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Cultivation of Anaerobes

  • Anaerobes are microorganisms that grow in the absence of oxygen.

  • Cultivating anaerobes requires special techniques to create and maintain oxygen-free environments.

  • Two common methods for cultivating anaerobes are the candle jar method and the GasPak jar method.

1) Candle Jar Method

  • Description: A simple and inexpensive method to create a microaerophilic (low oxygen) environment.

Procedure:

  • Place culture plates inside a large, airtight jar.

  • Add a lit candle to the jar and seal the lid.

  • The candle burns, consuming most of the oxygen and increasing the levels of carbon dioxide until the flame extinguishes due to oxygen depletion.

  • Applications: Suitable for growing microaerophilic bacteria and some facultative anaerobes that require reduced oxygen levels but not complete anaerobiosis.

  • Advantages: Easy to set up and inexpensive.

  • Disadvantages: Does not achieve complete anaerobic conditions; only reduces oxygen levels.

2) GasPak Jar Method (Cultivation of Anaerobes)

  • Description: A widely used method for creating an anaerobic environment using chemical reactions.

Procedure:

  • Place culture plates or tubes inside the GasPak jar.

  • Add a GasPak sachet to the jar, which contains chemicals that, when activated, generate hydrogen and carbon dioxide gases.

  • Seal the jar tightly. The hydrogen gas reacts with oxygen in the presence of a palladium catalyst to form water, thus removing oxygen and creating an anaerobic atmosphere.

  • An indicator strip is often used to confirm the absence of oxygen.

  • Applications: Suitable for the cultivation of obligate anaerobes, facultative anaerobes, and microaerophiles.

  • Advantages: Reliable and produces a true anaerobic environment.

  • Disadvantages: Requires the purchase of GasPak sachets and a specialized jar.


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