Federal Select Agent Program - General FAQ's about Select Agents and Toxins

General FAQ's about Select
Agents and Toxins

FAQ's by Topic

General

If I have questions regarding the Federal Select Agent Program or select agents and toxins, where can I go to get help?

For specific questions that aren't addressed at http://www.selectagents.gov/, please submit an email directly to the Federal Select Agent Program at:
CDC: LRSAT@cdc.gov
APHIS: AgSAS@aphis.usda.gov
What are the select agent regulations?

The select agent regulations can be found at 42 CFR Part 73, 9 CFR Part 121, and 7 CFR Part 331. The select agent regulations implement the Subtitle A and Subtitle B (also known as the Agricultural Bioterrorism Protection Act of 2002) of the Public Health Security and Bioterrorism Preparedness and Response Act of 2002, setting forth the requirements for the possession, use, and transfer of select agents and toxins.
What is the Federal Select Agent Program?

The Centers for Disease Control and Preventions' (CDC) Division of Select Agents and Toxins (DSAT) and the Animal and Plant Health Inspection Services' (APHIS) Agriculture Select Agent Services (AgSAS) jointly constitute the Federal Select Agent Program.
What is an entity?

An entity is any government agency (Federal, State, or local) academic institution, corporation, company, partnership, society, association, firm, sole proprietorship, or other legal entity. See section 1 of the select agent regulations.
What is a select agent or toxin?

Pursuant to 42 USC 262a and 7 USC 8401, select agents and toxins are a subset of biological agents and toxins that the Departments of Health and Human Services (HHS) and Agriculture (USDA) have determined to have the potential to pose a severe threat to public health and safety, to animal or plant health, or to animal or plant products. The current list of select agents and toxins can be found at 42 CFR §§ 73.3, 73.4, 9 CFR §§ 121.3, 121.4, and 7 CFR § 331.3.

A current list of select agents and toxins can also be found at: http://www.selectagents.gov/SelectAgentsandToxinsList.html.

What is a Tier 1 Select Agent or Toxin?

A subset of select agents and toxins have been designated as Tier 1 because these biological agents and toxins present the greatest risk of deliberate misuse with significant potential for mass casualties or devastating effect to the economy, critical infrastructure, or public confidence, and pose a severe threat to public health and safety:

Tier 1 Select Agents and Toxins
HHS Agents and Toxins	Overlap Agents	USDA Agents
Bacillus cereus Biovar anthracis Botulinum neurotoxins Botulinum neurotoxin producing species of Clostridium Ebola virus Francisella tularensis Marburg virus Variola major virus (Smallpox virus) Variola minor virus (Alastrim) Yersinia pestis	Bacillus anthracis Burkholderia mallei Burkholderia pseudomallei	Foot-And-Mouth Disease virus Rinderpest virus

Entities that possess, use, or transfer Tier 1 select agents and toxins must adhere to the additional requirements detailed within the Select Agent Regulations.

Does my entity have to meet Tier 1 requirements if we only store a Tier 1 agent or toxin?

Yes, the requirements apply to both possession and use of Tier 1 select agents or toxin.
How is the select agent list determined?
In determining whether to include an agent or toxin on the HHS select agent list, the "Public Health Security and Bioterrorism Preparedness and Response Act" requires that the following criteria be considered:
- The effect on human health of exposure to the agent or toxin;
- The degree of contagiousness of the agent or toxin and the methods by which the agent or toxin is transferred to humans;
- The availability and effectiveness of pharmacotherapies and immunizations to treat and prevent any illness resulting from infection by the agent or toxin; and
- Any other criteria, including the needs of children and other vulnerable populations, that the Secretary considers appropriate.
In determining whether to include an agent or toxin on the USDA list, the "Agricultural Bioterrorism Protection Act of 2002" requires that the following criteria be considered:
- The effect of exposure to the agent or the toxin on animal and plant health, and on the production and marketability of animal or plant products;
- The pathogenicity of the agent or the toxin and the methods by which the agent or toxin is transferred to animals or plants;
- The availability and effectiveness of pharmacotherapies and prophylaxis to treat and prevent any illness or disease caused by the agent or toxin; and
- Any other criteria that the Secretary considers appropriate to protect animal or plant health, or animal or plant products.
The lists are required to be reviewed and republished every 2 years, or revised as necessary.
Why do I need to register with the Federal Select Agent Program for select agents or toxins that I possess, use, or transfer?

Registration of individuals or entities possessing, using, or transferring these select agents and toxins, unless the agent or toxin is specifically excluded by the select agent regulations or the individual or entity has been specifically exempted by the Federal Select Agent Program, is required pursuant to the select agent regulations. Anyone violating the provisions of 42 CFR Part 73, 7 CFR Part 331, or 9 CFR Part 121 may be subject to civil and/or criminal penalties including imprisonment for not more than 5 years.
Where can I obtain any forms to submit requests or reports associated with the select agent regulations?

Forms are available at http://www.selectagents.gov/forms.html.
What are the penalties for not being in compliance with the regulations?
- Administrative:
- Civil:
- Criminal: Violations of 18 USC 175b
What is the Corrective Action Plan program?

