Anaerobic bacteria - (Apr/12/2012 )

Hi,

I would like to know if it´s possible to grow a strict anaerobic bacteria (C. sphenoides) in a liquid medium (in a plastic sterile tube) within an anaerobic jar.

Thanks,
vanu

vanu on Thu Apr 12 16:13:46 2012 said:

Yes, why not?
As long as you make sure there is no O2 present.
Its possible you need to flush it with N2.

But I can´t flush it with N2, I don´t have what it is needed in the lab. My problem is that I have liophylized pure bacteria, and that I´d like to grow them in a liquid medium so I could make aliquots (20% of glycerol) and then freeze them, but for the anaerobic bacteria the only thing that I have is the anaerobic jar, so I thought that I was only able to resuspend and inoculate them in an agar plate and the introduce these plates in the anaerobic jar. But what about the liquid medium? I won´t be able to do the same thing without N2?what do you recommend me to do?

Thanks,

vanu on Fri Apr 13 06:23:21 2012 said:

Its not perfect but you could give it a try.
Use tubes that enable gas exchange , so it would have the same effect as the plates really.
Anyway, in the tubes itself, at the bottom you could get anaerobic growht anyway.

Ok, thank u very much.

Hope this will work. And just another question, once a bacteria´s incubation period has passed, for example 24h at 37 degrees for an E.coli, would it be wrong if i leave the plates at 37 degrees for 48 hours more?

Thanks for your help

vanu on Fri Apr 13 07:52:21 2012 said:

"wrong"... its not "wrong" per se. But the bacteria would die in the end.. so you would have less viable cultures.
But the main issue with the "too long" part is that you could have problems with plasmids or leaky bacteria: meaning that some bacteria that have the possibility to break down an antibiotic, could start to break down too much (leaking proteins) and that some non resistent bacteria could start to grow.
Also they start to overgrow eachother, so you cant really use those plates to count.

So its always best to stick to the regular time... but if you really cant take them out.... there is no real harm in it as long as you know what you are doing... (depending on the situation, what type of bacteria, etc..)

PS. also: if you want to continue to work with those bacteria, that stayed in too long, its best to make a fresh culture first from the old one and then continue.. so dont start your experiment with the ones that stayed in too long.

Thank u very much!

You could grow it in properly-prepared thioglycollate. In any case, I encourage you to experiment rather than waiting for anwers from the net - try it. Presumably, you can see growth and in any case you say you've cultured it under anaerobic conditions on plating media -so you should control each step up and through culturability as a lyophiliized material.

As pito said manitaining at 37C is not worng - but the plates will dry out more rapidly.