I would like to select spore forming anaerobic bacteria by heat treatment. Can anyone tell me how long and at which temperature i shall heat liquid culture to select spore forming bacteria and kill other non spore forming bacteria.
For Bacillus bacteria, I usually incubate bacterial suspension at 80 C in water bath for 10 -15 min. You then can incubate your spread plates in the anaerobic chambers to allow bacteria colonies grow.
Dear Madan,
I may add some points about the procedure since I did some experiments with that. SOmetimes it is important to heat ALL parts of the tube/bottle at high temperature. So, if you use water bath, please, take care that complete flask with bacterial suspension is heated. I used for this tubes with tight screw caps + rubber stoppers and placed tubes completely under water. If you can use an incubator/oven (that full flask is inside it) for pasteurisation it is also good.
Another important point is time of incubation. It takes several minutes to heat the flask with bacterial suspension up to 80C. IN water bath it is shorter, in incubator - longer. I normally increased time of incubation for 10-15 minutes.
Additional important point is that your target - spore-forming culture should be in the sporulating phase. You may easily observe if there are spores visible under phase-contrast microscope.
So, I normally did pasteurisation at 70C, incubation - 30 min only if I was sure that spores were present in the culutre.
Best regards, Alexander.
I worked several years at Konstanz University and enjoyed it a lot! I hope you do too!
Hi Alexander Galushko,
Thank you for your detailed response. Last week performed it. I heated bottle in water bath at 80C for 30 min. Then i was not sure if i really killed non spore sporing bacteria. then i again repeat it for 30 min. Then i inoculated in the fresh medium. This week i could see a growth. But i now i will have to check who are they.
Oh thats nice. I am also enjoying working here in konstanz. :) I think you know our group. I am working with prof. schink.
Best regards
Madan
Yes, I was in Schink Lab. Conditions of work were excellent! Prof. Schink has always interesting projects!
I also liked to bike near BodenSee. There are so beautiful biking tours!
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