Vibrio harveyi (Johnson and Shunk) Baumann et al.

1. Open vial according to enclosed instructions.
2. Using a single tube of #101 broth (5 to 6 mL), withdraw approximately 0.5 to 1.0 mL with a Pasteur or 1.0 mL pipette. Rehydrate the entire pellet.
3. Aseptically transfer this aliquot back into the broth tube. Mix well.
4. Use several drops of the suspension to inoculate a #101 agar slant and/or plate.
5. Incubate the tubes and plate at 26°C for 24 to 48 hours.

This strain will grow in #2 Marine Broth 2216 (BD 279110) and on #2 Marine Agar 2216 (BD 212185), but Photobacterium Medium is required for demonstrating luminescence.

To check for luminescence, inoculate a #101 slant, leaving cap loose. Incubate at optimal temperature until growth is observed. Then, hold in the dark for 10 minutes. If no luminescence is detected, reincubate and check again after 24 to 48 additional hours.

When maximum luminescence is obtained, close stopper tightly. Demonstration of luminescence requires that vial be rehydrated with #101 broth. Transfers from #101 broth to #2 broth may yield more growth.

Colonies on #101 agar are circular, entire, glistening, and smooth.

Additional information on this culture is available on the ATCC^® web site at www.atcc.org.

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