I need to cultivate Salmonella Typhi anaerobically with some simple methods. I have already tried growing it in screw cap tubes and sealing the tubes with molten paraffin wax. But i still see some air bubbles after few hours of incubation. Please, help me to overcome this problem by some simple methods.
I used an atmosbag, which is a bag with arms, you put your stuff inside them you close the bag after that you have to fill the bag with nitrogen gas. You also have to bubble your liquid media with N2, you can use a vacutainer or a flask with rubber stopper.
Salmonella are facultative anaerobes, so you could try the simplest method of all - that is incubating the plates in candle jar.
Refer: https://ispub.com/IJMB/4/1/13456
As already mentioned before, a candle will probably do the job. Put a candle in a high beaker, let it consume up the oxigen and put your cultures in. If you have the possibility to degas your liquid cultures with nitrogen gas, do so before incubating them. But even if you don't do that, the organisms will consume the oxygen and start with their anaerobic metabolism.
Greetings
In metal can with plastic cover put your cultivated dished and flame a candle and cover the can . Notice the flame till it end that i mean all oxygen in can is exhusted and the media became anaerobic
use anaerobic poach instead if not jar then press to release excess air in the bag. then heat seal the bag and incubate your culture...we sometimes use the most simplest way of using double bag or even triple plastic bag or more, and it works...anyway, it depends on the oxygen permeability of the film you use...
Salmonella is an anaerobic facultative bacterium but if you need to growth bacteria in anaerobiosis you can use a liquid medium with a high depth. The anaerobic bacteria will growth only in the depth layer but Salmonella will growth in all medium.
Balch et al., 1979 Microbiol. Rev 43: 260-296
Anaerobic technique by Hungate and Balch
Actually you don't need the anaerobic jar to culture your specimen for the growth of S. typhi. Simple culturing on agar like Salmonella-Shigella agar will do the job. Then you can go ahead and do motility test using the stab method. S. typhi is motility positive. The source of your specimen however goes a long way to determine the type of pretreatment before culturing.
Thank you all for your valuable answers.
As I am performing an assay to detect formate ions let out by S.Typhi when grown anaerobically, I strictly have to grown them in liquid culture. Well, I shall try sparging Nitrogen gas if it is available in our institution and will also try putting them inside Plastic bags.
Candle jar is not enough; but you can use a jar (not necessarily anaerobic jar) with an Anaerobic atmosphere generator pouch. Alternatively, you can use thioglicolate broth
Sure, the thioglicolate broth has to be fresh. (Prepared the same day), or to boil it before its uset, as Dr. Friedman says
I need to cultivate clostridium perfringens. Is it possible to use metal can and candle too?
In the absence of the anaerobic jar and gas pack, a canister or a big tin e.g bournvita tin with lid can be used. After you have carefully arranged your inoculated plates inside, you will light a candle and stand it on the plates or any space available in the tin after which you will now cover the tin with the lid and put in the incubator with appropriate temperature and incubate for 24h. However for S. typhi, you don't have to culture anaerobically.
About pyrogallic acid + NaOH as oxygen scavenger ― check my post at: https://www.researchgate.net/post/Can-somebody-recommend-a-method-for-making-48-96-well-plate-anaerobic
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