Integrated Analysis of Bacteroidales and Mitochondrial DNA for Fecal Source Tracking in Environmental Waters

Bibliographic Details
Main Author:	Kapoor, Vikram
Language:	English
Published:	University of Cincinnati / OhioLINK 2014
Subjects:	Environmental Science Fecal source tracking Bacteroidales Mitochondrial DNA Microbial source tracking Fecal indicators Human mitochondrial hypervariable region
Online Access:	http://rave.ohiolink.edu/etdc/view?acc_num=ucin1406821605

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spelling	ndltd-OhioLink-oai-etd.ohiolink.edu-ucin14068216052021-08-03T06:26:29Z Integrated Analysis of Bacteroidales and Mitochondrial DNA for Fecal Source Tracking in Environmental Waters Kapoor, Vikram Environmental Science Fecal source tracking Bacteroidales Mitochondrial DNA Microbial source tracking Fecal indicators Human mitochondrial hypervariable region Identifying the source of surface water fecal contamination is paramount to mitigating pollution and risk to human health. Fecal bacteria such as E. coli have been staple indicator organisms for over a century; however there remains uncertainty with E. coli-based metrics since these bacteria are abundant in the environment. In this study we have shown that human-specific Bacteroidales correlates much more closely to a direct indicator of human waste in the form of human mitochondrial DNA (R =0.62), relative to E. coli (R =0.33), for an urban creek system afflicted by combined sewer overflows. The Duck Creek watershed, which includes Duck Creek, Deerfield Creek and the Little Miami River, was used for sampling due to continued fecal contributions to the Little Miami River recreational waterway. We identified and quantified potential sources of fecal pollution from combined sewer overflows (CSO), and watershed runoff by employing molecular tools for an integrated analysis of Bacteroidales and mitochondrial DNA (mtDNA).The correlation between the presence of human mtDNA, human-specific Bacteroidales and E. coli at sampling sites was studied to design an integrated and reliable fecal source tracking strategy for environmental waters. By comparing a direct indicator (human mtDNA) with bacterial markers, we were able to overcome the limitations of traditional methods which yield false positives from bacteria already living in the stream. Furthermore, we investigated the biodiversity of Bacteroidales 16S rRNA gene sequences derived from some of the water samples to substantiate the evidence of human wastes in the watershed, and to examine their relative abundance among sampling sites. Unlike previous microbial-based correlative investigations, we demonstrate that Bacteroidales closely follow the dynamics of human mitochondrial DNA concentration changes, indicating that these obligate anaerobes are more accurate than E. coli, lending further support to risk overestimation using coliforms.Ion Torrent was used to sequence human mitochondrial hypervariable region II (HVRII) from water samples obtained from CSO sites at different time points accounting for spatial-temporal resolution of human contamination in the watershed. We investigated the occurrence of HVRII allelic frequencies of human mtDNA and used these SNPs to form site-specific genetic barcodes (HVR fingerprint) for evaluating anthropogenic watershed inputs.Altogether, several molecular tools were applied, namely PCR, multiplex PCR, qPCR, and sequencing for elucidating fecal contaminant input and relative contributions. The rigorous sampling procedure followed takes into account wet and dry weather variation, storm event input and diurnal variation. Specifically, the information from this study will not only assist MSDGC with pollution source location and best management practices for mitigating Duck Creek bacterial contributions to the Little Miami River, but will also serve as an invaluable tool for future waste source tracking efforts in general by providing improved methodology. 2014-09-18 English text University of Cincinnati / OhioLINK http://rave.ohiolink.edu/etdc/view?acc_num=ucin1406821605 http://rave.ohiolink.edu/etdc/view?acc_num=ucin1406821605 unrestricted This thesis or dissertation is protected by copyright: all rights reserved. It may not be copied or redistributed beyond the terms of applicable copyright laws.
collection	NDLTD
language	English
sources	NDLTD
topic	Environmental Science Fecal source tracking Bacteroidales Mitochondrial DNA Microbial source tracking Fecal indicators Human mitochondrial hypervariable region
spellingShingle	Environmental Science Fecal source tracking Bacteroidales Mitochondrial DNA Microbial source tracking Fecal indicators Human mitochondrial hypervariable region Kapoor, Vikram Integrated Analysis of Bacteroidales and Mitochondrial DNA for Fecal Source Tracking in Environmental Waters
author	Kapoor, Vikram
author_facet	Kapoor, Vikram
author_sort	Kapoor, Vikram
title	Integrated Analysis of Bacteroidales and Mitochondrial DNA for Fecal Source Tracking in Environmental Waters
title_short	Integrated Analysis of Bacteroidales and Mitochondrial DNA for Fecal Source Tracking in Environmental Waters
title_full	Integrated Analysis of Bacteroidales and Mitochondrial DNA for Fecal Source Tracking in Environmental Waters
title_fullStr	Integrated Analysis of Bacteroidales and Mitochondrial DNA for Fecal Source Tracking in Environmental Waters
title_full_unstemmed	Integrated Analysis of Bacteroidales and Mitochondrial DNA for Fecal Source Tracking in Environmental Waters
title_sort	integrated analysis of bacteroidales and mitochondrial dna for fecal source tracking in environmental waters
publisher	University of Cincinnati / OhioLINK
publishDate	2014
url	http://rave.ohiolink.edu/etdc/view?acc_num=ucin1406821605
work_keys_str_mv	AT kapoorvikram integratedanalysisofbacteroidalesandmitochondrialdnaforfecalsourcetrackinginenvironmentalwaters
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Integrated Analysis of Bacteroidales and Mitochondrial DNA for Fecal Source Tracking in Environmental Waters

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