In March 2008, the Federal Select Agent Program established the Corrective Action Plan (CAP) program to assist entities, identified by Federal Select Agent Program as having systemic biosafety and security deficiencies, to come into compliance with the select agent regulations. After being given the opportunity to participate, an entity begins by submitting a detailed plan, including target completion dates, addressing the specifics of how the entity will address and correct regulatory deficiencies identified by Federal Select Agent Program. An entity's participation in a CAP allows Federal Select Agent Program to both provide technical assistance as well as monitor an entity's progress in correcting security or safety shortcomings.
Under what conditions could a certificate of registration be withdrawn, revoked, or suspended?
The Federal Select Agent Program will withdraw a certificate of registration upon the written request of the entity , if the entity no longer possesses or uses any select agents or toxins and no longer wishes to be registered.

A certificate of registration may be revoked or suspended if:
- The individual or entity, the Responsible Official, or an individual who owns or controls the entity is a restricted person (within any of the categories described in 18 U.S.C 175b).
- The individual or entity, the Responsible Official, or an individual who owns or controls the entity is reasonably suspected by any Federal law enforcement or intelligence agency of:
- The individual or entity does not meet the requirements of select agent regulations.
- It is determined that such action is necessary to protect agricultural or public health and safety.
Is there a restriction on the publication of select agent information?

The select agent regulations place no specific restrictions on the publication of research related to select agents or toxins. However, any records or information systems that could allow an individual to gain access to the select agents or toxins should be safeguarded to prevent unauthorized access, theft, loss, or release of these materials. The Federal Select Agent Program strongly encourages entities to refrain from publishing detailed information about locations of select agents and toxins, quantities on site, or researchers.
Should the plans (i.e., biosafety, incident response, and security) developed to comply with the select agent regulations be available to only those individuals that have access approvals from the Federal Select Agent Program?

The select agent regulations place no specific restrictions on the biosafety, incident response, or security plan being made publically available to individuals (e.g., first responders, maintenance workers) that don't have access approvals from the Federal Select Agent Program. However, any records or information systems that could allow an individual to gain access to the select agents or toxins should be safeguarded to prevent unauthorized access, theft, loss, or release of these materials. The Federal Select Agent Program strongly encourages entities to refrain from publishing detailed information about locations or quantities of select agents and toxins on site.
How can I report suspicious activity that may be criminal in nature concerning a registered entity, its personnel, or its select agents or toxins?

You can notify a Federal Bureau of Investigation Weapons of Mass Destruction Coordinator of any suspicious activity that you believe may be criminal in nature concerning an entity, its personnel, or its select agents or toxins. To locate your Federal Bureau of Investigation Weapons of Mass Destruction Coordinator, refer to http://www.fbi.gov/contact-us/field.
Is it possible for me to report a safety or security issue related to a select agent or toxin without identifying myself?

The Federal Select Agent Program has established a confidential means for reporting safety and security issues associated with the possession, use and transfer of select agents and toxins. The Department of Health & Human Services, Office of Inspector General (OIG) maintains a hotline that allows individuals to anonymously report fraud, waste and abuse in all departmental programs. This hotline is now available to anonymously report safety or security issues related to select agents and toxins. When reporting these issues to OIG, please ensure that you indicate it is a "Select Agent Program" issue.

OIG Hotline contact information:

Voice:
1-800-HHS-TIPS (800-447-8477)

