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Ubriaco
11/02/06 06:42 PM  
other bugs and more questions for Al B
Sorry for picking your brain Al.

The Orval dregs post has me thinking. I have been culturing yeast crudely for a while now, and just started on monday culturing in the lab. So my buddy who was a yeast geneticist in a former life is the guy who started me down this path (I'm a plant molecular geneticist, and he kept telling me to "just do it right, you already have the know-how"), and his protocol for -80 stocks is 300 uL saturated culture and 700 uL 75% glycerol. He specifically said that this works only for S.c. since they can only handle the extreme osmotic pressure that places on the cells, due to their evolution for handling the osmotic pressure of alcoholic fermentation.

So the question is if I were to culture from a bottle of Orval, or tried to culture my lambic culture would they be able to handle that level of stress, since they are evolved for growing in these conditions as well? I am really proud of my lambic culture since it consists of the dregs of every bottle of unpasteurized lambic I have ever drank (of which there are many).

A side note: I was looking at my cultures under the scope yesterday and to my amazement found only uniform S.c., of which about 75% were in the process of division. So I was really happy to see that. My old method of culturing consisted of blowing up cultures in my reactor, allowing for flocculation, and then placing the cake in Ball jars in my fridge. Periodically I would repeat this to ensure viability. That method, although it really worked for me for about 2 years, was a recipe for an infection. So it seems that I started this just in time.

Also if the answer is that they cannot handle that stress what would they like better, and is there a formula for plates and liquid media that would be better suited for pedio, brett, etc?

Sorry for all the questions, but I really want a comprehensive library of all the yeast I use and some I may never use (for friends).

thanks

Al B
11/03/06 07:47 AM  
Re: other bugs and more questions for Al B
Maintaining your specific cultures is great (I've done it in past too). << and his protocol for -80 stocks is 300 uL saturated culture and 700 uL 75% glycerol. He specifically said that this works only for S.c. since they can only handle the extreme osmotic pressure that places on the cells, due to their evolution for handling the osmotic pressure of alcoholic fermentation. >>

- well his theory seems fine and he's probably right, but I don't see it necessary. Freezing cultures, including eukaryotic yeast cells do just fine with a final concentration of 15% glycerol. As you know, the glycerol protects the cell walls from rupturing during freezing and thawing. After thawing, the culture should be added to media and incubated (Going from 50% glycerol to a starter gravity may be osmotically challenging too).

One note, if its a mixed culture, different bugs will grow at different rates so your starter may get wierd.

Folks who don't have a subzero freezer can maintian bugs in the refridge on certain types of modified wort medias.

Pedio seems to do better in a liquid base fortified with tomato juice and other stuff (I can get you the recipe)-but you need an anaerobic environment. Bretts should have 0.5-2% CaCO3 forlong term storage.

cheers

Al Bactrometer

Ubriaco
11/03/06 08:49 AM  
Re: other bugs and more questions for Al B
Thanks Al,

good to know

 
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