I recommended yor question to other to answer

Please read over here: http://www.surgeryencyclopedia.com/A-Ce/Anaerobic-Bacteria-Culture.html

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2687301/

Azadeh, your description is without experimental details - not easy to find the reason why your SRB do not grow. WHat kinds of SRB? Very often SRB complete oxidizers do not like to be cultivated in  anoxic chamber and also in anoxic jars.

Alexander, I am growing Desulfovibrio species. I could grow them in my third trial however my sample was contaminated with another bacteria. Do you know how I can separate the Desulfovibrio?

Azadeh, normally Desulfovibrio grow well in defined medium with lactate as the only organic electron donor. If you have in your medium other organic compounds you need to exclude them and make "agar dilution" procedure.

But !!! you should not do it in "anoxic chamber".Very often that is the source of contamination of pure cultures!!! You can do that on a lab bench with fire but you only need to be rather quck not to oxidise your medium!!!

Alexander, I am a bit confused, so I should not do the culturing in anaerobic chamber? don't the bacteria die on the bench? That would be great if I can do the experiment outside the chamber, cause the one we have at university is highly contaminated and it's not practical to sterilize. 

I received vials with freeze-dried bacteria from ATCC. Can I rehydrate the pellets on bench?Isn't it difficult, cause as soon as as I open media bottle O2 starts diffusing. I can bubble N2 on top of my media bottle but I cannot do that on top of small glass vials.

Dear Azadeh,

I was away from the office for quite long and could not answer your question.

I normally re-hydrated the cultures pellets from DSMZ on the bench. For this I put required amount of liquid medium supplied with a reducing agent OR just following instructions. For this purpose I use syringe with long & thick needle! Later I took the suspended pellet into the syringe trying to get no big air bubbles into the syringe (and remove the bubbles from the syringe - by pressing them out via the needle. And I put this inoculum into the tube with anoxoc medium that has inert gas phase and sealed with black rubber stoppers.

With best wishes, Alexander.

Azadeh: even if the paper is old one, the procedure described for Desulfovibrio cultivation is simple and works perfectly!! Please see the following link:

Article Growth of Desulfovibrio on the Surface of Agar Media

Good Luck and Best regards

Vit

Vit, thank you for this paper. It is a good hint for Azadeh"s work! It is nice to recover such knowledge and use it!

Some modern explanation for the results of cultivation experiments.

Most of Desulfovibrio spp. are not autotrophs and to grow better with hydrogen in the mineral salt medium they need to have acetate + CO2 for assimilation (addition of yeast extract is beneficial for growth since it supplies organic that may be assimilated + additionally "well prepared" yeast extract has also reducing power = may serve as soft organic reducing agent).

Stimulation of growth of Desulfovibrio on mineral medium in the presence of TSA plus salts medium with grown Desulfovibrio might be explained by acetate volatilisation. TSA based medium was not buffered and acidification caused to increase volatility of acetate formed from lactate as acetic acid.

TSA plus salts medium looks a bit as Posgate Medium B = media that has sufficient amount of Fe2+ to detect sulfide-prodicung colonies as black color producers.

However, I would to warn that blackening in the rich organic medium might be caused by not only SRB BUT but by bacteria fermenting sulfur-containing organics!!! Bacteria release sulfide as a by-product during fermentation of such organics. So, it is important to have really pure culture of Desulfovibrio.

Hi, Does anyone have a pdf copy of the above article?