The use of PCR and T-RFLP as a means of identifying sources of fecal bacteria pollution in the tidal creeks of New Hanover County, North Carolina

UNCW Author/Contributor (non-UNCW co-authors, if there are any, appear on document)
Mary I. H. Spivey (Creator)
The University of North Carolina Wilmington (UNCW )
Web Site:
Michael Mallin

Abstract: Tidal creeks are deteriorating more each day as a consequence of increased pollution caused by a boom in urbanization, which has intensified the anthropogenic impact on these ecosystems. These systems are subject to anthropogenic pollutants such as nutrients, pesticides, and fecal bacteria, which can cause loss of habitat, anoxia, and shellfish bed closures. In 2001, 13,410 recreational area closings and advisories were issued nationwide. Nearly 90% of these were due to the level of fecal coliform bacteria in the water. The standard method for fecal coliform pollution measurement enumerates, but does not distinguish between sources. Using standard methods for fecal coliform bacteria enumeration, and molecular methods of PCR and T-RFLP for bacterial source tracking using the genera Bacteroides as a target, it was possible to identify areas with high level of fecal coliform bacteria pollution as well as distinguish between human, canine and ruminant sources. Of the 54 samples, less than 23% were positive for canine fecal bacteria DNA by PCR, indicating fairly successful domestic animal waste management. Ruminant sources were found in 12 of the 54 PCR samples, mainly in the upstream sampling areas and near a known horse farm. Human fecal bacteria DNA was found in 18% of the PCR samples. T-RFLP produced 40 peaks, each corresponding to a bacterial taxon. Using the MiCA T-RFLP PAT+, it was possible to identify 13 of the 40 peaks, 11 of which were human-specific The presence of human-specific DNA is of particular concern. New Hanover County has been plagued with sewer-system failures. The presence of human-borne Bacteroides DNA is indicative of either continued sewer-line problems, septic system failures, or a general persistence in the bacteria itself.

Additional Information

A Thesis Submitted to the University of North Carolina Wilmington in Partial Fulfillment of the Requirements for the Degree of Master of Science
Language: English
Date: 2009
Bacterial pollution of water--North Carolina--New Hanover County, Feces--Microbiology--North Carolina--New Hanover County, Polymerase chain reaction--Diagnostic use, Water--Pollution--North Carolina--New Hanover County
Water -- Pollution -- North Carolina -- New Hanover County
Feces -- Microbiology -- North Carolina -- New Hanover County
Bacterial pollution of water -- North Carolina -- New Hanover County
Polymerase chain reaction -- Diagnostic use

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