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. -_ @ @ © « FD-302a (Rev. 10-6-95) 279A-WF-222936-USAMRIID Continuation of FD-302 of ee ,On April 26, 20Péee 2 rr or ot a en moO Ba spore growth in flasks with Leighton-Doi media and using the hot wat /shaker apparatus typically occurred over a duration. | recalled occasional instances where, after starting the procedure on Friday, returned on Monday to find the Ba vegetative cells had not exhausted their Leighton-Doi media nutrients and therefore had not under gone sporulation, or were in various stages of sporulation (i.e. a mix of_vegetative cells, cells under going sporulation, and spores) [~ Jaleo noted on other occasions vegetative cell growth was too rampant. In Situations like this, the Ba spores that were formed were of poor quality and "clumpy". advised the ratio of Ba vegetative cells to media volume was directly proportional to spore quality: too much Ba vegetative cells within a defined media volume results in poor Ba spore quality. [__Jadvised it was standard procedure to allow the Ba vegetative cells to exhaust their nutritive supply in order to induce sporulation. indicated that[~_]research dees not involve dry Ba. The spores were never dried. | Jadvised tol ] knowledge there was no "dry work" D.rather it ed_in "dry work". Typicall pertain only to an experiment the_use of wet_or liquid spore preps. i that was involved in that used infection, and this 1s advantageous in studying certain aspects of