Document Type

Thesis

Date of Award

12-2005

School/College

College of Science, Engineering, and Technology (COSET)

Degree Name

MS in Biology

First Advisor

Professor Olufisayo Jejelowo

Abstract

Microorganisms have been reported in closed environments at different NASA facilities, from past NASA missions, and in the Russian Space Station MIR. Microorganisms were the first inhabitants of the International Space Station (ISS). They can impact human health and plant health, and cause damage to construction materials in space as well as on Earth. Information on microorganisms and their impact on environmental safety during Human Exploration Missions is important, especially for missions of longer duration. The main objective of this project was to develop an easy to use, rapid, sensitive and accurate model for the identification of relevant fungal species using molecular techniques such as polymerase chain reaction, Sanger sequencing and a novel technology, Pyrosequencing. Conventional methods of identification, such as culturing and morphological analysis, have been proven problematic. These methods are typically time-consuming and laborious. Additionally, microscopic analysis and identification based on morphology requires a highly skilled technician in order to avoid misidentification. Preliminary experiments based on initial isolates received indicate some discrepancy in identification. Nucleic acid based identification provides great accuracy and efficiency. Pyrosequencing technology and a multiple sequencing primer method designed for the detection of multiple species in a sample were therefore used for identifying the relevant fungal species. 2

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