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Cell Biology JoVE (Open Access) Citable · DOI

Medium Preparation for the Cultivation of Microorganisms under Strictly Anaerobic/Anoxic Conditions

DOI: 10.3791/60155-v
What you'll learn
  • ✓ Prepare anaerobic medium using reducing agents
  • ✓ Fill and seal cultivation flasks oxygen-free
  • ✓ Inoculate medium with anaerobic mixed culture
  • ✓ Quantify gas and volatile fatty acids
Protocol

Biopharma Insights As obligate anaerobic organisms are unable to grow upon oxygen exposure, the use of anaerobic culturing techniques is indispensable. Here, we demonstrate a simple and effective method to cultivate a mixed culture derived from a biogas plant from media preparation to gas and volatile fatty acid quantification.

Difficulty
intermediate
Total time
~2-3 days
Model organism
Mixed anaerobic microbial culture (biogas plant inoculum)
Biosafety
BSL-1

Steps

1
Prepare medium and fill cultivation flasks

Prepare the anaerobic medium with reducing agents and dispense it into cultivation flasks while maintaining anoxic conditions.

▶ 00:39
2
Inoculate the anaerobic medium

Aseptically transfer the biogas plant-derived mixed culture into the prepared anaerobic medium.

▶ 02:29
3
Incubate, sample, and analyze

Incubate the cultures under anaerobic conditions, collect samples over time, and quantify gas and volatile fatty acids.

▶ 03:38
4
Review representative anaerobic cultivation results

Examine example results showing successful growth and metabolic activity of the anaerobic mixed culture.

▶ 04:57
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