parameters assessed included viral markers (TTV, polyomavirus, microviridae and adenovirus), bacteriophages (MS2 and ΦX-174), and coliforms (total coliforms and E. coli ). In this study, the lack of correlations between adenovirus and that of bacterial indicators suggests that these bacterial indicators are not suitable as indicators of viral contamination. In the wastewater samples, microviridae were correlated to the adenovirus, polyomavirus, and TTV. While TTV may have some qualities which are consistent with an indicator such as physical similarity to enteric viruses and occurrence in populations worldwide, the use of TTV as an indicator may be limited as a result of the detection occurrence. The limitations of TTV may impede further analysis and other markers such as coliphages, and microviridae may be easier to study in the near future. Batch scale adsorption tests were conducted. Protein-coated latex nanospheres were used to model bacteriophages (MS2 and ΦX-174) and includes a comparison of the zeta potentials in lab water, and two artificial groundwaters with monovalent and divalent electrolytes. This research shows that protein-coated particles have higher average log 10 removals than uncoated particles. Although, the method of fluorescently labeling nanoparticles may not provide consistent data at the nanoscale. The results show both that research on viruses at any scale can be difficult and that new methodologies are needed to analyze virus characteristics in water systems. A new dynamic light scattering methodology, area recorded generalized optical scattering (ARGOS) method, was developed for observing the dynamics of nanoparticles, including bacteriophages MS2 and ΦX-174. This method should be further utilized to predict virus fate and transport in environmental systems and through treatment processes. While the concentration of MS2 is higher than ΦX-174, as demonstrated by relative total intensity, the analysis of the variations of intensity over time shows that the dynamics are greater and have more variation in ΦX-174 than MS2 and this may be a result of the hydrophobic nature of ΦX-174. Relationships such as these should be further explored, and may reflect relationships such as particle bonds or hydrophobicity. ACKNOWLEDGEMENTS I am inexplicably grateful to a number of people and without their inspiration and guidance I would not have been able to complete this dissertation. First, I would like to thank my advisor, Jeanine Plummer. Her example has inspired me to be become a professor and without her support I never would have come back to graduate school to pursue my degree. I would like to thank Sharon Long who pushed me to take my research to the next level. I would like to thank John Bergendahl who fostered my interest in treatment processes. I would like to thank Dave Adams for igniting my love of viruses. I would like to thank German Iannacchione who invited me into his lab group, allowed me to be a part of a dynamic team and who provided me with the encouragement to expand my vision. Some faculty members of the Institute have been very kind enough to extend their help and support at various phases of this research, whenever I approached them, and I do hereby acknowledge them; Terri Camesano, Michael Johnson, Drew Brodeur, Susan Zhang, Richard Sisson, and Glenn Gaudette. I also want to thank my friends and lab partners Dan Roop, Evan Sullivan and Saad Algarni. I want to thank my dear friend, Linda Casill Vogel. I would not have been able to get to this stage without you. It all goes back to those midnight tutoring sessions in differential equations. I miss you. I am forever grateful to my family. This effort was not a solo act. It was only through their love, support and hard work that I was able to finish this degree. I want to thank my loving husband, Adrian Charest; he is the foundation that this degree is built upon. I want to thank my parents, John and Alison Thomas; they have loved me, encouraged me, inspired me and allowed me to grow into the person that I am. I want to thank my in-laws, George and Pauline Charest; their support has not only provided me with the encouragement to complete this degree, but their time and efforts have made this possible. Pauline Charest, I whole heartedly thank you for all the child care you provided during this process. You not only allowed me the time to pursue my degree, but you were Colette’s beloved companion. I always knew that she was not only safe, but happy and well loved. Finally, I want to thank my daughter, Colette Charest. This is for you. TABLE OF CONTENTS 1.0 Introduction .................................................................................................................................... 1-1 1.1 Viral Indicators in Full-Scale Water Systems ............................................................................ 1-2 1.2 Lab scale Analysis of Viral Surrogates ...................................................................................... 1-2 1.3 Nanoscale Analysis Utilizing Dynamic Light Scattering .......................................................... 1-3 2.0 Background .................................................................................................................................... 2-1 2.1 Traditional Bacterial Indicators ................................................................................................. 2-2 2.1.1 Coliform Bacteria ............................................................................................................ 2-3 2.1.2 Fecal Coliforms ................................................................................................................ 2-3 2.1.3 Escherichia coli ............................................................................................................... 2-3 2.1.4 Fecal Streptococci ............................................................................................................ 2-4 2.1.5 Methods of Detection ....................................................................................................... 2-4 2.2 United States Drinking Water Regulations ................................................................................ 2-6 2.2.1 Safe Drinking Water Act ................................................................................................. 2-7 2.2.2 Surface Water Treatment Rules ....................................................................................... 2-8 2.2.3 Groundwater Rule ............................................................................................................ 2-9 2.2.4 Total Coliform Rule ....................................................................................................... 2-10 2.3 Waterborne Disease Outbreaks ................................................................................................ 2-11 2.3.1 Surface Water Source .................................................................................................... 2-12 2.3.2 Groundwater Source ...................................................................................................... 2-13 2.3.3 Distribution System ....................................................................................................... 2-14 3.0 Indicator Systems for Viruses ........................................................................................................ 3-1 3.1 Indicator and Pathogen Alternatives .......................................................................................... 3-1 3.1.1 Bacteriophages ................................................................................................................. 3-2 3.1.1.1 Somatic Coliphages........................................................................................................ 3-4 3.1.1.2 Male-Specific Coliphages .............................................................................................. 3-4 3.1.1.3 Phages of Bacteroides Fragilis ....................................................................................... 3-4 3.1.2 Direct Monitoring of Viral Markers ...............................................................................
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