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~ FD-302a (Rev. 10-6-95) 279A-WF-222936-USAMRIID Continuation of FD-302 of Bruce E. Ivins. On 09/08/2004 Page 2 needed primarily for aerosol challenges, which is what everyone is currently moving towards. The "1985" stock was used for guinea pig IM challenges around 199o during the Gulf war, but it was not used for any rabbit challenges at that time. The "1985" b6 stock was also used for some non-human primate (Nhp) challenges. b7C 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 Imade 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, lIvInS does not know what. In addition, Ivins knew that Perry Mikesell maintained the Ames strain, but he did not know where Mikesell obtained his original spores in his lab: prior to 1988, only Ivins was growing spores; from 1988 to 1997, IVINS were growing spores; from 1997 to 1999 were growing spores; from 1999 to 2001, were growing spores; from 2001 to 2002, were growing spores; and since 2002, Ivins have been growing spores. 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 b6 b7C using the "198i" original slant. Ivins made 100 milliliters (mL) of the "1989" sample at a concentration of 2 x 109 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 Agaunnane andees 68nt Aue sem aanya dt aanaun oare sem remaining at USAMRIID. In 1989, IviNs was working with. They were looking at the quality of. 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