I am going to run MIC with Aspergillus Niger and Fumigatus. I cultured them in broth, Sabouraud Dextrose Broth, overnight at 37 one time at and 25 the second time, but they form big cotton balls and I cannot get an accurate OD to run MIC using microdilution method. I also spin them down and did vortex to get them dissolved but it did not help. Is there anybody who knows how to fix this problem? Should I add Tween to the broth to make the cotton ball/droplets soluble?
Hello Marjan
You need to change the producerby using the following references, hoping it work better.Good luck
Hi Dr. Al-janabi
Thank you so much for the attached paper. It is a good source for MIC protocol, but it does not talk about how to grow them in broth in a way which avoids forming the cotton balls, however it mentions the right incubation time and temperature.
Mushroom cultivation in a liquid medium always requires agitation for aeration. in your case I think it is necessary to use a rotary incubator , you can test different speed of rotation 150 revolutions per minute, 200 rpm....., with an incubation time not exceeding 24h to 48h. this to promote sporulation and inhibit mycelial growth
Thank you so much for the information. I used 120 rpm last time, I will try 200 rpm to see if it fixes my problem.
Hello Dear Marjan !. In order to find the specific concentration of Aspergillus spps you better culture them in solid medium like YES agar at least for three days at 28-30 degree C. And then you can dilute the spore with sterilized water containing tween 20. In order to determine the spores concentration you can use either SPM or microscope.
Dear Firew, I tried what you suggested and it worked perfectly. Thank you so much!
Dear Marjan,
I have the same coton balls as you. What kind of protocol did you use in the end ?
Thank you
Marc
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