Semester

Spring

Date of Graduation

2010

Document Type

Thesis

Degree Type

MS

College

Statler College of Engineering and Mineral Resources

Department

Civil and Environmental Engineering

Committee Chair

Benoit Van Aken.

Abstract

Microbial community structure in groundwater is extremely complex and constitutes a fingerprint of water. Terminal restriction fragment length polymorphism (T-RFLP) analysis is a polymerase chain reaction (PCR)-based fingerprinting method that is commonly used for comparative microbial community analysis, and to track specific bacterial markers in complex environments.;The Objective of this research was to propose an innovative approach for tracking the origin of groundwater, microbial source tracking and origin of surface water, based on the specific structure of the microbial community using T-RFLP analysis, which can be used for microbial source tracking.;The first specific aim was to validate the use of T-RFLP profiles of deep and shallow waters in Huntingdon, PA, Great Valley, WV and Berkeley springs, WV, as a marker of the groundwater source. Alternatively, a 16S ribosomal DNA genomic library was constructed to identify bacterial species that can inform about the source of groundwater. It was observed that clustering based on screening a clone library gave more reliable information of the groundwater source than the T-RFLP profiles.;The second specific aim focused on the characterization of the microbial community from different streams along the Left Fork Mud River watershed in Lincoln County, WV. T-RFLP analyses were carried out for determining the source of contaminated water. Results showed that different streams of the Mud River watershed has a specific microbial terminal restriction fragment (TRF) profiles, which helps in determining the origin of water.;The third specific aim was to characterize the entire microbial community in several streams in Corridor-H watersheds, WV using T-RFLP analysis. It was observed that water samples collected from different locations of the same stream carried a very specific signature that can be used for microbial source tracking analysis. The T-RFLP method application on spring waters and Mud River watersheds was based on the preliminary results obtained from the analysis of the microbial community structure of several streams in Corridor-H watersheds.;Our results, thus provide the evidence that natural waters has a 'microbial signature' (T-RFLP profile) that can be used as a marker for pathogens and groundwater source tracking.

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