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Related Experiment Video

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The Cultivation, Growth, and Viability of Lactic Acid Bacteria: A Quality Control Perspective
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Collaborative cytometric inter-laboratory ring test for probiotics quantification.

Peter Lüttge Jordal1, Marcos González Diaz1, Carlotta Morazzoni2

  • 1SBT Instruments A/S, Herlev, Denmark.

Frontiers in Microbiology
|November 29, 2023
PubMed
Summary
This summary is machine-generated.

Accurate enumeration of probiotics is crucial for health benefits. This study shows flow cytometry methods, including fluorescence and impedance flow cytometry, reliably quantify live bacterial cells, offering a culture-independent alternative.

Keywords:
culture-independentdormantelectrical impedance spectroscopy flow cytometry (EIS-FC)fluorescence flow cytometryimpedance flow cytometryprobioticsviable but non-culturable

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Area of Science:

  • Microbiology
  • Analytical Chemistry
  • Biotechnology

Background:

  • Probiotics, live microorganisms conferring health benefits, require accurate enumeration for efficacy.
  • Current gold standard, colony forming units (CFU), relies on cultivability, potentially underestimating viable cells.
  • The Viable But Non-Culturable (VBNC) state highlights limitations of culture-dependent viability assessments.

Purpose of the Study:

  • To evaluate flow cytometry methods for reliable bacterial enumeration in probiotic products.
  • To compare fluorescence flow cytometry (FFC) and impedance flow cytometry (IFC) for quantifying probiotic bacteria.
  • To assess the robustness and agreement of these culture-independent methods.

Main Methods:

  • An inter-laboratory ring test was conducted for cytometric bacterial quantification.
  • Membrane integrity was assessed using fluorescence flow cytometry (FFC) and impedance flow cytometry (IFC).
  • Six distinct probiotic bacterial species (3 Lactobacillus, 3 Bifidobacterium) were tested.

Main Results:

  • The FFC method demonstrated robustness across different equipment, procedures, materials, and operators.
  • After optimization, the IFC method showed good agreement with FFC results for total and live cell counts.
  • Both FFC and IFC provided consistent results for the tested probiotic strains.

Conclusions:

  • Flow cytometry offers a reliable, precise, and user-friendly culture-independent method for bacterial enumeration.
  • FFC and IFC are suitable alternatives to traditional culture-based methods for assessing probiotic viability.
  • These findings support the use of flow cytometry for accurate quality control of probiotic products.