Fax:
1-800-223-8164

Web:
https://oig.hhs.gov/fraud/report-fraud/index.asp

Mail:
Office of Inspector General
Department of Health & Human Services
Attn: Hotline
P.O. Box 23489
Washington, DC 20026
What are the requirements for entities that are registered to possess Tier 1 select agents and toxins?
The requirements specifically for Tier 1 select agents and toxins are listed below. These requirements are contained in 42 CFR Part 73 and 9 CFR Part 121:
1. Sections 3 and 4 (Health and Human Services or United States Department of Agriculture and Overlap Select Agents and Toxins), include the names of the agents and toxins on the Tier 1 list.
2. Sections 5 and 6 (Exemptions for select agents and toxins) require immediate notification for Tier 1 select agents and toxins contained in a specimen presented for diagnosis or verification.
3. Section 11(f) (Security) requires that the security plan must:
  1. Describe procedures for conducting a pre-access suitability assessment of persons who will have access to a Tier 1 select agent or toxin and describe procedures for the ongoing assessment. The on-going assessment procedures must include:
    1. Self- and peer-reporting of incidents or conditions that could affect an individual's ability to safely have access to or work with select agents and toxins, or to safeguard select agents and toxins from theft, loss, or release;
    2. The training of employees with access to Tier 1 select agents and toxins; and
    3. The ongoing suitability monitoring of individuals.
  2. Describe procedures for how an entity's Responsible Official will coordinate their efforts with the entity's safety and security professionals and share relevant information.
4. Section 11(f) (Security) requires that the following security enhancements be developed:
  1. Procedures that will limit access to only approved individuals that are enrolled in entity's suitability assessment program;
  2. Procedures that limit access to laboratory and storage facilities outside of normal business hours to only those specifically approved by the Responsible Official or designee;
  3. Procedures for allowing visitors, their property, and vehicles at the entry and exit points to the registered space, or at other designated points of entry to the building, facility, or compound that are based on the entity's site-specific risk assessment;
  4. A minimum of three security barriers where each security barrier adds to the delay in reaching secured areas where select agents and toxins are used or stored;
  5. All registered space or areas that reasonably afford access to the registered space must be protected by an intrusion detection system (IDS) unless physically occupied and personnel monitoring the IDS must be capable of evaluating and interpreting the alarm and alerting the designated security response force or law enforcement;
  6. For powered access control systems, describe procedures to ensure that security is maintained in the event of the failure of access control systems due to power disruption affecting registered space; and
  7. The entity must determine that the response time for security forces or local police will not exceed 15 minutes or provide security barriers that are sufficient to delay unauthorized access until the response force arrives.
5. Section 12 (Biosafety) requires that the biosafety plan must include an occupational health program for individuals with access to Tier 1 select agents and toxins and those individuals must be enrolled in the occupational health program.
6. Section 14 (Incident response) requires that the incident response plan must:
  1. Describe the entity's response procedures for failure of intrusion detection or alarm system; and
  2. Describe procedures for how the entity will notify the appropriate Federal, State, or local law enforcement agencies of suspicious activity that may be criminal in nature and related to the entity, its personnel, or its select agents or toxins.
7. Section 15 (Training) requires that training must include annual insider threat awareness briefings on how to identify and report suspicious behaviors.
Note: There are the following additional requirements for security and training for any entity registered to possess select agents and toxins:
1. The security plan must:
  1. Describe procedures for how the Responsible Official will be informed of suspicious activity related to the entity, its personnel, or its select agents or toxins that may be criminal in nature and for how the entity will notify the appropriate federal, state, or local law enforcement agencies of such activity.
  2. Contain provisions for information security. The provisions must:
    1. Ensure that all external connections to systems which manage security for the registered space are isolated or have controls that permit only authorized and authenticated users;
    2. Ensure that authorized and authenticated users are only granted access to select agent and toxin related information, files, equipment (e.g., servers or mass storage devices) and applications as necessary to fulfill their roles and responsibilities, and that access is modified when the user's roles and responsibilities change or when their access to select agents and toxins is suspended or revoked;
    3. Ensure that controls are in place that are designed to prevent malicious code (such as, but not limited to, computer virus, worms, spyware) from compromising the confidentiality, integrity, or availability of information systems which manage access to registered spaces;
    4. Establish a robust configuration management practice for information systems to include regular patching and updates made to operating systems and individual applications; and
    5. Establish procedures that provide backup security measures in the event that access control systems, surveillance devices, and/or systems that manage the record-keeping requirements found in Section 17 are rendered inoperable.
  3. Contain provisions and policies for shipping, receiving, and storage of select agents and toxins, including documented procedures for receiving, monitoring, and shipping of all select agents and toxins. These provisions must provide that an entity will properly secure containers on site and have a written contingency plan for unexpected shipments.
2. Security enhancements state that entities must conduct complete inventory audits of all affected select agents and toxins in long-term storage when any of the following occur:
  1. Upon the physical relocation of a collection or inventory of select agents or toxins for those select agents or toxins in the collection or inventory;
  2. Upon the departure or arrival of a principal investigator for those select agents and toxins under the control of that principal investigator; or
  3. In the event of a theft or loss of a select agent or toxin, all select agents and toxins under the control of that principal investigator.
3. Training must include security awareness and incident response.
What is an SA Gram?

A “Select Agent Gram” or “SA Gram” is an email sent from the Federal Select Agent Program (FSAP) to the regulated community (i.e., Responsible Officials and Alternate Responsible Officials) that communicates information from the FSAP.

If anyone besides the Responsible Officials and Alternate Responsible Officials would like to receive FSAP information, FSAP offers the GovDelivery Email Subscription Management System. The System allows website visitors to subscribe to receive email based on their interest in the program. Messages in this system will be of a general nature and will not contain sensitive select agent information. To sign up for updates, the individual would just need to provide his/her e-mail address for the “Subscribe to Federal Select Agent Program email updates” located in the footer at: http://www.selectagents.gov/.
Where do I find FSAP’s Regulatory Interpretations?

To see the web page with FSAP’s Regulatory Interpretations, see http://www.selectagents.gov/regulations.html.
How do I submit a request for regulatory interpretation?

To submit an interpretation request, an individual submits the request to the Federal Select Agent Program. The request should contain the regulatory requirement of concern, issues regarding the application of the regulatory requirement, and where appropriate, the requestor’s desired outcome.

CDC: the request can be submitted to DSAT via email (lrsat@cdc.gov), FAX (404-718-2096), or mail (Division of Select Agents and Toxins, 1600 Clifton Rd MS A-46, Atlanta, GA 30329)

or

APHIS: the request can be submitted to APHIS via email (AgSAS@aphis.usda.gov), FAX (301-734-3652) or mail (Animal and Plant Health Inspection Service, Agriculture Select Agent Services, 4700 River Road, Unit 2, Mailstop 22, Cubicle 1A07, Riverdale, MD 20737).

In response to the request, the Federal Select Agent Program will provide a written interpretation. The interpretation will also be posted at: http://www.selectagents.gov/regulations.html.

Select Agents and Toxins

An entity possesses a select toxin in an aggregated amount that exceeds the excluded limit listed in the select agent regulations; but the select toxin is distributed among multiple Principal Investigators, none of whom individually controls an amount above the excluded limit. Does this entity have to register with the Federal Select Agent Program?

No, if the aggregate amount of the select toxin under the control of each Principal Investigator (i.e., individual who is designated by the entity to direct a project or program and who is responsible to the entity for the scientific and technical direction of that project or program) at the same entity, does not, at any time, exceed the select toxin amounts listed in 42 CFR 73.3(d)(7).
How do the select agent regulations apply to a Principal Investigator (PI) possessing a regulated amount of a select toxin who transfers an “unregulated amount” of a select toxin to another person? Note: An “unregulated amount” of a select toxin would be those amounts listed in 42 CFR § 73.3(d)(7).

