Anaerobe bacteriology - C.perfringens (Feb/10/2011 )

hello everybody!
I have a problem, stock C.perfringens cultures growth different anaerobe or facultative anaerobic bacteria since one month, I have used to TSC (with D-cycloserine) agar as a media...

Does anybody have any suggestion supplement or antibiotic etc. for killing the contamination flora,
thank you


P.S. I sent the image of TSC with contaminated.

why not make a fresh stock from a pure colony from your fresh streaked plate?

Regards,
p

rickyvets on Thu Feb 10 09:13:31 2011 said:

Hi, I tried it but the result didnt change:(( I stained the white colony and the colonies appear gram negative rod at microscopy...

rickyvets on Fri Feb 11 08:47:19 2011 said:

Then your stock culture is contaminated. Do you have a stock culture that hasnt been opened?

You could try to make a "new" stock culture, subculering your bacterium untill you get a "clean" not contaminated plate..

pito on Fri Feb 11 11:28:58 2011 said:

Unfortanetly I dont have another clean stock culture:(( there was only one culture... C. perfringens is my PhD thesis but I dont know what to do anymore...

rickyvets on Fri Feb 11 11:44:42 2011 said:

Well try subcultering it.. Take colonies and subculture them... If you have a plate with your bacterium and contaminats, pick your bacterium, inoculate a fresh plate.. and keep doing this till you get what you want.
Or have you tried this before?

There are two types of colonies on the plate -- white and black. Which is the correct phenotype for C. perfringens on the media you used?

There are well-isolated colonies of each phenotype. Pick one of each type and streak it to a fresh plate. Do they each produce a single phenotype, or do they produce both black and white colonies?

yes, black should be the right choice! Take a black colony and make a new stock culture!

or does these colonies appear after streaking a single colonie on a fresh plate? ...in this case i would check if your media is contaminated and do the corresponding controls!

Regards,
p

HomeBrew on Fri Feb 11 12:25:31 2011 said:

I tried all of your suggestions but the result didnt change, C. perfringens growth on TSC as a black colony.
For purification; I tried to take middle of typical colonies via loop with needle so the colonies surronded with white zone. results was same I couldnt get rid of the contamination flora again...
Certainly there is a solution but What? presently, I dont know:(

I will wait your suggestion...

rickyvets on Fri Feb 11 13:50:53 2011 said:

Well then there must be something wrong with your technique... or your incubator is contaminated or the LAF is...

Did you try to pick a black colony with a thootpick? and then subcultivate it?
(just touch a black colony very very gently at the top and place it on another fresh plate)
BTW: have you tried to incubate fresh plates? (with nothing on it, negative control)?

And you are 100% sure its a different bacterium? Because 2 colors doenst always mean its another bacterium.
(but since you said its gram - and C. perfringens is gram + this seems not possible)