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Lambic Digest #1049
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Subject: Lambic Digest #1049 (March 23, 1997)
Lambic Digest #1049 Sun 23 March 1997
Forum on Lambic Beers (and other Belgian beer styles)
Mike Sharp, Digest Coordinator
Contents:
more pedio (Jeremy Bergsman)
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Date: Sat, 22 Mar 1997 00:12:26 -0800
From: Jeremy Bergsman <jeremybb at leland.Stanford.EDU>
Subject: more pedio
In private email Jim Liddil added that he uses amphotericin B in
addition to oxyrase. I believe this is a broad spectrum inhibitor
of fungi. Can anyone suggest a concentration? Sigma suggests
0.3-1 microgram/ml. (It cannot be autoclaved or filtered in
aqueous media. They suggest filtering in DMSO.)
In any case, you should have an anaerobic environment to grow the
pedio.
> From: fespourteille at mmt.com (Francois Espourteille)
> For those of us that haven't reviewed our college microbiology (1982
> or 83?)notes recently, what does the resistance/sensitivity to
> cycloheximide mean?
I'll try to answer. Others may correct if I'm wrong or unclear
due to the late hour.
Cycloheximide prevents the growth of most Eukaryotes (e.g. fungi),
but spares many "wild" yeast. In particular, it spares Brett and
Candida but prevents Saccharomyces growth. In other words, if
it grows on cycloheximide (also known as actidione) it isn't
Sac. and if it came from lambic it is probably brett (or bacterial,
which is easy to tell under a microscope). An easy test that
adds to the certainty that you have brett in this case is to test
for acid production, usually by streaking on plates containing
supersaturated CaCO3 which is milky until it is dissolved by
any secreted acid. Brett will therefore form a clear area around
itself on such plates.
So with ampho and cycloheximide, you can select for brett and pedio
(assuming you are starting with those and sacc.). I have been
doing replica plating to find the sacc.
> From: eurquhar at sfu.ca (Eric Urquhart)
> You could take a portion of the dregs and pitch it into a
> just starting to ferment culture and keep it very warm 30C or better for a
> couple of weeks. Then get rid of the yeast with cycloheximide ( up to 100
> mg/L) which won't kill the pedio or other lactic bacteria.
This is a fascinating suggestion. Do you think the sacc. fermentation
helps by sparging O2 and providing CO2, or by providing fatty acids
and other nutrients (B vitamins?)? I would imagine that this would
starve the pedio for energy sources...? Of course it should be said
to watch out for hops inhibiting pedio growth, although I guess a
few have warned here that you don't want to select for hop-sensitive
pedio during starter production just to have them poop out in the real
thing.
More notes about cycloheximide while we're at it: you can autoclave
it, and fairly concentrated stocks can be made in EtOH. It is pretty
nasty stuff, so handle carefully. I have read that it is inactivated
in dilute alkali, which would suggest that cyc/CaCO3 plates might
not work. Anyone care to comment?
- --
Jeremy Bergsman
jeremybb at leland.stanford.edu
http://www-leland.stanford.edu/~jeremybb
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