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Colorimetric Paper-based Detection of Escherichia coli, Salmonella spp., and Listeria monocytogenes from Large Volumes of Agricultural Water
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Comparative Assessment of Wastewater-Based Surveillance Normalization Methods to Improve Pathogen Monitoring in Rural

Amanda Darling1,2, Benjamin C Davis1,3, Thomas Byrne4

  • 1Department of Population Health Sciences, Virginia Tech, Blacksburg, Virginia 24061, United States.

Environmental Science & Technology
|May 27, 2025
PubMed
Summary
This summary is machine-generated.

Wastewater surveillance normalization methods are affected by inflow and infiltration (I&I). Human fecal markers offer a robust normalization approach for sewersheds with significant I&I, improving data reliability.

Keywords:
inflow and infiltrationnormalizationpublic healthrural healthwastewater-based surveillance

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Area of Science:

  • Environmental microbiology
  • Public health surveillance
  • Wastewater engineering

Background:

  • In-sewer dynamics significantly impact wastewater surveillance accuracy.
  • Conventional normalization methods may fail in sewersheds with substantial inflow and infiltration (I&I).

Purpose of the Study:

  • To evaluate the effectiveness of various normalization approaches for wastewater pathogen signals.
  • To assess the impact of I&I on normalization strategies in a rural sewershed.

Main Methods:

  • Monthly wastewater sampling from 2022-2023 at a WWTP influent and 11 sewer nodes.
  • Quantification of SARS-CoV-2, norovirus GII, and rotavirus.
  • Comparison of normalization using flow, population, physicochemical parameters, and human fecal markers (crAssphage, HF183, mtDNA).

Main Results:

  • Physicochemical normalization introduced variability and correlated with precipitation in I&I-impacted systems.
  • Viral signals normalized with physicochemical parameters showed less association with actual pathogen levels.
  • Normalization using human fecal markers demonstrated greater robustness in the presence of I&I.

Conclusions:

  • Standard normalization methods using physicochemical parameters are unreliable in sewersheds with high I&I.
  • Human fecal markers provide a more stable and accurate normalization strategy for wastewater-based epidemiology in I&I-affected areas.
  • Accurate normalization is crucial for reliable wastewater surveillance, especially in diverse sewer system conditions.