A PI transferring (meaning the PI will no longer have control of the select toxin) an unregulated amount of a select toxin to another person is not required to obtain prior approval for the transfer using the APHIS/CDC Form 2. The PI will be required to exercise due diligence prior to the transfer and keep a record of the transfer. The PI will be required to note the amount of the transfer on the PI’s select toxin inventory. Please note that transfer of any amount of select toxins outside of the U.S. requires an export license from the Department of Commerce, Bureau of Industry and Security (See 15 CFR Supplement #1 to Part 774, Export Control Classification Number 1C351).

NOTE: If a PI moves any amount of a select toxin to someone the PI supervises at the same entity (i.e., a research assistant), such that the toxin is still under the control of the PI, then that movement is not considered a transfer within the meaning of the regulations. Therefore, the select toxin would not be excluded from the regulation and the individual and room where the toxin is used or stored would need to be listed on the registration and must meet all requirements of the select agent regulations. For example, the PI maintains two laboratories. One laboratory stores above the excluded amount of the toxin. The second laboratory is where the PI’s research assistant works with the excluded amount of toxin. Since the PI maintains control over both portions of the select toxin, both rooms must be listed on the registration and must meet all requirements of the select agent regulations.
What genomic material from select agents are subject to the select agent regulations?
The following genetic elements, recombinant and/or synthetic nucleic acids, and recombinant and/or synthetic organisms are select agents (See section 3(c) of 42 CFR Part 73, 9 CFR Part 121, and 7 CFR Part 331):
- Nucleic acids that can produce infectious forms of any of the select agent viruses.
- Recombinant and/or synthetic nucleic acids that encode for the functional form(s) of select agent toxins if the nucleic acids:
  - Can be expressed in vivo or in vitro or,
  - Are in a vector or recombinant host genome and can be expressed in vivo or in vitro.
- Select agents and toxins that have been genetically modified.
Additional information is available at: http://www.selectagents.gov/SyntheticGenomics.html.
Under what conditions is a select agent or toxin excluded from the select agent regulations?
The Federal Select Agent Program does not regulate a select agent or toxin if it is:
- in its naturally occurring environment, provided that it has not been intentionally introduced, cultivated, collected, or otherwise extracted from its natural source
- non-viable select agents or nontoxic toxins.
- an attenuated strain of a select agent or an inactive form of a select toxin that has been determined by the Federal Select Agent Program not to pose a significant threat to public health or safety, to animal health, or to animal products. The list of exclusions can be found at http://www.selectagents.gov/SelectAgentsandToxinsExclusions.html.
- a select agent or toxin that has been subjected to decontamination or a destruction procedure when intended for waste disposal.
- a select agent or regulated nucleic acids that can produce infectious forms of any select agent virus that has been subjected to a validated inactivation procedure that is confirmed through a viability testing protocol. Surrogate strains that are known to possess equivalent properties with respect to inactivation can be used to validate an inactivation procedure; however, if there are known strain-to-strain variations in the resistance of a select agent to an inactivation procedure, then an inactivation procedure validated on a lesser resistant strain must also be validated on the more resistant strains.
- material containing a select agent that is subjected to a procedure that removes all viable select agent cells, spores, or virus particles if the material is subjected to a viability testing protocol to ensure that the removal method has rendered the material free of all viable select agent.
- a select agent or regulated nucleic acids that can produce infectious forms of any select agent virus not subjected to a validated inactivation procedure or material containing a select agent not subjected to a procedure that removes all viable select agent cells, spores, or virus particles if the material is determined by the HHS Secretary to be effectively inactivated or effectively removed. To apply for a determination an individual or entity must submit a written request and supporting scientific information to CDC. A written decision granting or denying the request will be issued.
- an select toxin identified in an original food sample or clinical sample.
- for those laboratories that are not exempt under § 73.5 (a) and § 73.6 (a), Botulinum neurotoxin that is produced as a byproduct in the study of Botulinum neurotoxin producing species of Clostridium so long as the toxin has not been intentionally cultivated, collected, purified, or otherwise extracted, and the material containing the toxin is rendered non-toxic and disposed of within 30 days of the initiation of the culture.
- waste generated during the delivery of patient care by health care professionals from a patient diagnosed with an illness or condition associated with a select agent, where that waste is decontaminated or transferred for destruction by complying with state and Federal regulations within seven calendar days of the conclusion of patient care.
- toxin in an amount under the control of a principal investigator, treating physician or veterinarian, or commercial manufacturer or distributor does not exceed, at any time, the amounts indicated in the list below:
Do laboratories working with toxigenic strains of Staphylococcus have to register with the Federal Select Agent Program?

Laboratories that work with strains of Staphylococcus (non-select agent) that produce Staphylococcal enterotoxins (subtypes A-E) (select toxin) are not regulated so long as the select toxin isn't intentionally, cultivated, collected, or otherwise extracted from its natural source. If work with toxigenic strains of Staphylococcus result in the production of Staphylococcal enterotoxins (subtypes A-E), whether it was intentional or not, the work would be subject to the regulations if the amount of toxin produced is above the permissible toxin amount of 100 mg of Staphylococcal enterotoxins (subtypes A-E).
How do I apply for an exclusion of an attenuated strain of a select agent or a select toxin modified to be less toxic or potent?
To apply for the exclusion of an attenuated strain of a select agent or a select toxin modified to be less toxic or potent, an applicant must submit the request to the Federal Select Agent Program. Requests for exclusions of a select agent or toxin should be sent to the entity's lead agency (CDC or APHIS).

