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Studies investigating Escherichia coli strain diversity and demographics in human hosts are frequently inconsistent regarding sampling effort and methodology while current strain typing methods are often expensive or laborious. To rectify these inconsistencies, sampling effort was investigated by comparing the diversity of 15-isolate collections to 100-isolate collections from 3 human subjects. Temporal variation in E. coli strain diversity was also studied by collecting 15 isolates once every 6 months. Additionally, strain identification and diversity collected by different sampling methods (fecal swabs vs. anal swabs collected at different times around defecation) were compared to identify any inherent biases in sampling method. This study employed pyroprinting, a new inexpensive and simple strain typing method using pyrosequencing, to generate DNA fingerprints (or pyroprints) based on the Intergenic Transcribed Spacer sequences in the ribosomal RNA operon to differentiate E. coli strains. Differences in strain diversity were apparent when comparing sampling efforts. The sampling effort investigation suggested that certain subjects hosted very large and highly diverse E. coli strain populations such that even 100 isolates may not fully represent E. coli strain populations in human hosts. Instead, the sampling effort required to accurately represent strain demographics may depend on strain richness and evenness within each host. The temporal investigation yielded similar or greater strain abundance and diversity compared to other typing methods in the literature suggesting pyroprinting is a similarly discriminating tool. When agglomerated over time or by subject, no significant differences in diversity were observed between subjects or between sampling methods despite visible differences in strain richness and evenness. v ACKNOWLEDGMENTS For Arley: I will always love you. The success and completion of this Master's thesis belongs truly to the faithfulness and continued sustenance provided by my Lord and God, Jesus Christ, to whom is owed all my praise. Many thanks to my always encouraging family and supportive friends, but most of all to my husband. Joel, your unconditional love and continued selfless support through this uphill climb gave me strength to endure. Dr. Black and Dr. Kitts, you pushed and challenged me to grow and think in new ways. I am ever grateful for your belief in me. Thank you for all your support and backing.

ESCHERICHIA COLI STRAIN DIVERSITY IN HUMANS: EFFECTS OF SAMPLING EFFORT AND METHODOLOGY