Life Without Oxygen: The Anaerobic Chamber

Is life possible without oxygen?

While many science fiction films have popularized the idea that oxygen is an essential prerequisite for life in general, even on our planet there are organisms that do not require oxygen or for which it is even lethal!

The characteristic of these organisms is called “anaerobiosis” (from the Greek aero and bios, therefore “life without air”) and the anaerobic chamber is the instrument that modern scientists use to study them.

What is the Anaerobic Chamber?

An anaerobic chamber is an isolated environment designed to drastically reduce or even completely eliminate oxygen levels within it. In addition to oxygen, other environmental parameters such as temperature and relative humidity can usually be monitored and adjusted within the anaerobic chamber .

How an Anaerobic Chamber is Made

There are different models of anaerobic chamber but some features are common to all.

First, the interior of the chamber must be hermetically sealed from the outside for obvious reasons. Therefore, to handle the samples introduced inside, anaerobic chambers often have gloved doors that allow operators to intervene on the bacterial cultures.

Other essential elements of the chamber are the control of the internal atmosphere and oxygen.

The oxygen-free environment is in fact obtained by filling the space with some inert gases, usually nitrogen or argon.

The anaerobic chambers are also equipped with monitoring systems that constantly monitor both the presence of oxygen inside themand other environmental parameters such as temperature and relative humidity, in order to guarantee the precision and reliability of the test.

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What are Anaerobic Chambers Used For?

Anaerobic chambers find application in various scientific fields but above all in medical and pharmacological because they constitute the ideal environment for cultivating and studying certain species of anaerobic bacteria, organisms that thrive in the absence of oxygen.

A more in-depth analysis of this fascinating type of organism can be found here. 

In the chemical industry, however, they are used for the handling of oxygen-sensitive chemicals and in the production of materials that require an oxygen-free environment to maintain their integrity.

Non-Anaerobic Growth Chambers

Despite their fundamental importance in the study of anaerobic organisms, anaerobic chambers are only one type of a larger class of laboratory instruments called growth chambers.

Growth chambers are defined as all laboratory instruments suitable for culturing microorganisms and plants. All growth chambers implement systems for regulating environmental parameters, including temperature and relative humidity. The anaerobic chamber, as we have seen, also has oxygen regulation systems, while more traditional growth chambers feature technologically advanced lighting systems that simulate sunlight.

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The FDM Growth Chamber

For over 70 years, FDM – Environmental Makers has been designing laboratory instruments capable of simulating the most varied environmental conditions in the field of climate testing.

Our growth chambers reproduce the ideal environment for growing aerobic organisms such as bacterial and plant cultures. We also build custom growth chambers, as we did for the University of Turin. You can read that story here.

For any further questions or concerns, please do not hesitate to contact us.

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Reference Standards

• ISO 14853 — Plastics: Determination of the ultimate anaerobic biodegradation of plastic materials in an aqueous system, aqueous phase method.
• DIN EN ISO 11734 — Water quality: Evaluation of the ultimate anaerobic biodegradability of organic compounds in digested sludge, test on digested sludge.
• ASTM D5210 — Standard Test Method for Determining the Anaerobic Biodegradation of Plastic Materials in the Presence of Municipal Sewage Sludge.
• ISO 15985 — Plastics: Determination of the ultimate anaerobic biodegradation under high-solids anaerobic-digestion conditions.
• CLSI M11 — Methods for Antimicrobial Susceptibility Testing of Anaerobic Bacteria, clinical microbiology reference.

Authoritative Sources

• ISO – International Organization for Standardization. Maintains ISO 14853 and ISO 15985 on anaerobic biodegradability.
• ASTM D5210. American Standard for Anaerobic Biodegradability of Plastics in Sewage Sludge.
• CLSI – Clinical and Laboratory Standards Institute. CLSI M11 for antimicrobial susceptibility of anaerobic bacteria has been published.

Learn more: discover our controlled atmosphere CO2 testing chamber, designed and manufactured by FDM Environment Makers.

Frequently Asked Questions about the Anaerobic Chamber

What methods are there to create anaerobiosis?

