We have some sludge from a running SBR reactor and want to culture it under anaerobic conditions for some time prior to inoculation for our research. Unfortunately, our laboratory has no anaerobic chamber or jar, does anyone has an idea how to simply but efficiently create favourable anaerobic conditions to culture this sludge.
Hi,
you could use Thioglycolate broth or a candle jar to grow anaerobic microbes.
you may also have look here:
https://www.ncbi.nlm.nih.gov/pubmed/23867675
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5345310/
We took a simple apprioach.... we used a 4 liter jar and then inerted it with carbon dioxide, and put the lid on. As we were working with a non-sterile feed stock under non-sterile conditions.... it worked just fine.
Thank you all for your suggestions, really appreciate them.
Edward Calt, a friend is using a similar approach, where he put sludge in a 250ml conical flask (tightly covered) and fed it with 0.03g of glucose daily to increase the growth rate. And he got really impressive results.
I may add some more info to the message of Yifan. A serum bottle + rubber stopper + flash gas phase with nitrogen gas via syringe needle = perfect possibility to get anoxic conditions with activa sludge.
One of the observations we made was if you are using mixed culture you do not need to inert it. Just feed it and put a lid on. Our two experiments with mixed cultures were for six months each without interting. Everyday the process discharged enough biogas to inert itseld.
We use customized feeding bottles similar to used in reactor feeding for anaerobic cultivation (One tube dipped in liquid and another exhaust tube attached to a vent filter). Nitrogen purging is required prior to inoculation and intermittently during sampling. If you are using it only for inoculum preparation, one time purging is sufficient. If you are working with non sterile culture, PP centrifuge bottles can also be customized (cheaper option).
I think nitrogen purging to maintain anoxic conditions, we considering nitrogen purging prior to inoculation after culturing.
Nitrogen pirging is required for decreasing the dissolved oxygen content in the aqueous phase. In this case N2 substitute it via bubbling. The vigorous stirring (shaking) helps a lot to speed up the process. In such a case it is possibel to decrease the level of dissolved oxygen to the comfortable level for growth of denitrifyers, Fe(3+) reducers SRB and others.
There was also a couple of publications about using E coli to remove oxygen from media for cultivation of strict anaerobes.
@ Edward Calt, with your observations with mixed cultures, what did your team use to feed the culture and how did you control the biogas produce over that long?
Does anyone has some experience with culturing anaerobic microbes to be tolerant in high salt concentrations, as stated in this article.
Guo Jian-Bo et al., Decolorization of azo dyes with high salt concentration by salt-tolerant mixed culture under anaerobic conditions, J. Environ. Sci., 17 , 2005
We used food waste as a feed stock. With a mixed culture there are some aerobic microbes that will consume the residual oxygen. These anaerobes are supportive to the mixed culture.
What do you mean by high salt concentrations? Look to the environment where the anaerobic culture was derived. This will provide an indication of the required level of salt for that culture.
Great, food waste as a carbon source right?I am feeding my culture with glucose but was thinking of other alternatives.
And about the high salt concentrations, I want the microbes to adapt to high salt environment because after culturing I will be inoculate them in saline environment.
Yes, food waste as a carbon source. Adding glucose will probably accelerate the process. You may need to go through a transition phase from the natural to the new environment.
What is the purpose of the saline environment?
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