Microbiology Laboratory

Resuscitation of Old Bacterial Culture

Written by MicroDok

The Resuscitation of Old Bacterial Culture is one of the important technique carried out in the microbiology laboratory. This practice is also obtainable in hospital settings, academic institutes and research facilities where old or stored microbial or bacterial cultures are required for further investigations. Students often ask the question of “when is a bacterial cell considered as dead“? Well, there are various ways to go about it. But the most important thing to know is that: “a bacterium is considered to be dead when it is unable to grow in a medium that normally supports its growth”. Only in this condition can you say an organism or a microbial cell is dead or non-viable. Dormant but viable microbial or bacterial cells are microbes that can grow when the environmental conditions required for its optimal growth are restored. These optimal growth conditions include: nutrients, water, optimal temperature and pressure and availability of growth factors. While aerobic microbes require oxygen to maintain optimal growth; carbon dioxide must be sufficiently provided for the growth of anaerobic microbes.

Before using a stock bacterial culture for further experimental studies, it is very crucial to resuscitate and revive the old bacterial cell which must be in a state of dormancy for a very long period. Resuscitation of the organism is paramount before using the organism, otherwise you might not get the expected result you want from the organism.

To resuscitate the old bacterial cultures, please follow the protocols below:

  1. Inoculate a loopful or speck of the bacterial culture in a 5 ml nutrient broth aseptically.
  2. Incubate the tube in the incubator at 37 degrees celcius for about 6-8 hrs or 18-24 hrs.
  3. After which, transfer a loopful of the content in the test tube onto a nutrient agar plate or any other basal or general purpose medium that will support the growth of the bacteria.
  4. Incubate the nutrient agar plate at 37 degrees celcius overnight.
  5. After incubation, bring out the plates from the incubator.
  6. Observe the culture plate for bacterial growth, usually seen as colonies.
  7. The colonies on the plate will serve as the source of your bacterial cell for your further experimentation.

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