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Assessment of Fecal Source Pollution in Plum Creek Watershed, Nebraska Using Bacteroidetes-Targeted PCR Assays and Phylogenetic Analysis

Lamendella, Regina
http://rave.ohiolink.edu/etdc/view?acc_num=ucin1141427143

Abstract Details

2006, MS, University of Cincinnati, Engineering : Environmental Engineering.
Recently, 16S rDNA Bacteroidetes-targeted PCR assays were developed to discriminate between ruminant and human fecal pollution. These assays are rapid and relatively inexpensive but have been used in a limited number of environmental applications. In this study, we evaluated the efficacy of human- and ruminant-specific 16S rDNA Bacteroidetes assays in determining the primary sources of fecal pollution in Plum Creek (NE) watershed. The sensitivity and specificity, as well as the temporal and spatial application of these assays were challenged against feces from different animals, water, and sediment samples from this watershed. Phylogenetic analyses of 981 fecal and environmental 16S rDNA clones were also performed to study the diversity of Bacteroidetes in this watershed. On average, the host specific assays indicated that ruminant feces were present in more than one-third of the water samples and in all sampling seasons, with increasing frequency in downstream sites along Plum Creek. The human-targeted assay indicated that only 5% of the water samples were positive for human fecal signals, although a higher percentage of human-associated signals (i.e., 24%) were detected in sediment samples. Phylogenetic analysis of Bacteroidetes 16S rDNA sequences derived from fecal and environmental samples demonstrated the presence of a high level of sequence diversity, with nearly half of the recovered sequences showing less than 97% identity to sequences found in publicly available databases. Approximately, 60% of all clones clustered with yet to be cultured Bacteroidetes species associated with sequences obtained from ruminant feces, further supporting the prevalence of ruminant contamination in this watershed. The preponderance of clones affiliated with uncultured Bacteroidetes strongly suggests that the bacterial diversity of this group is not sufficiently represented by the culturable members for which sequence data are available. The sequencing data also indicated that other sources, including humans, sediments, and wildlife may be significantly contributing to the molecular diversity of fecal Bacteroidetes in this watershed. However, since several clusters contained sequences from multiple sources, future studies must take into consideration the potential cosmopolitan nature of these bacterial populations when assessing fecal pollution sources using Bacteroidetes markers.
Dr. Daniel Oerther (Advisor)
90 p.
Environmental Sciences
Bacteroidetes; Bacteroides-like bacteria; Microbial Source Tracking

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Citations

  • Lamendella, R. (2006). Assessment of Fecal Source Pollution in Plum Creek Watershed, Nebraska Using Bacteroidetes-Targeted PCR Assays and Phylogenetic Analysis [Master's thesis, University of Cincinnati]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1141427143

    APA Style (7th edition)

  • Lamendella, Regina. Assessment of Fecal Source Pollution in Plum Creek Watershed, Nebraska Using Bacteroidetes-Targeted PCR Assays and Phylogenetic Analysis. 2006. University of Cincinnati, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=ucin1141427143.

    MLA Style (8th edition)

  • Lamendella, Regina. "Assessment of Fecal Source Pollution in Plum Creek Watershed, Nebraska Using Bacteroidetes-Targeted PCR Assays and Phylogenetic Analysis." Master's thesis, University of Cincinnati, 2006. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1141427143

    Chicago Manual of Style (17th edition)

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© 2006, all rights reserved.
This open access ETD is published by University of Cincinnati and OhioLINK.