The sporulation requirements of this species seem to differ from those of the closely related Lysinibacillus sphaericus. I have tried using a sporulation salt medium published to work with the latter (and with Bacillus thuringiensis); although I was able to induce sporulation in another Bacillus species that I was growing at the same time, I could detect no free endospores in my L. fusiformis culture even after 5 days of shaking at 30degC.
Thank you for any suggestions you might have.
I have about a dozen L. fusiformis strains in my lab. They seem to sporulate reasonably well with our standard sporulation agar, 2xSG (see attached). Please let me know if this doesn't work for you.
Daniel, I attempted the 2xSG agar this week and saw no sporulation by my L. fusiformis isolate after 24 h; the colonies are slightly larger than on my nutrient agar control plate. The other Bacillus species I used as a control, however, did form endospores on this medium that you suggested; no colonies were observed at all.
I also checked the cells by phase microscopy for the presence of endospores, just in case.
I would recommend incubating the cultures at 30°C for 5-6 days and then checking for spore formation.
Thanks for the suggestion Daniel, I took a look again after 48 h and saw that the majority of the L. fusiformis cells on 2xSG agar now carry endospores. Thanks for all your help.
Dear Dr. Mark, I hope you are very well.
I am interested in obtaining spores of strains of Lysinibacillus sp, as part of the development of my doctoral thesis. Although I have been able to induce the production of endospores, the yield is low, I would like to know if you could find an adequate medium for the production of endospores of this bacterial genus.
Any support will be very useful to me.
Regards!!
Hi Manuel, I'm sorry that I am only seeing your message now.
However, I have not had success getting the L. fusiformis strain that we isolated to achieve better than ~60-70% sporulation. I did request another L. fusiformis strain that sporulates more readily; I think my student reported ~80-90% sporulation on 2xSG agar after a few days.
Did you ever find a more efficient method of sporulation for your Lysinibacillus sp.?
Dear Dr. Chee, I hope you are very well. Excuse me please because so far I was able to read the message.
Apparently, the effectiveness of sporulation depends on each strain. Out of 12 strains of Lysinibacillus sp, I only got a very good response from 4 strains in solid Donn Kenny medium (although I tested several, Donn Kenny solid was the most effective). I consider important the fact that of two strains of the same species, for example, L. sphaericus, one showed good sporulation, while the other did not. With the strains that did not sporulate well, I could not work with spore solution, I had to do vegetative cell suspension.
Manuel, I apologize for my own delayed reply. The past semester has kept me busy. Your findings seem to agree with my own poor experience sporulating the L. fusiformis strain that we isolated.
Do you have more information concerning Donn Kenny medium? I was unable to find this information after trying a casual search.
Thank you!
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