Three main methods with increasing levels of rigor. Gaspak systems (Becton Dickinson, Oxoid AnaeroGen): gas-generating bags in sealed jars, consume residual O2 via ascorbic acid/Na-borohydride reaction producing final atmosphere. Hungate technique: glass tubes pressurized with pure anaerobic gas (N2, N2/CO2, H2/CO2), closed with butyl stoppers, access with syringes and purged hypodermic needles. Achieves levels of anaerobic Glovebox (Coy Laboratory, Whitley A35, Don Whitley): rigid enclosure with pressure seal, atmosphere maintained with 3-5% H2 + N2 or N2/CO2, Pd catalyst that consumes residual O2 H2 -> H2O. Levels

Typical anaerobic gas composition?

The gas mixture depends on the application. Anaerobic clinical workstation (CLSI M11): 85% N2 + 10% CO2 + 5% H2, where H2 is used for catalytic O2 consumption via Pd and CO2 for buffering Brucella media or Brain Heart Infusion. Methanogens/archaea basic research: 80% N2 + 20% CO2 with bulk addition of H2 as a metabolic substrate. Biodegradation ISO 14853: 70% N2 + 30% CO2 to maintain stable inoculum pH via bicarbonate equilibrium. Clostridium culture : pre-purge pure N2 for 30 min, then working atmosphere of 90% N2 + 10% CO2 with H2S present as a natural bacterial metabolite. The gas quality is high: N2 5.0 (99.999%), CO2 food grade 4.5, H2 ultra pure 5.0 with residual O2 < 1 ppm, all gases pass through oxisorb filters for further deoxygenation. Atmospheric monitoring with electrochemical O2 sensors (limit 1 ppm) and CO2 NDIR (+/- 0.5%) must be continuous with an alarm in case of exceedance.

What is the redox potential?

The redox potential Eh measures the oxidation-reduction state of the aqueous environment, expressed in mV with respect to a standard hydrogen electrode (SHE). Water-saturated air has Eh +400/+500 mV, strictly anaerobic environment Eh -400 mV (bovine rumen, digested sludge). For the cultivation of strict anaerobes (Clostridium botulinum, methanogens, Desulfovibrio) Eh reducing agents: cysteine-HCl 0.05%, Na2S 0.025%, thioglycollate 0.075%, titanium(III) citrate 1-4 mM for extreme conditions. A redox indicator such as resazurin (pink -> colorless at Eh -110 mV) visually indicates the anaerobic nature of the medium. The Eh measurement must take into account temperature (+1 C reduces Eh by ~1 mV) and pH (Nernst correlation). ISO 14853 biodegradation software calculates the Biochemical Methane Potential BMP by integrating CH4 + CO2 production in gas headspace against reference pressure and temperature, with deduction of endogenous control (inoculum without substrate).

Typical applications: Clostridium, methanogens, bacteroides?

Clinical applications of the anaerobic chamber include: diagnosis of C. difficile in post-antibiotic diarrhea (culture on CCFA agar, PCR for toxins A/B), isolation of Bacteroides fragilis from abdominal abscesses, identification of Fusobacterium nucleatum in ENT infections, and culture of Peptostreptococcus in endometritis. Incubation time 48-72 h at 37°C with MALDI-TOF colony reading for rapid identification. Biotechnological applications include: study of methanogens Methanobacterium formicicum and Methanosarcina for biogas from anaerobic digesters, butanol production with C. acetobutylicum ABE fermentation, succinic acid production with Actinobacillus succinogenes , and biosynthesis of B12 cofactors with Propionibacterium . Environmental applications : Biodegradation of compostable plastics (PLA, PHA, starch-based) with inoculation from mesophilic digestate at 35°C / thermophilic digestate at 55°C for 60–90 days, quantification of CO2 + CH4 vs. reference cellulose, calculation of % mineralization. The anaerobic research chamber must have a volume of 150–500 L, two independent airlocks, an H2S scrubbing system for operator protection, and continuous data logging of atmosphere, temperature, and humidity.