CDC: the request can be submitted to CDC via email (lrsat@cdc.gov), FAX (404-718-2096), or mail (Division of Select Agents and Toxins, 1600 Clifton Rd MS A-46, Atlanta, GA 30329)

or

APHIS: the request can be submitted to APHIS via email (AgSAS@aphis.usda.gov), FAX (301-734-3652) or mail (Animal and Plant Health Inspection Service, Agriculture Select Agent Program, 4700 River Road, Unit 2, Mailstop 22, Cubicle 1A07, Riverdale, MD 20737).

Information should include at a minimum:
- name of the strain/toxin,
- how strain/toxin was derived,
- how the requester ascertained that agent is avirulent or toxin is modified to be less toxic or potent, and
- all citation or pertinent data to support your request.
In response to the request for exclusion of an attenuated strain or toxin modified to be less toxic or potent, the Federal Select Agent Program will provide a written decision granting or denying the request. Exclusions are effective upon notification to the applicant.

Attenuated strains and toxins modified to be less toxic or potent that have been granted an exclusion from the regulations can be found at http://www.selectagents.gov/SelectAgentsandToxinsExclusions.html . Until such time that exclusions are considered and granted, the attenuated strain of the select agent or select toxin modified to be less toxic or potent is subject to the requirements of select agent regulations.
Do we need to register toxin subunits or fragments?
It depends on the circumstance. If a toxin subunit or fragment is less potent or toxic, then the material is considered excluded from the regulation and therefore not subject to the requirements of 42 CFR Part 73, including the registration and transfer requirements. However, registration is required if toxin subunits or fragments are combined to generate toxic form(s) of any of the toxins and the aggregate amount under the control of a principal investigator, treating physician or veterinarian, or commercial manufacturer or distributor exceeds the amount specified below:
- Abrin 1000 mg
- Botulinum neurotoxins 1 mg
- Short, paralytic alpha conotoxins 100 mg
- Diacetoxyscirpenol (DAS) 10,000 mg
- Ricin 1000 mg
- Saxitoxin 500 mg
- Staphylococcal Enterotoxins (Subtypes A, B, C, D, and E) 100 mg
- T-2 toxin 10,000 mg
- Tetrodotoxin 500 mg
For additional information on nucleic acids that encode for functional forms of a select toxin, please see the Guidance on the regulation of select agent and toxin nucleic acids.
Do we need to register if we are using plasmids to express toxin subunits or fragments?

It depends on the circumstance. If the plasmid does not encode for the functional form of any listed toxin, then the material is considered excluded from the regulation and therefore not subject to the requirements of 42 CFR Part 73, including the registration and transfer requirements. However, if the plasmid encodes for the functional form of a listed toxin (not a fragment of the toxin) and is in a vector or host chromosome and/or can be expressed in vivo or in vitro, then the material would be subject to regulation.
What strains of Clostridium would be characterized as being Botulinum neurotoxin producing species of Clostridium?

Botulinum toxin (serotypes A through G) is produced by Clostridium botulinum (phenotypically classified into 4 Groups -I, II, III, IV) and some isolates of Clostridium baratii (serotype F only), Clostridium butyricum (serotype E only), and Clostridium argentinense (serotype G only). Some strains of C. botulinum have been identified that produce 2 botulinum serotypes (e.g. both A and B).

Demonstration of the production of botulinum neurotoxin, through a toxin detection test, in a suspect culture is critical to the characterization of a Botulinum neurotoxin producing species of Clostridium. Most isolates of C. baratii, C. butyricum, and some isolates of C. argentinense do not contain the genes necessary to produce botulinum toxin. Some non-toxigenic clostridia species (e.g. C. sporogenes) are phenotypically identical to C. botulinum but may result in an identification of C. botulinum on some rapid biochemical identification methods (i.e., 16sRNA sequencing may misidentify a non-toxin producing clostridia as a C. botulinum, particularly Group II strains).

If a laboratory detects botulinum neurotoxin from a culture of any suspected Clostridium species (such as C. botulinum, C. baratii, C. butyricum, and some isolates of C. argentinense), the laboratory will need to report the identification of Botulinum neurotoxin producing species of Clostridium by completing Section 1 of the APHIS/CDC Form 4 within seven calendar days of identification. In addition, if the laboratory is not registered with the Federal Select Agent Program for possession of Botulinum neurotoxin producing species of Clostridium, the laboratory must destroy or transfer the isolate within seven calendar days after identification.
Are chimeric viruses containing genetic material from a select agent considered a select agent?

The FSAP regulates chimeric viruses whose genomes 1) contain the backbone and replication machinery of a select agent virus or 2) contain genes from different select agent viruses. Regulated chimeric viruses are evaluated on a case-by-case basis to determine if the viruses exhibit sufficient attenuation to be excluded. Chimeric viruses that “mix and match” select agent genes with non-select agent genes from the same virus family may be regulated as a select agent virus. It is the entity’s responsibility to determine if the resultant chimera is a select agent; however, the FSAP encourages entities to submit these types of chimeras for review. A list of excluded chimeric viruses is available for the entity to reference.
On September 14, 2016, CDC published an Interim Final Rule adding B. cereus Biovar anthracis to the list of HHS select agents and toxins to be regulated as a Tier 1 agent. Do those working with B. cereus strains need to test each one of their samples to determine if they possess B. cereus Biovar anthracis?

