I need help on Isolating Vibrio Fischeri from squid and fish

[JimmyNoir]

so I'm working on a project where Im trying to isolate Vibrio Fischeri from squid and fish, for the squid I put it in a tupperware with saltwater then placed it over some ice in a cooler in a dark room and as for the fish I cut the skin and put it on a plate with saltwater then put it over some ice in a cooler in a dark room but all I have seen for now is a small very tiny glow, am I doing something wrong? does it really need to be cold? 

[Nathan McCorkle]

See here:
https://web.archive.org/web/20110111111314/http://has100ideas.com/blog/culturing-bioluminescent-microbes-part-1

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-Nathan

[Mega [Andreas Stuermer]]

I once isolated them at basement temperature but I afterwards was told this can lead to Vibrio Cholerae growing.

[Mega [Andreas Stuermer]]

Good news, fellow basement biotechnologists, 

http://phys.org/news154708942.html

It is stated that active Vibrio Cholerae growth was detectable when the water temperature rose above 20°C 

"All of the three vibrios were demonstrated in Norwegian mussels (at fewer than 100 bacteria /gram) and in Norwegian seawater (up to 30,000/litre) during the course of the study. They were first and foremost demonstrable when the water temperature rose above 20°C.

Read more at: http://phys.org/news154708942.html#jCp"

So, if you leave your fish in the basement (if not heated, so ~10-15°C in winter), Vibrio Cholerae shouldn't grow much and may be outcompetet by other bacteria. Of course it will still be safer to incubate it in the fridge.  

[Mega [Andreas Stuermer]]

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC242147/

"Effects of salinity and temperature on extended survival of V. cholerae demonstrated that, at an estuarine salinity (25%) and a temperature of 10 degrees C, V. cholerae survived (i.e., was culturable) for less than 4 days. "

[Cathal Garvey]

Cholera's more or less the whole reason it's recommended to grow at 4C.
Yea, it won't be happy below 20C, but for the day or two of a difference
it makes to Vibrio phosphoreum (the usual guy you're aiming to culture)
it's not much.

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[Mega [Andreas Stuermer]]

 > but for the day or two of a difference 
it makes to Vibrio phosphoreum (the usual guy you're aiming to culture) 
it's not much. 

Sorry, I don't quite get the meaning. (German speaker :P )

You mean Vibrio Cholerae will grow slowly, but it will grow alonngside V.Phosphoreum+Vibrio Fischeri at 10°C?