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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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Medium Preparation for the Cultivation of Microorganisms under Strictly Anaerobic/Anoxic Conditions
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Technical note: Evaluation of sampling methods for methane concentration from in vitro fermentation.

Genevieve M D'Souza1, Aaron B Norris2, Luis O Tedeschi1

  • 1Department of Animal Science, Texas A&M University, College Station, TX 77843-2471, USA.

Journal of Animal Science
|August 11, 2022
PubMed
Summary
This summary is machine-generated.

Accurate methane concentration ([CH4]) measurement in in vitro gas production (IVGP) requires accounting for water vapor. Measuring pressure, volume, and temperature and converting to a dry-gas basis ensures more precise results for fermentation studies.

Keywords:
cattleheadspace gasin vitro gas productionmethaneruminant

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Area of Science:

  • Agricultural Science
  • Animal Science
  • Environmental Science

Background:

  • In vitro gas production (IVGP) is a common method for evaluating feed fermentation in ruminants.
  • Accurate measurement of methane concentration ([CH4]) in IVGP headspace is crucial for understanding fermentation dynamics and greenhouse gas emissions.
  • Existing protocols for sampling and analyzing IVGP headspace gas may influence methane concentration measurements.

Purpose of the Study:

  • To evaluate different methods for sampling methane concentration ([CH4]) from in vitro gas production (IVGP) headspace.
  • To determine if direct headspace sampling or exetainer sampling yields comparable [CH4] results.
  • To assess the impact of water vapor on [CH4] measurements and compare results on a dry-gas basis.

Main Methods:

  • Compared headspace gas sampling methods: direct measurement (DIRECT), cooling on ice (ICE), and refrigeration (FRIDGE).
  • Measured pressure (P), volume (V), and temperature (T) of IVGP headspace gas.
  • Quantified methane concentration ([CH4]) using gas chromatography and calculated dry-gas basis values accounting for water vapor pressure (Pw).

Main Results:

  • No significant difference was found between exetainer (EXET) and bottle (BOTT) headspace [CH4] sampling (P = 0.28).
  • Relative humidity (RH) of IVGP slurry was 100%, confirming gas saturation with water vapor.
  • Significant differences in P, V, T, and [CH4] were observed among DIRECT, ICE, and FRIDGE methods (P < 0.01).

Conclusions:

  • Current methods for assessing IVGP methane concentration show variability, necessitating careful consideration.
  • Direct measurement of P, V, and T during headspace sampling is recommended for accurate [CH4] determination.
  • Converting [CH4] to a dry-gas basis using measured parameters is essential for removing volume inflation caused by water vapor.