Guidance on the Inventory of Select Agents and Toxins: Long-term Storage Criteria for Select Agents

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Section 17(a)(1) of the select agent regulations explains that long-term storage is the placement of select agents or toxins in a system designed to ensure viability for future use, such as in a freezer or other storage container or lyophilized materials. In addition to freezers and other storage containers and lyophilized materials, the system selected by the entity to preserve specimens for future use can be appropriate to specific select agents, and therefore can include, but is not limited to, refrigerators, liquid nitrogen tanks, and room temperature storage units. As a general rule, long-term storage materials include select agents (including viral genetic elements, recombinant and/or synthetic nucleic acids, and organisms containing recombinant and/or synthetic nucleic acids) and nucleic acids encoding functional forms of select toxins and infectious forms of select agent viruses, which are not part of an ongoing experiment or have not been accessed for a significant period of time (e.g., within 30 calendar days).

All select agent materials stored long-term must be kept in a secure location registered with FSAP. All personnel with access to select agent materials must have security risk assessment (SRA) approval and be in compliance with the select agent regulations. In addition, personnel that access Tier 1 select agents must meet additional pre-access suitably and ongoing assessment requirements (Please see the Guidance for Suitability Assessment for more details). Sections 11(f), 14(e) and 15(b) of the select agent regulations outline all of the requirements for possessing Tier 1 agents.

Indicators of long-term storage materials

  1. The material is in a highly concentrated state and would not be used in its present state without dilution to a less concentrated state.
    1. Example: A vial containing a high concentration of select agent bacteria is removed from storage and used to inoculate several tubes of broth, and then the vial is returned to storage.
    2. Example: A vial containing a high concentration of select agent bacteria is removed from storage to make additional aliquots (vials) of highly concentrated bacteria from the original stock(s).
    3. Example: Subcultures of highly concentrated select agent bacteria or high titer select agent viruses (plates, broth cultures, cell culture tubes, flasks, etc.) are used to replace the original seed stocks for experiments to be performed within a defined period of time (e.g., within 30 calendar days).
  2. The material is not part of an ongoing experiment and will not be used for any work by the entity within a defined period of time (e.g., within 30 calendar days).
    1. Example: A vial of select agent is not planned for use for any entity research project, diagnostic procedure, quality control or other laboratory activity within the defined period of time.
  3. The material is not consumed within a defined period of time by the entity (e.g., within 30 calendar days).
    1. Example: A vial of select agents is received by the laboratory but there are no plans to use the contents of the vial for any work within the defined period of time.
    2. Example: A select agent aliquot is collected from an experimental protocol that is preserved for future analysis within the defined time period.
  4. The material is placed in an environment where there is infrequent access to the environment.
    1. Example: Viruses are placed in a liquid nitrogen tank that is only accessed infrequently by a member of the laboratory (e.g., within 30 or more calendar days).

Working stock criteria for select agents

Select agents are considered working stock if the materials are part of an ongoing experiment, accessed frequently, or are not stored for an extended period of time. The select agent regulations do not require entities to maintain inventory records in accordance with Section 17(a)(1) for working stock. Entities should be able to clearly identify how and why material is classified as working stock.

All working stock select agent materials must be kept and used in a secure location registered with FSAP. All personnel with access to working stock select agent materials must have a current SRA approval and be in compliance with the select agent regulations.

Indicators of working stock materials

  1. The material has been diluted from a concentrated state and placed into multiple aliquots in the less concentrated form for immediate use (e.g., within 30 calendar days or as needed by the ongoing experiment).
  2. The material is part of an ongoing experiment and will be used for work by the entity within a period of time as defined by experimental
    1. Example: The material (bacteria, virus) has been propagated for a specific experiment and will be used to infect animals or
    2. Example: A plant infected with a slow growing fungus is maintained at room temperature for a defined period of time (e.g., within 30 calendar days or as dictated by the ongoing experiment).
  3. The material is consumed as part of an ongoing experiment within a defined period of time by the entity (e.g., within 30 calendar days or as dictated by the ongoing experiments).
  4. The material is placed in an environment where there is frequent access to the material, such as a refrigerator or incubator in an active laboratory to support ongoing experiments.

