Dimitri Biological Lights: Science Fair 2013

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.

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.

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

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.

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

Step one- get a fish

Success! It’s glowing!

Oh- and it stinks!

Making gator-ade petri dishes

No glowing colonies on the plates-

• Does this even grow in gatorade?

• Nope

Determining which sea-life grows Fisherii best

What DOES it like?

•Seawater?

•Fish?

•Octopus?

•Squid?

Best: squid

Conclusion

• Don’t believe the literature- test it