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HASELHORST-THESIS.Pdf (2.779Mb) TYPICAL COLLEGE STUDENT DIET FOOD DERIVED MICROORGANISMS AND THEIR RELATION TO THE HUMAN GASTROINTESTINAL MICROFLORA A Senior Scholar Thesis by ALEXANDRIA JOANN HASELHORST Submitted to the Office of Undergraduate Research Texas A&M University in partial fulfillment of the requirements for the designation as UNDERGRADUATE RESEARCH SCHOLAR April 2010 Major: Nutritional Sciences TYPICAL COLLEGE STUDENT DIET FOOD DERIVED MICROORGANISMS AND THEIR RELATION TO THE HUMAN GASTROINTESTINAL MICROFLORA A Senior Scholar Thesis by ALEXANDRIA JOANN HASELHORST Submitted to the Office of Undergraduate Research Texas A&M University in partial fulfillment of the requirements for the designation as UNDERGRADUATE RESEARCH SCHOLAR Approved by: Research Advisor: Dr. Suresh Pillai Associate Dean for Undergraduate Research: Robert C. Webb April 2010 Major: Nutritional Sciences iii ABSTRACT Typical College Student Diet Food Derived Microorganisms and Their Relation to the Human Gastrointestinal Microflora. (April 2010) Alexandria Joann Haselhorst Department of Nutrition Texas A&M University Research Advisor: Dr. Suresh Pillai Department of Poultry Science & Nutrition and Food Science Current research in the field of obesity has shown that obese people generally have a higher percentage of Firmicutes compared to Bacteroidetes in their gastrointestinal tracts than lean people. The goal of this study was to assess whether or not different foods contain unique bacterial profiles and if there is a correlation between foods and the types of bacteria that may be present. This study was based on diets that are typically consumed by college students. Six meals, two breakfasts, two lunches, and two dinners were created based off a class project that detailed the diet of Texas A&M University students. These meals consisted of a banana, cereal, and orange juice; a bagel, breakfast burrito and coffee; a turkey sub sandwich, chips and a soda; macaroni and cheese, carrots and water; pepperoni pizza, salad and water; and finally a cheeseburger, fries and a soda. All the foods were purchased either on campus dining halls, or stores and fast food restaurants in College Station, Texas. Portions of the food samples were plated on Tryptic Soy Agar media and Brucella Blood Agar to determine the aerobic and iv anaerobic bacterial loads respectively. The total microbioial communities were extracted from defined amounts of the different food samples and the 16s rRNA genetic diversity were analyzed pyrosequencing. It was found that each food does have a unique bacterial profile. There was, however, no correlation between the nutritional content of the foods and presence of specific bacterial groups. It was determined that on an average, a college student will consume via foods a minimum of about 6.38x1010 bacterial sequences of bacteria each year, and about 2.55x1011 sequences of bacteria during an average 4-year college career. The data implies that foods contribute not only nutrients to the human body but can also be a major source for the introduction of microbial populations into the gastrointestinal tract. Since alterations in the gastrointestinal populations do alter host-microbiota interactions which in turn affect metabolic syndromes, this study illustrates one pathway through which microbial populations are adding to the diversity within the gastrointestinal system. v DEDICATION I would like to dedicate this thesis to my parents who have always believed I could accomplish anything I set my mind to and to my brother Erik for encouraging me to do so. vi ACKNOWLEDGMENTS I would like to thank Dr. Suresh Pillai most of all for all his time and encouragement over this past two years. Also, I would like to thank Dr. Palmy Jesudhasan, Katherine Grace McElhany and the rest of the lab for all their help and resources. I would like to thank Dr. Scot Dowd for performing pyrosequencing for this project. Finally, I would like to thank the Office of Undergraduate Research for providing students the opportunity of undergraduate research and allowing me to take part in the program. vii NOMENCLATURE BBA Brucella Blood Agar CFU Colony Forming Unit PCR Polymerase Chain Reaction TSA Tryptic Soy Agar viii TABLE OF CONTENTS Page ABSTRACT ....................................................................................................................... iii DEDICATION .................................................................................................................... v ACKNOWLEDGMENTS .................................................................................................. vi NOMENCLATURE .......................................................................................................... vii TABLE OF CONTENTS ................................................................................................. viii LIST OF FIGURES ............................................................................................................. x LIST OF TABLES ............................................................................................................ xii CHAPTER I INTRODUCTION ....................................................................................... 1 II METHODS .................................................................................................. 7 Food samples and sample processing .............................................. 7 Aerobic and anaerobic plate counts ................................................ 8 Molecular methods .......................................................................... 9 III RESULTS .................................................................................................. 11 Culture-based methods .................................................................. 11 Pyrosequencing results .................................................................. 13 Discussion ..................................................................................... 60 IV SUMMARY .............................................................................................. 67 REFERENCES .................................................................................................................. 68 APPENDIX A ................................................................................................................... 70 APPENDIX B ................................................................................................................... 71 APPENDIX C ................................................................................................................... 73 ix APPENDIX D ................................................................................................................... 80 CONTACT INFORMATION ........................................................................................... 82 x LIST OF FIGURES FIGURE Page 1 Phylogenetic analysis of the metagenomic content of breakfast one determined by pyrosequencing ...................................................... 15 2 Phylogenetic analysis of the metagenomic content of cereal determined by pyrosequencing .................................................................. 17 3 Phylogenetic analysis of the metagenomic content of banana determined by pyrosequencing ................................................................ 19 4 Phylogenetic analysis of the metagenomic content of water determined by pyrosequencing ................................................................... 22 5 Phylogenetic analysis of the metagenomic content of breakfast two determined by pyrosequencing ...................................................... 23 6 Phylogenetic analysis of the metagenomic content of breakfast burrito determined by pyrosequencing ................................................. 26 7 Phylogenetic analysis of the metagenomic content of bagel determined by pyrosequencing ................................................................... 28 8 Phylogenetic analysis of the metagenomic content of coffee determined by pyrosequencing .................................................................. 31 9 Phylogenetic analysis of the metagenomic content of lunch one determined by pyrosequencing ............................................................ 33 10 Phylogenetic analysis of the metagenomic content of macaroni & cheese determined by pyrosequencing ............................................. 35 11 Phylogenetic analysis of the metagenomic content of apple determined by pyrosequencing ................................................................... 38 12 Phylogenetic analysis of the metagenomic content of lunch two determined by pyrosequencing ............................................................ 39 13 Phylogenetic analysis of the metagenomic content of turkey sandwich determined by pyrosequencing ................................................. 42 xi FIGURE Page 14 Phylogenetic analysis of the metagenomic content of carrots by pyrosequencing .................................................................................... 44 15 Phylogenetic analysis of the metagenomic content of soda determined by pyrosequencing .................................................................... 46 16 Phylogenetic analysis of the metagenomic content of dinner one determined by pyrosequencing ........................................................... 48 17 Phylogenetic analysis of the metagenomic content of pepperoni pizza determined by pyrosequencing .................................................
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