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University of Oklahoma Graduate College Viral UNIVERSITY OF OKLAHOMA GRADUATE COLLEGE VIRAL METAGENOMICS AND ANTHROPOLOGY IN THE AMERICAS A DISSERTATION SUBMITTED TO THE GRADUATE FACULTY in partial fulfillment of the requirements for the Degree of DOCTOR OF PHILOSOPHY By ANDREW T. OZGA Norman, Oklahoma 2015 VIRAL METAGENOMICS AND ANTHROPOLOGY IN THE AMERICAS A DISSERTATION APPROVED FOR THE DEPARTMENT OF ANTHROPOLOGY BY ______________________________ Dr. Cecil M. Lewis, Jr., Chair ______________________________ Dr. Katherine Hirschfeld ______________________________ Dr. Paul Spicer ______________________________ Dr. Kermyt G. Anderson ______________________________ Dr. Tyrrell Conway © Copyright by ANDREW T. OZGA 2015 All Rights Reserved. Acknowledgements This dissertation would not have been possible without the guidance and encouragement of a number of professors, colleagues, friends, and family members. First and foremost I’d like to thank my advisor and chair, Dr. Cecil Lewis. His guidance and critiques throughout the project design, execution, and drafting process were invaluable to my completion of this degree. I’d also like to thank my doctoral committee: Dr. Tassie Hirschfeld, Dr. KG Anderson, Dr. Paul Spicer, and Dr. Tyrrell Conway for their support and insight regarding my project design and manuscript draft. I must thank those that graciously provided fecal samples for this project including the Cheyenne & Arapaho Nation, University of Oklahoma colleagues and students, and those from the coastal and jungle regions of Peru. Additionally, I’d like to thank my LMAMR colleagues including Dr. Christina Warinner and Dr. Jiawu Xu for their encouragement and valuable knowledge. This includes Dr. Alexandra Obregon-Tito and Raul Tito for their relentless support of my pursuit of a Ph.D., their assistance in sample processing, and their assistance in the Peruvian IRB process and sample collection. I am indebted to both Dr. Krithivas Sankaranarayanan and Allison Mann for their invaluable input regarding my data analysis. Their patience with me during the bioinformatics and statistics steps of this dissertation should be commended. I would like to thank my fellow graduate student cohort for their support and friendship over these last six years. You guys are the best. I need to finally thank my family and friends for their constant encouragement throughout my graduate career and the writing of this dissertation. Thank you specifically to my grandparents Nan and Pete, my sister Emily, and my dad, all of iv whom encouraged me to keep working towards my academic (and life) goals. Lastly, thank you to my mom, who continues to be an unending source of love and support. v Table of Contents Acknowledgements ........................................................................................................ iv List of Tables ................................................................................................................ viii List of Figures ................................................................................................................ ix Abstract .......................................................................................................................... xi Chapter 1 - On Being Human: The Treehouse of Life .................................................1 Our Bacterial Self ...................................................................................................................... 4 Our Viral Self .......................................................................................................................... 19 Other Treehouse Members ...................................................................................................... 29 Final Thoughts on being Human ............................................................................................. 31 Chapter 2 - Viruses in Anthropology...........................................................................34 A Brief History of Viruses (and Microbes) ............................................................................. 35 Viruses as Infectious Agents ................................................................................................... 38 An Anthropological History .................................................................................................... 40 Prions ....................................................................................................................................... 41 Double Stranded DNA Viruses ............................................................................................... 42 Single Stranded DNA Viruses ................................................................................................. 55 Double Stranded RNA Viruses ............................................................................................... 59 Positive Single Stranded RNA Viruses ................................................................................... 61 Negative Single Stranded RNA Viruses .................................................................................. 67 DNA Double Stranded Reverse-Transcribing Viruses ............................................................ 72 RNA Single Stranded Reverse-Transcribing Viruses ............................................................. 72 vi Concluding Statements ............................................................................................................ 80 Chapter 3 - The Geographic Structure of Gut Viruses ..............................................81 Population Background and Sampling Strategies ................................................................... 82 Laboratory Methods ................................................................................................................ 90 Predicted Proteins and Geographic Structure .......................................................................... 94 Amino Acid Sequences and Geographic Structure ................................................................. 99 Conclusions ........................................................................................................................... 102 Chapter 4 - CrAssphage Genomes in the Americas .................................................103 CrAssphage: Lytic or Lysogenic? ......................................................................................... 111 Potential Host Microbiota for crAssphage ............................................................................ 113 CrAssphage Abundance and Host Metabolic Output ............................................................ 120 Final Thoughts on crAssphage .............................................................................................. 123 Chapter 5: Conclusions ...............................................................................................124 References ....................................................................................................................129 Appendix ......................................................................................................................165 vii List of Tables Table 1: Viruses from raw sewage ..................................................................................53 Table 2: Information regarding virus enriched samples ..................................................87 Table 3: Information regarding unenriched virus samples from Oklahoma ...................88 Table 4: Information regarding unenriched virus samples from Peru .............................90 Table 5: CrAssphage abundance from unenriched data ................................................107 Table 6: CrAssphage abundance from enriched data set ...............................................109 Table 7: Contig reads that match crAssphage and other microbes ...............................113 Table 8: CRISPR spacer hits .........................................................................................115 Table 9: CrAssphage abundance and microbial abundance ..........................................117 Table 10: CrAssphage abundance and metabolites .......................................................120 viii List of Figures Figure 1: Almann’s Tree of Life .......................................................................................3 Figure 2: Phylogenetic tree example .................................................................................3 Figure 3: Double stranded DNA viruses .........................................................................42 Figure 4: Eppstein-Barr Virus .........................................................................................46 Figure 5: Single stranded DNA viruses ...........................................................................56 Figure 6: Double stranded RNA viruses .........................................................................60 Figure 7: Positive single stranded RNA viruses ..............................................................62 Figure 8: Negative single stranded RNA viruses ...........................................................67 Figure 9: Double stranded reverse-transcribing DNA viruses ........................................72 Figure 10: Single stranded reverse-transcribing RNA viruses ........................................73 Figure 11: Lentivirus .......................................................................................................75 Figure 12: Map of Peru ....................................................................................................84
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