it doesn't have to be dreadful

Dilworth dilworth at megsinet.net
Fri Feb 11 17:04:22 EST 2000


First of all, I'm NOT in research.  With that out of the way, my
impression
with pseudomonads is that they don't grow well or at all anaerobically. 
We use
a Pseudomonas aeruginosa control plate in our anaerobic chamber.  If
there is
good growth on it the next day (we sub it daily) we know that air got
into our
chamber.

Just throwing that out to confuse matters.......I'm not sure if Ps.
stutzeri is
similar in its requirement for oxygen.  Are your caps on loosely on the
aerobic
cultures (assuming you're using broth?)  What about CO2?  We incubate
respiratory and wound specimens (likely sources for pseudomonads) in a
CO2
incubator with at least 5% concentration.

Judy Dilworth, M.T. (ASCP)
Microbiology 15 years (Clinical)

Danielle Harvey wrote:

> I, also, would love to see more microbio being shared and talked about
> here.  I am new to the newsgroup by 1 month and have yet to see any string
> of news come along much related to microbio.  So here goes, if anyone is
> waiting for someone to start...
>
> I am working with P. stutzeri, trying to see expression of nitrate
> reductase.  I am growing it in anaerobic minimal media with a nitrate
> headspace trying to encourage denitrification.  I am also growing it in
> aerobic minimal media.  Unfortunately, it is not growing well under either
> conditions.  I have tried 2 slightly different minimal media's, including
> Tanner's.  Basically, I have a nice happy culture growing in LB with off
> the scale optical densities.  Then I centrifuge the cultures, pellet the
> cells, wash with minimal media and resuspend in minimal media.  I expect
> the long lag phase I've been getting since the bugs are in a less rich
> media, but then they won't double beyond the first time.  I know they are
> still alive for the week that I moniter before giving up and that they are
> remembering their rich LB home for that first doubling.  I've tried various
>
> concentrations of vitamins and trace metals and now am at a loss as to how
> to successfully grow them.  Paper's have been published with P. stutzeri
> growm in minimal media with & w/out O2 so I know it can be done.  Any
> ideas?
>
> Danielle
>
> ---




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