Hi,
I'm lookong for methods for the isolation of microorganisms from liquid consortium, which could not grow on standard, solidified agar. Strains which grow on agar plates are unable to degrade xenobiotics used by me, also when I combined pure strains in one culture. So I guess I still do not isolated strain from a consortium which is able to use used xenobiotics as carbon and energy sources. Any ideas on how to get this strain? I also know that they exist as viable but nonculturable bacteria but I hope there is a method to isolate some of them after all.
Sir, Have you tried Gelrite (GelzanTM)? I have used it as alternative gelling agent for non-agar culturing conditions such as high temp.
Hello Ariel,
Well, here is my suggestion, just remembering old experiments carried out in the lab in which i amde my PhD a looooong time ago.
Agar was digested by a rare bacterium and we had to susbtitute it with gelatin (same amounts of gelatin per liter of culture as agar).
Maybe it could work to help you to solve your isolation problem?
Best regards,
Philippe Urban
I'm using agarose at a concentration of 1.5% to solidified broth. I thought about using gelatin instead agarose but I was afraid of plates hydrolysis. I made few biochemical analysis on four strains isolated from consortium (strains without drug degradation potential) and three of them shown ability to hydrolysis of gelatin.
Anyway, thanks for the advice.
Sir, Have you tried Gelrite (GelzanTM)? I have used it as alternative gelling agent for non-agar culturing conditions such as high temp.
Hi Ariel
The lysis of gelatin does not mean, that you cannot plate on Gelatin, if there is no other possibility, you will get a corona-forming on the place where you have the gelatine-consumers but you can pick it from there and you have a criteria do discriminate MOs in consumers andn on-consumers
If Gelatin is not Agarose and Gelrite are good alternatives, also any kind of gel-formíng Carbohydrate like Xanthan, Chitosan. Maybe they are more difficult in preparation or solidifying, thats why Agar is so popular.
If you have access to a FACS-Laser Cellsorter, you have also he possibility to isolate single cells from and into liquid cultures, if there is any kind of difficulty in plating anyway.
Good luck
Thanks for advices, I will try use other than agarose substances to solidifying plates. Unfortunately in my department I found only gelatin so for others I must order. I hope this change give some results. I'll probably give some feedback about any results. Thanks again.
Unfortunately, Frank, I don't have possibility to use FACS-Laser Cell sorter, but thanks for the idea anyway!
I used dilution method, of course, in few dilutions of consortium. I'll try use others solidifying, agents, thanks for advices!
I'll see if this give some results.
We have the same problem with an anaerobic strain. They dont like to grow in agar.
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