Those currently working with B. cereus strains that have not been identified as B. cereus Biovar anthracis strains do not need to test their samples to determine if they possess B. cereus Biovar anthracis. Because of the agent’s currently limited known distribution, it is unlikely that those currently with samples of B. cereus will have this particular strain of the bacteria unless they have specifically requested it or have previously identified it. Those working with B. cereus would only interact with the Federal Select Agent Program when they want to register for B. cereus Biovar anthracis to obtain it, or if they identify B. cereus Biovar anthracis.

Exclusions – Attenuated Strain

How do I submit a request to exclude from the select agent regulations an attenuated strain of a select agent or a select toxin modified to be less potent or toxic?
To apply for the exclusion of an attenuated strain of a select agent or select toxin that has been modified to be less potent or toxic, an applicant must submit the request to the Federal Select Agent Program. Requests for the exclusion of a select agent or toxin should be sent by the Responsible Official to the registered entity’s file manager. If the applicant is a non-registered entity, please submit the exclusion request to DSAT or AgSAS).

CDC: the request can be submitted to DSAT via email (lrsat@cdc.gov), FAX (404-718-2096), or mail (Division of Select Agents and Toxins, 1600 Clifton Rd MS A-46, Atlanta, GA 30329)
or
APHIS: the request can be submitted to APHIS via email (AgSAS@aphis.usda.gov), FAX (301-734-3652) or mail (Animal and Plant Health Inspection Service, Agriculture Select Agent Services, 4700 River Road, Unit 2, Mailstop 22, Cubicle 1A07, Riverdale, MD 20737).

The exclusion request should contain information that is responsive to these criteria:
- Documented history of not causing disease in humans, or relevant animal or plant models.
- Defined genetic mutations or alterations known to attenuate virulence in humans or relevant animal or plant models.
- Data showing the mutations have a low frequency of reversion to wild-type virulence.
- Level of difficulty in engineering the attenuated strain to restore wild-type virulence. For each pathogen, the sample size and type of animal or plant model used to test virulence is important.
- Quantitative measures demonstrating a change in virulence in an appropriate animal or plant model.
- Information regarding tests that may be conducted to differentiate animals or plants exposed to the attenuated strain from those infected with the wild-type organism.
- Related published scientific papers which support the methods and data provided for the exclusion.
The data requirements necessary for consideration of an exclusion request for highly pathogenic Avian influenza (HPAI) virus are listed on the Federal Select Agent Program website http://www.selectagents.gov/resources/usdatemplate-for-ai.pdf.

In response to the request for exclusion of an attenuated strain or a toxin that has been modified to be less potent or toxic, the Federal Select Agent Program will provide a written decision granting or denying the request. Exclusions are effective upon notification to the applicant.

Excluded agents and toxins that have been granted an exclusion from the regulations can be found at http://www.selectagents.gov/SelectAgentsandToxinsExclusions.html. Until such time that exclusions are considered and granted, the attenuated strain of the select agent or select toxin that has been modified to be less potent or toxic is subject to the requirements of select agent regulations.
What information should an entity submit to the Federal Select Agent Program when requesting the exclusion of an "attenuated strain of a select agent or an inactive form of a toxin" from the requirements of the select agent regulations?
A cover letter signed by the entity's Responsible Official, on entity letterhead, requesting the exclusion of an attenuated strain of a select agent or inactive form of a select toxin. The request must contain the rational for exclusion of the strain or toxin and scientific references or supporting documentation that demonstrates the attenuated strain or toxin does not have the potential to pose a severe threat to public health and safety, animal or plant health, and/or animal or plant products. Some of the criteria used to determine if a strain or toxin should be excluded are given below. The entities request should contain information that is responsive to these criteria:
- Documented history of not causing disease in humans, or relevant animal or plant models.
- Defined genetic mutations or alterations known to attenuate virulence in humans or relevant animal or plant models.
- Data showing the mutations have a low frequency of reversion to wild-type virulence.
- Level of difficulty in engineering the attenuated strain to restore wild-type virulence. For each pathogen, the sample size and type of animal or plant model used to test virulence is important.
- Quantitative measures could be required, such as demonstration of a least 4-6 logs' reduction in virulence in an appropriate animal or plant model.
- Information regarding tests that may be conducted to differentiate animals or plants exposed to the attenuated strain from those infected with the wild-type organism.
- Related published scientific papers which support the methods and data provided for the exclusion.
- The data requirements necessary for consideration of an exclusion request for highly pathogenic Avian influenza (HPAI) virus are listed here.
I would like to submit a request to lower the biosafety containment for cDNA constructs derived from the Ebola virus and Marburg virus. How should I submit this request?

Currently, the cDNA of select agent viruses are not regulated under the select agent regulations. Therefore, the Federal Select Agent Program does not play a role in determining the biosafety containment of these genetic materials. However, the NIH Guidelines remain applicable to this and other research at institutions receiving NIH funding and performing work involving recombinant DNA (of certain human etiologic agents which are classified into groups according to their relative pathogenicity for healthy adult humans), including review and approval by an Institutional Biosafety Committee. Please refer to the following NIH Office of Biotechnology Activities guidance document for essential elements that should be included in the application to lower containment: "Points to Consider When Applying to the NIH Office of Biotechnology Activities (NIH OBA) for a Reduction in Biosafety Level (BL) Containment for Experiments Involving the Cloning of Full-length cDNA Constructs Derived from Risk Group (RG) 4 Viruses of the Order Mononegavirales".

