Compositional and temporal stability of fecal taxon libraries for use with SourceTracker in sub-tropical catchments

Christian O'Dea, Qian Zhang, Christopher Staley, Nicole Masters, Anna Kuballa, Paul Fisher, Cameron Veal, H. Stratton, Michael J. Sadowsky, Warish Ahmed, Mohammad Katouli

Research output: Contribution to journalArticlepeer-review

12 Scopus citations


Characterization of microbial communities using high-throughput amplicon sequencing is an emerging approach for microbial source tracking of fecal pollution. This study used SourceTracker software to examine temporal and geographical variability of fecal bacterial community profiles to identify pollutant sources in three freshwater catchments in sub-tropical Australia. Fecal bacterial communities from 10 animal species, humans, and composite wastewater samples from six sewage treatment plants were characterized and compared to freshwater samples using Illumina amplicon sequencing of the V5–V6 regions of the 16S rRNA gene. Source contributions were calculated in SourceTracker using new fecal taxon libraries as well as previously generated libraries to determine the effects of geographic and temporal variability on source assignments. SourceTracker determined 16S rRNA bacterial communites within freshwater samples, shared taxonomic similarities to that of wastewater at low levels (typically <3%). SourceTraker also predicted occasional fecal detection of deer and flying fox sources in the water samples. No significant differences in source contributions were observed within sequences from current and previously characterized fecal samples (P ≥ 0.107). However, significant differences were observed between previously characterized and newly characterized source communities (ANOSIM P ≤ 0.001), which shared <15% community composition. Results suggest temporal instability of fecal taxon libraries among tested sources and highlight continual evaluation of community-based MST using confirmatory qPCR analyses of marker genes.

Original languageEnglish (US)
Article number114967
JournalWater Research
StatePublished - Nov 15 2019

Bibliographical note

Funding Information:
We want to acknowledge Water Research Australia and notably Carolyn Bellamy for enabling the collaboration of this project with Seqwater. We would also like to thank and acknowledge Dr Miranda Emily Vidgen of the Queensland Institute of Medical Research (QIMR) for her advice on animal DNA extraction methods, Robert Smith of the University of the Sunshine Coast (USC) for assisting in collection of animal fecal samples, Rebecca Veca of the University of the Sunshine Coast for collaborations in human sample acquisition, and Makeely Blandford of the University of the Sunshine Coast for assisting in field sampling, and their extractions.

Publisher Copyright:
© 2019


  • 16S rRNA
  • Amplicon sequencing
  • Microbial source tracking
  • SourceTracker
  • Wastewater


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