Source: https://m.carolina.com/teacher-resources/Interactive/living-organism-care-guide-vibrio-fischeri/tr23902.tr
Timestamp: 2019-04-22 18:08:43+00:00

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Vibrio fischeri cultures must be subcultured 2 to 3 times per week to maintain their bioluminescent glow.
Bioluminescence is observed most readily when cultures are 18 to 24 hours old.
Observe Vibrio fischeri cultures in a dark room completely devoid of light. You should allow your eyes to adjust to complete darkness for at least 5 minutes before observing cultures.
Vibrio fischeri is a motile, rod-shaped marine bacterium that produces bioluminescence.
It is frequently found living in a symbiotic relationship with marine animals.
The expression of bioluminescence is controlled by a small set of genes known as the lux operon.
The genome of Vibrio fischeri is contained on 2 chromosomes.
We ship V. fischeri as a tube culture or a plate culture .
Culture requirements for V. fischeri are more demanding than other familiar bacteria such as Escherichia coli.
To maintain bioluminescence, subculture V. fischeri at least twice weekly (3 times is better).
Take the culture into a completely dark room and wait for your eyes to adjust until you can see the glow (at least 5 minutes).
No housing information applies for this organism.
No feeding information applies for this organism.
Culture V. fischeri on photobacterium agar or photobacterium broth. Subcultures should be made from the glowing areas of the culture and inoculum should be transferred to a petri dish or tube containing photobacterium agar.
Note that V. fischeri should be inoculated more heavily than other bacteria.
Carolina also sells plates and slants of photobacterium agar for your convenience.
After making the subcultures, clean the work area with disinfectant and wash your hands again. Following inoculation, incubate the plates or tubes, with caps loosened, at room temperature (20 to 26° C) and in total darkness for 18 to 24 hours before observing for bioluminescence.
When you have finished setting up the cultures, clean the work area with disinfectant and wash your hands again.
To dispose of cultures, autoclave them at 121° C for 20 minutes or submerge them in a disinfectant such as 70% alcohol or a 10% bleach solution overnight. Cultures can then be discarded with regular solid waste.
Vibrio fischeri is classified as a biosafety level 1 organism. Before you work with bacterial cultures, wash your hands with soap and water, ensure that the work area is draft free, and wipe down the work surface with 70% alcohol or a similar disinfectant. Never work in an area where food is prepared or consumed.
What agar should I use for my V. fischeri cultures?
V. fischeri is a marine bacterium and needs salt in its growth medium. We recommend photobacterium agar or photobacterium broth for best results. V. fischeri will not grow on nutrient agar and other common media.
Can I pour my own plates if I don’t have an autoclave?
See our photobacterium agar. You can pour 4 to 5 standard 100 x 15-mm plates with 125 mL of medium.
Why am I not seeing a glow?
Oxygen is required. Be certain the caps are loosened on the tubes.
Check the age of the cultures. Bioluminescence is most visible 18 to 24 hours after inoculation.
Observe your cultures in a totally dark room—curtains or blinds over windows are not enough. Give your eyes time to adjust; the glow is faint.
Check your medium, as V. fischeri will not glow unless you use a medium specifically formulated for bioluminescent bacteria.

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