For agriculture agents, please submit all biosafety containment questions to the Agriculture Select Agent Services (APHIS/USDA).
How long does it take for the Federal Select Agent Program to make a determination about an exclusion request?

The Federal Select Agent Program thoroughly reviews all exclusion requests. Some requests may require additional information from the entity before the review can proceed; therefore, the length of time varies considerably based upon the type of request proposed. On average, it takes 5-8 weeks for the Program to review the request and notify the entity of the decision. Entities registered with the Federal Select Agent Program may contact your file manager to follow up on the status of your request. Non-registered entities may contact the Federal Select Agent Program (lrsat@cdc.gov or AgSAS@aphis.usda.gov) to follow up on the status of your request.
Does an entity need to request prior approval to introduce antibiotic resistance into an excluded strain of a select agent?

If an entity modifies an excluded select agent such that virulence is restored or enhanced, then that select agent would be subject to all of the provisions of the select agent regulations including the pre-approval requirements of section 13 (Restricted Experiments). Please note that an attenuated strain of select agent that is excluded from the requirements of the select agent regulations is not exempt from other applicable regulations or guidelines (e.g., NIH guidelines, USDA/APHIS regulatory permits, etc.).
Are all attenuated strains of select agents or inactive forms of select toxins excluded from the regulation?

No. Only those attenuated strains of select agents or toxins that have been modified to be less potent or toxic that have been determined by the Federal Select Agent Program not to pose a severe threat to public health and safety, to animal and plant health, or to animal and plant products are excluded from the regulations. However, if an excluded attenuated strain or toxin that has been modified to be less potent or toxic is subjected to any manipulation that restores or enhances its virulence or toxic activity, the resulting agent or toxin will be subject to the requirements of 42 CFR Part 73, 9 CFR Part 121, and 7 CFR Part 331. Attenuated strains and inactive toxins that have been granted an exclusion from the regulations can be found at http://www.selectagents.gov/SelectAgentsandToxinsExclusions.html.

Exclusions – Toxins

The total amount of Staphylococcal Enterotoxin A purified in one experiment from a wild type S. aureus strain is below the regulated amount (100 mg). Is the toxin excluded from the select agent regulations?

It depends on the circumstance. The exclusion limit of a toxin is not determined per experiment, but determined by the aggregate volume of toxin produced, stored, and/or manipulated at any given time per principal investigator (PI). Therefore, if the PI controls more than 100 mg of Staphylococcal enterotoxins at any given time, the toxin is subject to the requirements of the select agent regulations.

Restricted Experiments

How do I submit a request for approval to conduct a restricted experiment?

A request for approval to conduct a restricted experiment should be submitted by the entity's Responsible Official and sent to the entity's Federal Select Agent Program file manager. The request will be reviewed by the Federal Select Agent Program. We recommend that it not be submitted as part of an amendment as doing so may delay approval of other changes proposed in the amendment while the restricted experiment request is under review.
What should an entity submit to the Federal Select Agent Program when requesting approval to conduct a restricted experiment?
A cover letter signed by the entity's Responsible Official, on entity letterhead, requesting approval to conduct the restricted experiment, along with the following information (if applicable):
- Synopsis of the proposed restricted experiment and the intended objective(s).
- Description of the nucleic acid insert (complete sequence information is not required) and the predicted biological characteristics of the recombinant or synthetic product.
- Description of the cloning/expression vector.
- Identification and characteristics of the host organism used for molecular cloning.
- Description of the methods used for selection (e.g. recombinant or passive selection) to include all potential drug resistant traits (including intermediate variants).
- Address whether there are alternative drug resistance marker genes that could be used (i.e., to avoid the acquisition of drug resistance that could compromise the use of the drug to control disease agents in humans, veterinary medicine, or agriculture). Provide an explanation as to why these alternative marker genes are not being considered.
- Description of biosafety level including facility containment, equipment and special practices to be utilized for the proposed restricted experiment(s).
- Description of the mechanism and specificity of the drug resistance (antibiotic and antiviral resistance) being conferred to include any cross-resistance to other therapeutically useful antimicrobials.
- Estimated amount of toxin (recombinant or synthetic) to be produced.
- Synopsis of any planned animal or plant experiments or other relevant animal or plant work.
- Scientific references or supporting documentation, particularly with respect to the biosafety aspects of the proposed experimental product.
Does the introduction of antimicrobial resistance into a select agent, to be used as a marker for cloning, meet the definition of a restricted experiment?

It depends on the circumstance. Although not all antibiotic resistance markers confer resistance to drugs used to control or prevent diseases associated with select agents and toxins, we highly recommend that all experiments involving the introduction of antimicrobial resistance into a select agent be reviewed by the Federal Select Agent Program to determine if the proposed experiment meets the definition of a restricted experiment. If the introduction of such a marker compromises the control of disease agents (e.g., in the event of an accidental exposure), it may meet the definition of a restricted experiment requiring prior approval. All restricted experiment requests are reviewed on a case-by-case basis.
Should intermediate selection markers not retained in the final construct be included in the request to perform a potential restricted experiment?

