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», FD-302d (Rev. 10-6-95) 279A-WF-222936-USAMRIID Continuation of FD-302 of Bruce E. Ivins ,On 09/08/2004 , Page needed primarily for aerosol challenges, which is what everyone is currently moving towards. The "1985" stock was used for guinea pig IM challenges around 1990 during the Gulf War, but it was not used for any rabbit challenges at that time. The "1985" stock was also used for some non-human primate (NHP) challenges. In 1993, this stock was used for aerosol challenges. Notebook # has information on those NHP studies. IVINS does not recall if he was making large batches of spores in the late 1980s. Vollum 1B was the strain originally used for batch growth, and later the Ames strain was used. He made spores for a guinea pig aerosol challenge study in the late 1980s, and this study has been published. [In the early 1990s, IVINS made additional spores for a guinea pig aerosol challenge. The large batches of spores are only required for aerosol challenges. In 1995 or 1996, [ not know what In addition, IVINS knew that Perry Mikesell maintained the Ames strain, but he did not know where Mikesell obtained his original stock. IVINS explained that the following persons have grown spores in his lab: prior to 1988, only IVINS was growing spores; from 1988 to 1997 were growing spores; from 1997 to 1999, were growing spores; from 1999 to ’ spores; from 2001 to 1 growing spores; and since 2002, growing spores. IVINS , In 1989, IVINS created a second seed stock from the "1981" original slant, referred _to as the "1989" sample. IVINS reviewed page 17 of Notebook [__] which is dated December 20, 1990. This page discusses the creation of the "1989" sample using the "1981" original slant. IVINS made 100 milliliters (mL) of the "1989" sample at a concentration of 2 x 10° spores per mL. These spores were not run through a density gradient. IVINS wanted to use a density gradient, but the "old timers" at USAMRIID were against it. IVINS cannot recall if he made this second seed stock because the "1985" sample was depleted. He was also unsure if there was any "1989" sample currently remaining at USAMRIID. In 1989, IVINS was working with[ —___] Ce They were looking at the quality o spores grown in Leighton and Doi media versus the quality of spores grown on agar. They found that spores grown on Leighton and Doi media were "hotter", cleaner, nicer, smaller, less 2 b7c