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Related Concept Videos

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Specialized staining techniques play a vital role in microbiology by enabling the visualization of specific bacterial structures that remain undetectable with standard microscopy methods. These techniques not only enhance the structural visualization of bacterial cells but also provide critical insights into their pathogenicity and classification. Additionally, they support diagnostic and research endeavors in microbiology by identifying key bacterial features.Capsule Staining for Virulence...
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Improving biocide evaluation using propidium monoazide (PMA) viability staining technique.

Xiang Shi1,2, Hasrizal Abd Rahman1,3, Julia R de Rezende4

  • 1Lyell Centre, Heriot-Watt University, Edinburgh, UK.

Scientific Reports
|December 19, 2025
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Summary
This summary is machine-generated.

Propidium monoazide (PMA) viability staining improves biocide efficacy assessment by accurately distinguishing live from dead microbes. This method offers a more reliable evaluation of biocide performance in energy sector applications.

Keywords:
Biocide evaluationPropidium monoazide (PMA)Viability staining

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Area of Science:

  • Microbiology
  • Environmental Science
  • Biotechnology

Background:

  • Chemical biocides are crucial for managing microbial issues in energy sector systems, preventing corrosion and biofouling.
  • Current DNA-based methods for assessing biocide effectiveness struggle to differentiate between live and dead microorganisms, potentially overestimating microbial populations.
  • Propidium monoazide (PMA) viability staining offers a solution to accurately quantify viable microbial cells.

Purpose of the Study:

  • To evaluate the application of PMA viability staining within standard DNA-based workflows for more accurate biocide performance assessment.
  • To investigate the impact of PMA staining on microbial abundance and hydrogen sulfide production in biocide-treated samples.
  • To analyze changes in microbial community structure and diversity following biocide treatment using PMA-enhanced DNA analysis.

Main Methods:

  • A model sulfate-reducing microbial consortium from oilfield produced water was treated with glutaraldehyde (Glut) and tetrakis(hydroxymethyl)phosphonium sulfate (THPS).
  • Propidium monoazide (PMA) staining was performed before DNA extraction, quantitative PCR (qPCR), and 16S rRNA gene amplicon sequencing.
  • Microbial abundance, hydrogen sulfide production, and microbial community properties (alpha and beta diversity, taxonomic composition) were analyzed with and without PMA staining.

Main Results:

  • PMA-treated samples showed microbial abundance at least one order of magnitude lower than untreated samples.
  • Reduced microbial abundance in PMA-treated samples correlated with decreased hydrogen sulfide production, an association not observed without PMA.
  • 16S rRNA gene sequencing revealed significant alterations in microbial diversity and composition in biocide-treated communities only when PMA was applied.

Conclusions:

  • Incorporating PMA viability staining into DNA-based biocide assessment protocols is feasible and significantly enhances accuracy.
  • PMA staining provides a more reliable measure of biocide effectiveness by excluding dead cells, leading to a better understanding of microbial activity.
  • The study recommends adopting PMA staining in future biocide performance studies, particularly for engineered systems in the energy industry, due to its minimal additional effort and substantial benefits.