I am trying to isolate single colonies from a mixed culture. The colony actually seems single on plates. I did the gram stain and took these results you see in the pictures. I don't how a bacteria should look like when sporulating, could you please tell me if its pure?
Ps: Sorry about the quality of my gram stain, ı am still a newbie!
Well, be sure the bacterium is isolated on its specific medium. Ok you did Gram stain and looks Gram positive. If the bacteria are bacilli or clostridia which are sporulated bacteria ,you can make spore stain. Any how spore stain is necessary. Your Gram stain is well
Dear Prof. Enan, i can't use a spesific medium since i am trying to find and isolate all living bacteria in this mixed culture. My lab does not have malachite green yet so i have to wait for the spore stain. But if it was clostridium, would it be still looking like both rod and tennis rocket shaped when gram stained? Is it possible?
It is correct,but it is difficult to precise from the photo that it is clostridia. If you are working under aerobic conditions , this organism may be belonging to bacilli. Test firstly the identification experiments such as catalase, nitrate reductase.....etc.A simple way is to use Vitek 2 , a soft identification equipment and within 24 h inform you the correct name of the organism via many tests can be carrien thorougly. Also try to get malachite green and make spore chain to declare more correctly.
Your isolate appears to be pure. Refer to the link to know about your tennis racket shape. http://genesdev.cshlp.org/content/19/18/2236.full
I checked the article, thank you so much for your time and answers Mehmood and Bhaskar.
Prof Enan, I will do the tests. What do you think about Dorner's method for endospore staining? Can ı use it instead of staining the culture with malachite green?
Bacterial cell measure one micron in diameter where as fungal cell measure 2 to more microns in diamter..
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Hello Yağmur
The culture is seem a pure
And about the malachite green, you can find useful references on attachment 1
https://microbeonline.com/endospore-staining-principle-procedure-results/
4
https://www.slideshare.net/prashanthkumarguddeti/staining-techniques-in-microbiology
5
https://docs.google.com/document/d/1xt0s7Vcd-rqySbzvm2G4Y5hJDY3GuMXufKGNr0vXcdQ/preview
9
https://www.koofers.com/flashcards/biol-microbiology-exam-i-la/review
15
https://delrio.dcccd.edu/jreynolds/microbiology/2421/lab_manual/spore_stain.pdf
19
https://www.microbiologyinpictures.com/mycobacterium-tuberculosis.php
20
https://www.piwet.pulawy.pl/jvetres/images/stories/pdf/20074/20074695700.pdf
Dr. Al-Ani,
Thank you so much for the resources you've been a big help
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