Select agent inventory records

Select agent material includes confirmed clinical specimens, laboratory cultures containing select agents and recombinant and/or synthetic select agent organisms, as well as genetic elements and recombinant and/or synthetic nucleic acids encoding regulated genetic material. This also includes animals, animal tissues, plants, and plant tissues intentionally or accidentally exposed to or infected with a select agent. Each sample of select agent material stored long-term should correspond to a written record containing all of the information required in Section 17(a)(1-2) of the select agent regulations. Final disposition of the material is required to be recorded (e.g., material is no longer in the possession of the entity due to consumption, transfer, destruction). An entity is responsible for maintaining an accurate and current inventory of all toxins possessed regardless of the amount in their possession as stated in section 7(b). All select agents held in long-term storage, may be subject to verification during the inspection. Please review the Security Guidance document for information regarding inventory reconciliations.

Select toxins and select toxin-exposed animals and animal tissues

Entities must maintain records containing all of the information required in Section 17(a)(3). Note that for toxin, there is no differentiation between working stock and long-term storage, because all regulated toxin material that the entity is registered for must be entered into the inventory records. The current quantity of each vial must be documented for toxins following each use. The current quantity of each vial that is recorded following the last usage may be examined during an inspection.

Individuals may not access BSAT unless approved by FSAP following a SRA. Entities that ship aggregate amounts of select toxin below the amounts listed in 42 CFR § 73.3(d)(3) are not required to submit an APHIS/CDC Form 2 for approval prior to shipment; however, the entity must use due diligence and maintain documentation of the amount of shipped toxin in order to fulfill their responsibilities under Section 73.16(l). Please see the Select Toxin Guidance for more information.

Once an animal has been injected with or exposed to a select toxin (for example, by inhalation, dermal absorption, or ingestion), the animal is not considered a “select toxin” and does not need to be housed in a registered space (See 42 CFR § 73.3(d)(3)).   Please note that accurate and current accounting are required for any animals or plants intentionally or accidentally exposed to or infected with a select agent (including number and species, location, and appropriate disposition) (See 42 CFR § 73.17(a)(2).

Until the select toxin is injected into or exposed to the animal, the select toxin is subject to the select agent regulations. This includes storage or use of the material (e.g., injection or exposure procedure). If the select toxin is stored or used in the same area as the injected or exposed animal, that area will need to be listed on an entity’s approved certificate of registration.

The room where select toxin injection or exposure procedures occur may be assessed using laboratory biosafety level criteria instead of animal laboratory biosafety level criteria. These rooms, however, must be included on an entity’s registration and must be managed as regulated space.

Select agent-infected animals and animal tissues

Section 17(a)(2) of the select agent regulations requires an accounting of animals intentionally or accidentally exposed to or infected with a select agent (including number and species, location, and appropriate disposition). The intent of this requirement is to ensure that an entity has a system in place to manage and account for the number of research animals used in experimental protocols with select agents. The accounting should include the location of these animals in the facility and their final disposition. This information can be contained in research notes, animal care logbooks, daily check sheets, or other formats. An accidental release of exposed animals or animal exposure to select agents outside of primary containment must be reported using the APHIS/CDC Form 3 (Theft, Loss and Release form), which requires an entity to provide a detailed description and follow-up actions of the incident. An accounting of animals intentionally or accidentally exposed to or infected with a select agent (including number and species, location, and appropriate disposition) will facilitate the investigation of these types of incidents.

Fluid, serum, tissue, or other samples collected from animals infected with or exposed to select agents in the laboratory are considered select agent material, and are subject to the select agent regulations, under Section 17(a)(1). Animals that are infected with select agents for experiments that require longer incubation periods (e.g., within 30 days or as mandated by the experimental protocol) before the agent or disease can be detected are considered to be select agents and must be handled in the same manner as infected animals above. A uniquely identified vial-by-vial inventory record of tissue specimens from a single source or experiment is not required for animals or animal samples, and specimens may be grouped without individual vial identification.