Yes. The request must describe all methods used for selection, including all intermediate vectors containing drug resistant markers or potential drug resistant traits. Refer to the Restricted Experiment Guidance Document for more information regarding submission of restricted experiment requests to the Federal Select Agent Program.
I would like to possess a product resulting from an approved restricted experiment. Am I required to submit a request to the Federal Select Agent Program in order to possess the product?

It depends on the circumstance. If the product of a restricted experiment is itself a select agent or toxin, the same approval to possess, use, and transfer the restricted experiment product is required under section 7 of the select agent regulations as for any other select agent or toxin. As to whether the product requires specific approval under section 13 of the regulations, it depends on when the product of the restricted experiment was created. If the product was created prior to December 4, 2012, the product of a restricted experiment does not require special approval for possession under section 13 of the select agent regulations. If the product was created on or after December 4, 2012, the possession is regulated under section 13 of the select agent regulations. For those regulated restricted experiment products, the entity must submit a written request to the Federal Select Agent Program asking permission to possess the product. If the possession of the product from a restricted experiment is approved, the entity must update the relevant information as an amendment to their select agent registration. In addition, the subsequent Principal Investigator (PI)'s work objective should be submitted for review, and the entity could expect that any approval would be subject to the same restrictions and biosafety requirements as were imposed by the initial "restricted experiment" approval. Please refer to the Restricted Experiment Guidance Document for more information regarding the regulation status of the possession of products from restricted experiments.
I would like to conduct a restricted experiment using passive selection (e.g., transfer of drug resistant traits through gradient or step-wise selective pressure) as a method to select for drug resistance traits in select agents. Am I required to submit a request to the Federal Select Agent Program to conduct this experiment?

Yes. The entity must submit a written request to the Federal Select Agent Program asking permission to conduct the experiment. If the request to perform the experiment is approved, the entity must update the relevant information as an amendment to their select agent registration. Only passive selection experiments conducted on or after December 4, 2012 are regulated under Section 13 of the select agent regulations. Please refer to the Restricted Experiment Guidance Document for more information regarding the regulation status to conduct restricted experiments using passive selection or utilizing recombinant DNA.
May an entity transfer the product of a restricted experiment to another PI?

Depending on when the product was generated, the possession of a product of a restricted experiment may require pre-approval from the Federal Select Agent Program before a transfer request is approved. Please refer to the Restricted Experiment Guidance Document for a list of documentation required for review.

In addition, the APHIS/CDC Form 2, Request to Transfer Select Agents and Toxins, must be completed and submitted to the Federal Select Agent Program prior to transferring any select agent(s) or any toxin(s) over the exclusion amount as indicated in section 3(d)(7). In Block 26, "characterization of agent" field, describe all drug resistance traits or recombinant DNA containing genes for the biosynthesis of select toxins, for each strain listed. Please refer to the instructions and guidance documents for Form 2 found at http://www.selectagents.gov/form2.html. If approved, note that an amendment to the recipient entity's registration for the receiving PI's work objective must be submitted to the Federal Select Agent Program.
I received approval from the Federal Select Agent Program to conduct a restricted experiment; however, I would like to modify the experiment. Should I submit a new request to the Federal Select Agent Program?

Yes. Restricted experiment decisions are determined by the specific work objectives and protocols submitted. If there are any changes in the work objectives or protocols, you should notify the Federal Select Agent Program and amend your select agent registration in accordance with section 7(h) of the select agent regulations.
How long does it take for the Federal Select Agent Program to make a final determination regarding a restricted experiment?

The Federal Select Agent Program thoroughly reviews all requests to conduct a restricted experiment. Some decisions can be determined quickly based on identical experiments that were previously reviewed by the program. Some requests may require additional information from the entity before the review can proceed. On average, it takes 5-8 weeks for the Federal Select Agent Program to review the request and notify the entity of the decision. Therefore, the length of time varies considerably based upon the type of request proposed. You may contact your file manager to follow up on the status of your request.
An entity submits a request to: 1) Perform an experiment that involves the deliberate formation of synthetic or recombinant DNA containing genes for the biosynthesis of a select toxin lethal for vertebrates at an LD[50] < 100 ng/kg body weight; and 2) Introduce synthetic or recombinant nucleic acids that encode for the functional form of a select toxin into an E. coli strain. The propagation of the strain will not produce toxin that exceeds the regulated amount. Do the experiments meet the definition of a restricted experiment?

Experiment 1:
Yes, the experiment involves the deliberate formation of synthetic or recombinant DNA containing genes for the biosynthesis of a select toxin if the select toxin is lethal for vertebrates at an LD[50]<100ng/kg body weight. A formal request to perform this work should be submitted to the Federal Select Agent Program.

Experiment 2:
No, this is not considered a restricted experiment, but the construct (product of the experiment) is subject to the select agent regulations. Although wild-type E.coli strains alone are not considered select agents and the expected yield of toxin is less than the regulated amount, since the recombinant E. coli strain contains the nucleic acids that encode for and express a functional form of the toxin, the recombinant strain is now considered a select agent and subject to select agent regulations. This work must be included in the registration application.
Section 13 of the select agent regulations states “…Experiments involving the deliberate formation of synthetic or recombinant DNA containing genes for the biosynthesis of select toxins lethal for vertebrates at an LD[50] < 100 ng/kg body weight…” Which of the select toxins meet this definition?

Botulinum neurotoxin is the only select toxin that meets this definition.