I am having some difficulty getting the honey isolates dd1-8a, dd5-Q, dd8-63a, dd11-2a,dd12-22a, and the roach trachea isolate to grow from -80C. They grew on MYPGP agar at 37C without an issue, however they are not growing in broth media.
For the second time, the honey isolates did not grow in BHI broth media. Isolated colonies were inoculated into 10 mL of BHI broth media and incubated at 37C shaking at 225 rpm. Even after three days there was no visible growth observed. I have been able to get growth performing this method in the past with these isolates, however that was with BHI broth media I had prepared myself. It is possible that the BHI media I am currently using is from a different source and results in no growth of my isolates.
Regardless, I have inoculated isolated colonies of each of the isolates into 10 mL of LB broth media and incubated them at 37C shaking 225 rpm. If I do not observe growth after 24 hours I will begin to suspect there is an issue with the bacteria themselves and perhaps not an issue with the media.
//EWW
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