For example, 14 vials of infected mouse lung samples from the same experiment may be grouped under one reference identification number, consistent with the experimental protocol. However, the reference number must be linked to a record that notes 14 vials upon inventory reconciliation. If an entity wishes to group specimen vials in this manner, each primary and secondary container in which the samples are stored should be labeled with the date placed in storage, the agent contained in the sample, and a reference identification that is associated with a written record (inventory record, research notes, etc.) describing those specimens, including the total number of vials. The entity is required to maintain an inventory of all specimens, therefore, an entity using this method would need to have an accurate accounting for each identification number. This should include all of the information required in Section 17 and an entity must be able to account for each vial associated with that number to ensure compliance. As vials are removed from inventory for processing, their removal must be noted in the written record to maintain an accurate and current inventory.

Samples collected from animals that are presumed to have been naturally infected (i.e., not intentionally introduced) would not be considered select agent material and are not required to be handled as regulated material until the samples have been confirmed to contain select agent material. Diagnostic samples from which the presence of a select agent has been detected are subject to the select agent regulations; however, diagnostic samples from which only an antibody response to a select agent is detected are not considered select agent material. In addition, samples taken from animals experimentally infected with select agents are considered select agents and are subject to the select agent regulations, unless the absence of the agent can be demonstrated.

Long-term storage criteria for select agent-infected arthropods

Infected arthropods may be recorded individually or pooled, at the discretion of the entity. The long-term storage inventory of infected arthropod vials may be verified during an inspection. Each vial in long-term storage must have an associated record that collects all of the information required in Section 17(a)(1) (See Appendix I).

Long-term storage criteria for select agent-infected plants

The regulations in Section 17(a)(2) require an accurate, current inventory of plants intentionally or accidentally inoculated with a select agent (including number and plant species, location, and appropriate disposition). The intent of this requirement is that an entity has a system in place to manage and account for the number of research plants used in experimental protocols with select agents. The accounting should include the location of these plants in the facility and their final disposition. This information can be contained in research notes, plant logbooks, daily check sheets, or other formats. In the event of a theft, loss, or accidental release outside of primary containment, the incident must be immediately reported to FSAP with a follow-up submission within 7 days on APHIS/CDC Form 3 – Theft, Loss and Release. The submission of the APHIS/CDC Form 3 will provide a detailed description and follow-up actions of the incident.

Plants that are infected with select agents for experimental purposes or as a mechanism to maintain long-term viability and are held for long incubation periods (e.g., within 30 days) are considered long-term storage select agents and require accurate inventory records for each inoculated plant, such as an identification number that includes all of the information required in Section 17(a)(1) and should be handled as select agent material. Plants infected with select agents for experiments that require longer incubation periods (e.g., within 30 calendar days or as mandated by the experimental protocol) before the agent can be detected or disease symptoms detected are considered long-term storage select agents and must be handled in the same manner as infected plants above. These infected plants can be removed from long-term storage inventory records when they are shown to be uninfected by appropriate testing (documentation must be made available upon request).

Select agent infected plant tissues (living or dead) that contain infectious select agent propagules are considered long-term storage, as are culture plates and vials containing isolated select agents and are subject to regulation and should correspond to a written record that includes all of the information required in Section 17(a)(1). If several vials or culture plates generated from a single source of material are in long-term storage, they may be grouped under a single reference identification number; the inventory records must indicate how many vials are associated with this reference identification number and a written record must be maintained of the vials and plates removed for accurate inventory reconciliation. The entity must be able to account for each vial or plate associated with the reference identification number. Plants inoculated from a single source and held in long-term storage will be considered individually.  Each must be given a unique reference identification number for inventory records and must correspond to a written record that includes all of the information required in Section 17(a)(1) (). As vials or plates are removed from inventory for processing, their removal must be noted in the written record to maintain an accurate and current inventory.