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Assessing accuracy and specificity of faecal source library for microbial source-tracking, using SourceTracker as

Timothy J Y Lim1, Yussi M Palacios Delgado1, Anna Lintern1

  • 1Department of Civil & Environmental Engineering, Monash University, Clayton, VIC 3800, Australia.

Bioinformatics Advances
|May 21, 2025
PubMed
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Assessing faecal source library quality is crucial for microbial source tracking (MST). Our method validates library quality, improving accuracy for environmental monitoring and public health.

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Area of Science:

  • Environmental microbiology
  • Bioinformatics
  • Molecular ecology

Background:

  • Microbial source tracking (MST) relies on accurate faecal source libraries.
  • Library quality is often overlooked, impacting MST reliability.

Purpose of the Study:

  • To develop and demonstrate an assessment approach for validating amplicon-derived faecal source libraries.
  • To evaluate the impact of library quality and sample size on MST accuracy.

Main Methods:

  • Leave-one-out (LOO) analysis to assess source category assignment accuracy.
  • Quality control procedures for sample retention/removal/grouping.
  • Sample size assessment to determine sufficient characterization of source fingerprints.

Main Results:

  • LOO analysis revealed 15.5% of samples were incorrectly assigned, with high error rates for birds and wallabies.
  • Increasing sample size improved source identification accuracy, but accuracy plateaued.
  • The study highlights the importance of library quality assessment for reliable MST.

Conclusions:

  • A robust library assessment method is essential for library-dependent MST.
  • Understanding library limitations is critical before applying MST for environmental assessments.
  • This approach enhances the reliability of microbial source tracking studies.