I need to prepare spore suspension of Bacillus spp. isolate. How to do this using a simple technique?
In a very short way and without application any additional techniques, let the bacillus cultures continue to grow till the decline phase of growth (3-5 Days) in this case you will get the spoes.
It is known that stress may induce the formation of spores in microorganisms. You may try oxidative stress to induce formation of spores in your culture
In Bacillus, sporulation is induced by nutrient depletion. When cultures reach stationary phase, if there no longer repressing levels of carbon, nitrogen, or phosphorous sources in the medium, they initiate sporulation. Many species also require certain ions in the medium at low levels, especially manganese, magnesium, and iron. Some isolates will sporulate reasonably well on Nutrient agar or on TSA. However, it is best to use a specialized sporulation medium for many Bacillus species. Does your isolate belong to the Bacillus subtilis group (B. subtilis, B. licheniformis, B. amyloliquefaciens, and so on)? Then I would recommend 2XSG medium. There are different media for the B. cereus group. Please feel free to contact me with more information about your isolate; I may be able to recommend media for you to try.
Hello,
Adam, it was smiple and easy to apply the method you suggested. I tried it and I got satisfactory results. Thank you for the references too.
Daniel, yes my isolate belongs to the B.subtilis Group. Can I know why I should use a medium specified for sporulation like 2XSG meduim instead of applying a simple technique such as heating at 80 degree Centigrade?
Thank you all :)
Heating at 80C will not induce sporulation. Instead, it will select for spores that already exist. Vegetative cells from the B. subtilis group will be killed by 15 min at 80C. They will not go on to form spores. However, if any spores already exist, they will survive the heat shock. At that point they will readily germinate if provide with fresh complete medium.
I agree with Daniel. This method is the best one. I tried It many times ago to prepare Bacillus suspension and It worked. Hé à tin will only kill vegetative cells or some times indice spore germination if you want so.
Regards
Please have a look at the following RG discussion threads.
https://www.researchgate.net/post/How_to_sporulate_the_bacteria_or_how_to_induce_the_sporulation_in_bacteria_especially_for_Bacillus_spp
https://www.researchgate.net/post/How_to_get_spores_from_B_cereus_B_licheniformis_and_B_stearothermophilus
Please also take a look at the following RG link and PDF attachments.
Article Induction of Bacillus subtilis sporulation by nucleosides: I...
Adam Ostrowski
Sir, after preparing the dilutions and incubation at 80 degree centigrade, what steps should to be followed to sustain the bacillus culture in spore form?- Badges
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