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To: Washington m2 From: Washington Field Re: 279A-WF-222936-USAMRIID, 1/11/2006 PREPARATION OF AMES CHALLENGE MATERIAL: The Ba used in the challenges was i j j j [| of Building[L___] by Ivins. According to bé prior to a challenge, Ivins transported a large ask containing Se: the liquid preparation of _Ba_ to be used for the challenges. The b2 flask was stored in room From the flask, the investigators bIF prepared 15 ml conical tubes filled with 10 mi of Ba solution. The tubes were kept in the refrigerator in room[__]of Building until needed for the challenge. Information provided during other interviews contradicts the information provided by and states that Ivins would prepare the conical tubes in su and transport only the tubes, and not the entire flask, to Building[ | This portion of the investigation remains ongoing. on the first floor of Building but post-challenge material was stored there. The length ime left-over Ames was stored b2 remains controversial. One unresolved contradiction is that some bIF interviewees said that if a conical tube was missing from room it would be noticed since they knew how many tubes they prepared. Other interviewees said that extra tubes were prepared in case one was dropped or damaged, so there were more tubes than test animals. The left-over material was not carefully inventoried or tracked. No one would notice if liquid Ames was taken out of the tubes and replaced with water. No pre-challenge “af kept in the walk-in cooler ‘e) The spores were heat-shocked prior to the challenge to remove vegetative matter and stimulate germination. The Ba Ames spores were suspended in a water solution. The starting concentration of that suspension, which was transferred to the collison nebulizer, was normally 10° colony forming units (cfu) per ml. The AGIs captured samples of the air inside the exposure