The document outlines a plan to demonstrate the properties of fluorescent bacteria, specifically Vibrio fischeri, by growing them using a mixture of fish, Gatorade, and gelatin. It provides detailed equipment and preparation instructions, emphasizing the safety measures needed due to the bacteria's relation to harmful strains like Vibrio cholerae. The project aims to cultivate glowing colonies in a high salt environment to ensure the survival of the desired bacteria while preventing contamination from harmful strains.

1. Biological Lights
By Dimitri Zlatev

2. Background
• Fluorescent bacteria is abundant in nature
• Ocean ships often report glowing trails in their wake
• Some tropical beaches have glowing waves at night
• Last year, contaminated meat in Sichuan, China glowed blue.

3. My Plan:
• Demonstrate the properties of fluorescent bacteria.
• Bacteria is only avaliable to high security labs
(so it’s relitatively hard to get).
• Grow fluorescent bacteria from a fish, some
gatorade, and some jello.

4. Background and references
• A Mohankumar*& P Ranjitha: Purification and
characterization of esterase from marine Vibrio
fischeri isolated from squid. Indian Journal of
Marine Sciences Vol. 39 (2), June 2010, pp.
262-269.
• Peter Herring: Marine microlights: the luminous
marine bacteria. Microbiology
TodayVol29/NOV02

5. Equipment
• Pressure cooker
• Glass petrie dishes
• Blender
• Incubator for smelly fish plates
• Wooden coffee stirrers

6. Supplies
• Gelatin
• Gatorade
• FRESH ocean fish or squid
• Rubbing alcohol

7. Plate Preparation
• Use the recipie for knox blox (2X concentrated gelatin) on commercially
available gelatin projects.
• Pressure cooker to sterilize equipment.
• Assume gatorade is delivered sterile, to be added without further boiling.
• Mix gatorade jello (2X) and pour into sterile glass petrie dishes. Need at
least 10 plates.
• All plates to be bagged and refrigerated until use.

8. Fish Prep
• According to the Herring, bioluminescent bacteria are leaking out of
creatures all the time. You COULD culture them out of sea water, but it is
possibly easier to gather them from a squid
• Puree the squid, plate, and store in humid warm dark place.
• After 48 hours, sit in the dark with the plate for 15 minutes, then pick
anything that LOOKS glowing, and transfer it to a fresh plate. You are
selecting for glowing colonies, so they can multiply.
• After 4 or 5 transfers you should see glowing colonies

9. Dangers!
• We intend to grow Vibrio fischeri, a waterborne ocean
bacteria.
• Its cousin is Vibrio cholerae, a freshwater contaminant
that kills more than 100,000 people per year.

10. Public Health
• So how come our stinky fish doesn’t cause
a plague in San Francisco?

11. SALT!
• Gatorade Endurance Formula is a high
salt formulation. Fisheri loves it, cholerae
cannot stand it, so we are selecting for
bacteria that can endure a high salt
concentration.

12. Have FUN!
• I mean, how cool is that?
• When is the science fair????