Growth of bacteria in controlled atmosphere

Lynn Bry aj088 at FreeNet.Carleton.CA
Wed Jul 26 18:50:30 EST 1995


R.S. Perry (rsperry at bc.ic.ac.uk) writes:
> I am attempting to grow isolate Gallionella ferruginea. The procedure is
> lengthy requiring many serial dilutions.  The situation is further
> complicated by the fact that growth tubes must be kept in an atmosphere of
> 94%N2, 5%CO2 and 1%O2.  I can think of ways of achieving this but does anyone
> have any practical experience of achieving such.  If so I would be very
> grateful if they could e-mail me informing me of their method.  I do have
> access to a Gallenkamp CO2 incubator but this only seems to allow CO2
> enrichment. 
> 	Thanks in advance
> 		Robin (rsperry at ic.ac.uk)


We use similar conditions for growing Helicobacter pylori in a
Campylobacter incubator. The incubator has a small port/hole in the
back of it, big enough to allow an electrical cord into the incubator.
On the bottom shelf we run a small rotating table (for cell culture
incubators, sold by Fischer Scientific ~$300 US) for incubating
cultures. The cultures can be grown either in petrie dishes or
in cell culture flasks kept on the rotating table. Both allow efficient
exchange with the atmosphere of the incubator.

The atmosphere maintained is microaerophilic, pretty much the
same as what you mention for Gallionella. For making serial
diultions all we do is pre-incubate the broth to equilibrate
it with the atmosphere, then do the dilutions as needed. We do them
on the open benchtop, but you could just as easily do it under a 
gas canula of the necessary gas mixture if you don't want to expose
your cultures to atmosphereic O2.

ln



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