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Matt B
12/19/07 10:44 AM  
Making slants of "bugs"
I'm currently making slants of any yeast that I use in order to build a local yeast library (and to save $$). I'm planning to start doing some funky beers next year and I have a question about slanting up the bugs. I realize that it would be difficult to slant up one of the mixes that have pedio/lacto/sacc/brett and be able to get an even mix of all cultures but is there any trick to slanting up one of the pure cultures from White Labs or WYeast on the individual strains?

Thanks in advance!

~Matt

Al B
12/19/07 10:56 AM  
Re: Making slants of
For growing yeast on an agar slants, remember to cap the tube loosely so that O2 is available. After 3-7 days, tighten the cap and store refrigerated.....optimum temp. as close to 32-34F is best for long term. Also, after 1 year or so, you should subculture onto fresh agar again and so on since they won't keep indefinately under these conditions.

Matt B
12/19/07 11:03 AM  
Re: Making slants of
Right, I'm already doing that with standard beer yeast. My question was if there are any other steps I should take to do the "bugs"? Like do I wait longer before I put them in the fridge or should they be reslanted sooner than standard yeasts. Aren't some of these stressed in the presence of O2? Should I purge the vials w/ CO2 before slanting them?
Al B
12/19/07 11:28 AM  
Re: Making slants of
Oh right.....the bugs.

Treat the Brett. like Sacch. yeasts. I do not feel they need to be re-slanted sooner than other yeasts, but all strains are different - you'll have to monitor them.

When I say 02 in this case, I meant air. Lactobacilli are microaerophilic (tolerate air, some strains will grow lightly such as L. delbrueckii), while Pediocococcus will not grow at all in the presence of air. It is best to culture Pedio in a broth for this reason (although one can use CO2 - this is still tricky).In addition, Pedio requires specific nutrients, so yeast nutrients are a must.

Matt B
12/19/07 11:32 AM  
Re: Making slants of
So for Pedio maybe make a slant that is all wort w/ nutrients mixed in with it instead of using agar and maybe rig up something that will put off CO2 (like an extension from my keggerator system on < 1 psi)to purge the tube after I've innoculated it?
Matt B
12/19/07 11:39 AM  
Re: Making slants of
Or should I just be more "traditional" and do the Vinnie C. method and keep my cultures of Pedio and lacto on some oak chips or cubes? :)

I may do that in addition to the slants anyway.

Al B
12/19/07 11:44 AM  
Re: Making slants of
OK. But even better would be to "stab" the slant so that the Pedio is down deep inside the agar. This will be a bit of a pain to reculture, but will ensure viability of a fastidious anaerobe such as Pedio. Also, in years past, microbiologists used sterile mineral oil to overlay the slant cultures for storage. Although I don't recommend this complexity, you can see how anaerobes differ from other bugs. I would do a stab, rather than a slant for Pedio.

Matt B
12/19/07 08:26 PM  
Re: Making slants of
How would you go about reculturing a slant from stabbing it?
Al B
12/19/07 10:31 PM  
Re: Making slants of
Do you mean how you would stab a slant or how to subculture from a stab? You can use an inoculating loop to stab inside the agar where the growth is. This may break up some of the agar, but its the easiest way to get the cells. You may want to have a couple of these tubes as a back-up for more subcultures, but I still feel the easiest and best way is to use a broth for Pedio. Then you can soak the chips too. Good luck....

Al Bacteremia

Matt B
12/22/07 05:36 PM  
Re: Making slants of
By making a broth do I just use wort or is there other ingredients? Do you have a "recipe"? I appreciate the advice by the way. I did a quick google search on "Fastidious anaerobe broth" but it looks like all it comes up with is commercially available medium for purchase. I'm sure there is a way to make this at home.
 
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