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The previous chapters have established the following main conclusions: First, the physical evidence associated with the anthrax spores in the attack letters did not reveal the source of the materials in the letters. Second, genetic analyses of the spores established that the anthrax belonged to a particular subtype known as the Ames strain. Third, while no genetic differences were observed between the predominant Bacillus anthracis genotype in the letters and the canonical Ames strain, FBI contractors discovered several subpopulations of B. Anthracis cells in the letters that each produced distinct colony morphologies after growth on agar plates, and some of these subpopulations possessed specific genetic mutations that could be identified by specific sequence-based assays developed by the FBI and its contractors. In this chapter, we describe the establishment of the FBI Repository (FBIR) of Ames strain samples, created by the FBI to look for the B.

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Anthracis subpopulations found in the letters with the hope of being able to identify the source of the B. Anthracis spores used in the attacks. The chapter also outlines the history of RMR-1029, the spore-containing flask at the U.S. Army Medical Research Institute for Infectious Diseases (USAMRIID) that became a focus of the FBI investigation, at least in part because it contained all of the subpopulation genotypes that were assayed.

The results obtained from screening the repository samples are described, including the evidence that appeared to implicate RMR-1029 as the source of the spores in the attack letters. Particular attention is given to the limitations of applying formal statistical methods to these results and of trying to quantify the strength of the relationship between the spores in the attack letters and those in RMR-1029. The committee then discusses evidence bearing on a disputed sample submitted by the suspect in this case as well as follow-up experiments performed by the FBI to determine whether that sample had come from its stated source, RMR-1029 (see Amerithrax Investigative Summary. USDOJ, 2010, pp. 75-79, for a description of the circumstances surrounding the disputed sample). The chapter ends with the committee’s major findings regarding the genetic evidence relevant to the source of the B. Anthracis spores used in the letter attacks.

At the time of the anthrax mailings, the federal government had not systematically collected information on which laboratories possessed anthrax. Although the “Select Agents” program was created in 1996 by the Antiterrorism and Effective Death Penalty Act (Public Law 104-132), this statute governed primarily the transfer of biological agents between research laboratories.

The Act directed the Secretary of Health and Human Services (HHS) to issue regulations governing the transport of biological agents with the potential to pose severe threats to public health and safety through their use in bioterrorism, the so-called “select agents” (NRC, 2009). The authority to regulate select agents was delegated by the HHS Secretary to the Centers for Disease Control and Prevention (CDC). To ensure that these agents were transferred only between responsible parties, CDC required that any laboratories that might transfer select agents be registered and that transfers be reported to CDC and conducted under a permitting system. As long as the select agents were not transferred, specific information about the facilities that possessed these agents did not have to be reported (NRC, 2009b). The determination that the Ames strain of B. Anthracis was used in the attacks led to a process by which the FBI searched for and acquired samples of known and accessible derivative stocks of that strain for comparison. As noted in, the Ames strain had been widely distributed among laboratories around the world for research and vaccine trials, so the FBI first had to identify all laboratories that maintained stocks of the strain.

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Next, the Bureau had to obtain samples of these Ames strain derivatives, which would constitute the FBIR and be screened for the presence of the mutant genotypes found in the letters (see ). To this end, the FBI prepared and issued a subpoena that included a protocol for the collection and submission of Ames strain samples (). This subpoena was sent in February 2002 to 16 laboratories or facilities in the United States that had been identified as possessing stocks of the Ames strain. (It was subsequently determined that one of these domestic laboratories did not possess the Ames strain.) In addition to the subpoenas, __________ According to the FBI (discussion with committee, December 11, 2009), 16 U.S. Laboratories were originally identified as candidates for having the Ames strain. According to the DOJ case closing summary, however, only 15 domestic laboratories were confirmed as repositories of the Ames strain (USDOJ, 2010, p.