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A Duplex Digital PCR Assay for Simultaneous Quantification of the Enterococcus spp. and the Human Fecal-associated HF183 Marker in Waters
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Using watershed characteristics to enhance fecal source identification.

John J Hart1, Megan N Jamison2, James N McNair1

  • 1Robert B. Annis Water Resources Institute, 740 West Shoreline Dr, Muskegon, MI, 49441, USA.

Journal of Environmental Management
|March 12, 2023
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Summary

Microbial source tracking (MST) identified human, bovine, and ruminant fecal pollution in waterbodies. Watershed characteristics like soil type and agricultural land use significantly influence contamination risk, aiding in targeted management strategies.

Keywords:
Droplet digital PCR ddPCRFecal indicator bacteriaFecal pollutionMicrobial source trackingWatershed management

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Area of Science:

  • Environmental Science
  • Water Quality Management
  • Microbiology

Background:

  • Fecal pollution is a global issue impacting water safety and ecosystems.
  • Microbial Source Tracking (MST) uses Polymerase Chain Reaction (PCR) to identify fecal pollution origins.
  • Previous MST studies often overlook the influence of watershed characteristics on contamination.

Purpose of the Study:

  • To integrate spatial watershed data with host-associated and general microbial source tracking (MST) markers.
  • To identify the sources of fecal contamination in two distinct watersheds.
  • To investigate the relationship between watershed characteristics and the prevalence of specific fecal markers.

Main Methods:

  • Utilized Polymerase Chain Reaction (PCR) technology, specifically droplet digital PCR (ddPCR), for precise quantification of MST markers.
  • Applied general and host-associated MST markers targeting human (HF183/BacR287), bovine (CowM2), and general ruminant (Rum2Bac) sources.
  • Combined microbial data with spatial watershed characteristics, including soil infiltration rates and land use patterns.

Main Results:

  • All three targeted MST markers (human, bovine, ruminant) were detected across all 25 sampling sites.
  • Bovine and general ruminant markers showed significant associations with specific watershed characteristics.
  • Streams in areas with low-infiltration soils and high agricultural land use exhibited increased fecal contamination risk.

Conclusions:

  • The integration of MST results with watershed characteristics provides a more holistic understanding of fecal contamination.
  • Identifying specific watershed features linked to contamination enables the implementation of more effective best management practices.
  • This approach enhances the ability to mitigate public health risks and protect aquatic environments from fecal pollution.