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Toolbox approaches using molecular markers and 16S rDNA amplicon datasets for the identification of fecal pollution in surface water
Journal article   Open access   Peer reviewed

Toolbox approaches using molecular markers and 16S rDNA amplicon datasets for the identification of fecal pollution in surface water

Warish Ahmed, C Staley, M J Sadowsky, P Gyawali, Jatinder P S Sidhu, A Palmer, D J Beale and S Toze
Applied and Environmental Microbiology, Vol.81(20), pp.7067-7077
2015
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https://doi.org/10.1128/AEM.02032-15View
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Abstract

microbial source tracking fecal indicator bacteria operational taxonomic units (OTU) molecular markers wastewater real-time PCR
In this study, host-associated molecular markers and bacterial 16S rDNA community analysis using high throughput sequencing were used to identify the sources of fecal pollution in environmental waters in Brisbane, Australia. A total of 92 fecal and composite wastewater samples were collected from different host groups (cat, cattle, dog, horse, human and kangaroo). In all, 18 water samples were collected from six sites (BR1-BR6) along the Brisbane River, Qld, Australia. Bacterial communities in fecal/wastewater and river water samples were sequenced. Water samples were also tested for the presence of avian- (GFD), cattle- (CowM3), horse-, and human- (HF183) associated molecular markers to provide multiple lines of evidence on the possible presence of fecal pollution associated with specific host(s). Among the 18 water samples tested, 83%, 33%, 17%, and 17% were real-time PCR positive for the GFD, HF183, CowM3, and horse markers, respectively. The potential sources of fecal pollution in water samples from the studied river using DNA sequencing tended to show relatively small contributions from wastewater treatment plants (WWTPs) (up to 13% of sequence reads). Contributions from other animal sources were rarely detected and were very small (< 3% of sequence reads). Source contributions determined via sequence analysis versus molecular markers showed variable agreement. Lack of relationships was also observed among fecal indicator bacteria (FIB), host-associated molecular markers, and 16S rDNA community analysis. Nonetheless, we show that bacterial community and host-associated molecular marker analysis can be combined to identify potential sources of fecal pollution in an urban river. This study is a proof of concept and based on the results, we recommend using bacterial community analysis (where possible) along with the polymerase chain reaction (PCR) detection or quantification of host-associated molecular marker to provide information on the source(s) of fecal pollution in waterways.

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Domestic collaboration
International collaboration
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Biotechnology & Applied Microbiology
Microbiology

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#3 Good Health and Well-Being
#6 Clean Water and Sanitation
#11 Sustainable Cities and Communities
#14 Life Below Water

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