Purpose: Provide guidance in the preparation and shipment of Bacillus anthracis Ames strain to the United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland. As per FBI request, submit two culture agar slants of each distinct B. anthracis Ames strain stock in your possession, which differs in source or in other parameters prescribed by the requesting Agency.
Tryptic Soy Agar (TSA) Slants (Remel catalog # 08932, or equivalent)
Sterile inoculating loops
Adhesive Labels, permanent, waterproof
STP-100 Infectious Substance Shipping Container (Saf-T-Pac), or equivalent
1. Collect each B. anthracis Ames strain stock as per your institutional inventory and personal knowledge.
2. Prepare a minimum of two TSA slant tubes per stock by prelabeling with permanent waterproof labels. Include the following information on the label: “B. anthracis Ames strain.” Other designators used by your laboratory, date, and your lab name. Additional information for each stock shall be provided separately.
3. A representative sample of each stock shall be used for inoculation of the TSA slants. If the stock is an agar culture, do not use a single colony, but rather use an inoculum taken across multiple colonies. Thawed frozen stocks or other liquid suspensions shall be well mixed prior to transfer of inoculum to the TSA.
4. Inoculate each TSA slants in a zig zag manner over the surface of the agar.
5. Incubate slants at 35-37C for 12-18 hours to confirm culture growth.
6. Individually wrap the slants in packing materials approved for the transport of infectious select agents in accordance with regulations for the shipment of such materials.
Source: FBI Documents, Preparing and Shipping TSA Slants for B. Anthracis Ames.
consent searches were conducted at USAMRIID in Maryland and at Dugway Proving Ground (DPG) in Utah, and a search warrant was executed at a private company in Ohio (Battelle Memorial Institute) in 2004 to ensure that samples were obtained from every stock derived from the Ames strain in those three facilities. Voluntary submissions were also requested from the three foreign
that were subpoenaed from over 100 Biosafety Level 3 (BSL-3) laboratories in the United States. “These records were supplemented by information culled from FBI interviews of scientists working at each of these labs, and through FBI reviews of relevant scientific publications mentioning the Ames strain” (USDOJ, 2010, p. 17).