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Related Concept Videos

Microbial Growth Measurement: Indirect Methods01:27

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Estimating microbial growth is essential for understanding population dynamics and environmental adaptations. Indirect methods provide valuable insights by measuring parameters such as turbidity, metabolic activity, and biomass, enabling efficient and reproducible assessments.During exponential growth, microbial cells scatter light proportionally to their biomass, a principle used in turbidity measurements. About one million cells per milliliter produce detectable scattering, which a...
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Direct methods for measuring microbial populations in a culture are essential tools in microbiology, providing quantitative data for various applications. Among these, microscopic counts, plate counts, and serial dilution are widely used techniques, each with unique principles and applications.Microscopic CountsMicroscopic counting involves the use of a Petroff-Hausser chamber, a specialized microscope slide with a grid and defined depth. By observing a liquid culture under a microscope,...
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Analysis of methods for quantifying yeast cell concentration in complex lignocellulosic fermentation processes.

Ruifei Wang1,2, Bettina Lorantfy1,3, Salvatore Fusco1,4

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|May 29, 2021
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Accurate cell quantification in lignocellulose fermentation is difficult. Manual cell counts and colony-forming units (CFU) are reliable methods for measuring yeast cell concentration in these challenging media.

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Area of Science:

  • Biotechnology
  • Industrial Microbiology
  • Process Engineering

Background:

  • Cell mass and viability are critical for fermentation productivity.
  • Lignocellulose-based media present challenges for cell quantification due to particles, auto-fluorescence, color, and turbidity.
  • Existing automated and stain-based methods show limited applicability in these complex media.

Purpose of the Study:

  • To systematically evaluate and validate methods for quantifying total and viable yeast cell concentrations in lignocellulosic media.
  • To optimize parameters for accurate measurements in synthetic lignocellulosic media mimicking industrial conditions.
  • To assess the applicability of validated methods in real-world simultaneous saccharification and fermentation (SSF) processes.

Main Methods:

  • Evaluation of automated cell counting systems and stain-based viability tests.
  • Investigation of manual cell enumeration (hemocytometer), colony-forming units (CFU), quantitative polymerase chain reaction (qPCR), and in situ dielectric spectroscopy.
  • Parameter optimization using synthetic lignocellulosic media.
  • Validation in simultaneous saccharification and fermentation (SSF) experiments using steam-exploded wheat straw.

Main Results:

  • Automated systems and stain-based tests showed limited applicability.
  • Manual hemocytometer counts, CFU enumeration, and dielectric spectroscopy yielded statistically significant calibration models with good predictive capacity.
  • Manual counts and CFU were successfully validated in SSF experiments.
  • Quantitative correlations were found between cell counts, CFU, and in situ permittivity.
  • qPCR demonstrated inconsistent DNA extraction, limiting its reliability.

Conclusions:

  • Manual cell enumeration and CFU plating are robust methods for quantifying yeast in lignocellulosic media.
  • In situ dielectric spectroscopy shows promise for real-time monitoring.
  • qPCR requires further development for reliable DNA extraction from lignocellulosic slurries.
  • Validated cell quantification methods are essential for optimizing and controlling lignocellulose-based fermentation processes.