◆ SpookStack

. FD-302a (Rev. 10-6-95) 279A-WF-222936-USAMRIID Bruce E. Ivins. ,On_09/08/2004_,Page Continuation of FD-302 of 10 germination. Ivins first became aware of heat shocking back in the b6 large NHp study. The Division Chief did not want the spores b7F Renograffin purified because that was not was done in the old days. Spores used to be heat shocked at. temperature and refractile spores are not killed. Spores that are used for intramuscular (Im) or aerosol challenges are heat shocked.. All anyone has studied virulence with respect to heat shock. However, did some studies on heat shocked material versus non-heat shocked material. IvINs noted that he told ssA. that USAMRIID used Tween to break up a thick spore suspensions.. who works for has added anti-foam to spore suspensions.. IVINs then discussed b6 b7C the spore harvest procedure. Ames strain Ba, from a primary subculture, is streaked onto a sheep's blood agar (sBA) plate. Three "typical" colonies are selected and placed in phosphate buffer solution, then into Leighton and Doi broth for three days. Spores are harvested and run through a Hypaque density gradient. Hypaque is what is used now, where as Renocal or Renograffin is what was used in the past. Ivins learned this density gradient purification procedure from Ivins advised that he recently received an e-mail from UsAmRIID command that stated the FBI was requesting all files, e-mails, etc. that related to the Ames strain of Ba. IviNS opined that this was a large undertaking and demonstrated this term "Ames" on his archived e-mails, he retrieved over 12o0 e-. mails. Ivins advised that he will be glad to provide whatever. of time. Fifteen laboratory notebooks that were provided by. IviNs were returned to him. An FD-597 was executed and detailed the numbers of the notebooks returned. Ivins was provided with a copy of the FD-597, and the remaining two copies are maintained in an FD-340, in the 1A section of the file.