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Cultivation and storage

Storage

Luminescent bacteria can be stored as standard glycerol stab cultures.
This is the method of preparing glycerol stab cultures of bacteria:

Cultivation of Luminescent Bacteria

Cultivation of luminous bacteria is processed on the special medium "BOSS"
This is the method of preparation the medium "BOSS":

  30 g NaCl
  1 g Glycerol
  10 g Peptone (Bacto-Peptone)
  3 g Beef Extract
  (to obtain solid medium you should add 15 g of Bacto-Agar)
  Dilute the ingredients in water (pH should be about 7,3)
  Fill with H2O up till 1000 ml

There are some additional microbiological media used for cultivation of luminescent bacteria:

MICROBIOLOGICAL MEDIA FOR CULTIVATION OF LUMINESCENT BACTERIA


The cultivation should be conducted in temperature 18 - 22C. Liquid cultures should be well aerated (the aeration pumps may be used or cultures should be shaken well). It is necessary to support sufficient amount of oxygen needed in luminescence reactions.

1.
2.

The photos above show liquid culture of luminescent bacterium - Photobacterium luciferum.
When we observe the culture of luminous bacteria in well lit place the light emitted by bacteria is not visible (photo 1). If we observe the same culture in the darkness the intensity of glowing is unbelievable (photo 2). Some of my friends connect this green light with UFOs.
It is interesting to point out that in order to take the photos (particularly photo 2) I had to support constant movement of the broth. If the culture was not mixed the intensity of growing was dropping drastically in some seconds - probably it was caused by lack of oxygen. I solved the problem by using magnetic stirrer.
(click on the photo to see it better)

On solid medium the most intensive glowing can be observed 1 - 5 days after inoculation but faint glowing can be observed longer (even some weeks). Big differences between different species of luminescent bacteria can be observed here (from my observation it occured that the shortest time of glowing have the luminescent bacterium - Vibrio harveyi).

1.
2.

Both pictures show the same culture of luminescent bacterium - Vibrio fischeri on the petri plates with medium "BOSS". Photo 1 was taken with regular light whereas photo 2 I took in darkness giving it long time of exposure.
(click on the photo to see it better)

The above photos show cultures of luminescent bacterium - Photobacterium phosphoreum on petri plates with medium "BOSS". The cultures was started soon before Christmas'97. As you can see we can obtain interesting effects streaking bacteria in such a way that after incubation we obtain glowing pictures.

MORE GLOWING PICTURES


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These pages were created by Piotr Madanecki
e-mail: pmad@eniac.farmacja.amg.gda.pl