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Continuation of FD-302 of , 0) FD-302a (Rev. 10-6-95) 279A-WF-222936 SUB 302 Bruce Ivins , 08/13/2003 Page 2 £-pERRY MIKBSELL, dated July, 1991, and labeled pelseveS"That the MIKESELL sample was derive stock culture around 1985. The sample labeled and submitted to the Repository in April, 2002 may be from MIKESELL's collection and not directly from ‘stocks of BA. [___————_—sd| submitted IVINS' samples to the Repository and would be able to provide information on the origin sample in the Repositor IVINS began work at USAMRIID in the early 1980s. When IVINS arrived, the scientists who worked on BA cultured BA, or "sub- cultured" BA by taking an inoculation loop, scraping the loop across a plate of multiple BA colonies, and then using the scraped loop to inoculate a fresh culture for BA growth. IVINS was surprised at this method of inoculation. Before beginning work at USAMRIID, IVINS sub-cultured bacteria by using an inoculation loop to "pick" a single colony of bacteria, instead of taking multiple bacterial colonies as done at USAMRIID. The single colony method was used by many scientists so that sub-culturing would not lead to genetic diversity in growing cultures of bacteria. The multiple ~culturing method used at USAMRIID was recommended by a respected scientist in the BA field. In 1983 and e two plasmids present in BA were discovered, and IVINS everted to the single colony method of subculturing. Further, when IVINS reverted to the single colony method, IVINS also used the original 1981 agar slant of the Ames BA and started a fresh culture of Ames to use in his, IVINS, research. IVINS noted that MIKESELL moved to Ohio several years ago and no longer worked at USAMRIID. MIKESELL was considered an alcoholic while MIKESELL worked at USAMRIID, due to MIKESELL drinking a lot ~of hard liquor. MIKESELL died last year and IVINS attributed MIKESELIYY death to excessive drinking, lack of exercise, and a poor Individuals who worked with MIKESELL at USAMRIID were, ame unknown (LNU), (phonetic) . ro ren