Select Agent and Toxin Regulations: Beyond the Eighth Edition of the Guide for the Care and Use of Laboratory Animals

Abstract

In the interval between the publication of the seventh and eighth editions of the Guide for the Care and Use of Laboratory Animals (Guide), much has changed with regard to the regulation and funding of highly pathogenic biologic agents and toxins (Select Agents). Funding of research involving highly pathogenic agents has increased dramatically during this time, thus increasing the demand for facilities capable of supporting this work. The eighth edition of the Guide briefly mentions Select Agents and provides a limited set of references. Here we provide some background information regarding the relevant laws and regulations, as well as an overview of the programmatic requirements pertaining to the use of Select Agents, with a focus on use in animals.

The eighth edition of the Guide for the Care and Use of Laboratory Animals (the Guide) states that “use of highly pathogenic ‘select agents and toxins’ in research requires that institutions develop a program and procedures for procuring, maintaining, and disposing of these agents.”¹³ In addition to this statement in the second chapter, the hazardous agent containment section in Chapter 5 mentions the need to implement enhanced security and follow applicable regulations when using certain biologic agents and toxins. The legislation establishing increased oversight and restrictions for certain biologic agents was enacted the same year as the publication of the seventh edition of the Guide.¹²An exponential increase in research funding for bioterrorism agents has occurred since the publication of the seventh edition of the Guide, leading to a need for a greater discussion of the requirements beyond what is mentioned in the eighth edition.^7,9,15 The fifth edition of the Biosafety in Microbiologic and Biomedical Laboratories manual also provides information, greatly expanded from the fourth edition, on the regulation, use, and handling of Select Agents and Toxins.²⁸

Definitions

A recent presidential Executive Order defined Biologic Select Agents and Toxins as “biological agents and toxins with the potential to pose a severe threat to public health and safety, animal and plant health, or animal and plant products and whose possession, use, and transfer are regulated by the Department of Health and Human Services (DHHS) and the Department of Agriculture under the [Select Agent Regulations].”²⁷Select Agents or Toxins include the 39 organisms or toxins that the DHHS currently has designated as posing substantial harm to human health²⁰ (Figure 1). A separate list is maintained by the United States Department of Agriculture (USDA) for the 31 Select Agents and Toxins that are considered to pose a severe threat to the health of animals or plants⁴ (Figure 1). The 10 zoonotic agents or toxins that the USDA and DHHS deem as a threat to both animal health and public health are identified as Overlap Agents and can be regulated by either agency (Figure 1). These lists are reviewed biennially by a convened committee and have been amended over the years; the lists are undergoing review currently.^5,21,22

Figure 1.

Currently regulated Select Agents and Toxins, with historical amendments. Adapted from reference 16. a, Formerly Cercopithecine herpesvirus 1;¹² b, changed from Overlap Agent to being regulated by CDC.²

Regulatory Framework

The 1996 Antiterrorism and Effective Death Penalty Act required that the Secretary of Health and Human Services regulate any “biological agent that has the potential to pose a severe threat to public health and safety.”⁶ The subsequent regulations, issued through DHHS, focused on the transfer and transport of designated agents between laboratories and disposal of agents, as well as registration of laboratories containing certain biologic agents.¹ The laboratories were directed to follow prior guidance, such as the Biosafety in Microbiologic and Biomedical Laboratories manual,²⁸ for handling the agents and toxins. Of the 18 agents designated for greatest oversight, 2 (Korean Hemorrhagic Fever virus and Histoplasma capsulatum) have been removed from Select Agent status in subsequent regulations.

The passage of the USA PATRIOT (Uniting and Strengthening America by Providing Appropriate Tools Required to Intercept and Obstruct Terrorism) Act in 2001 and the Public Health Security and Bioterrorism Preparedness and Response Act in 2002 established the regulations discussed in the eighth edition of the Guide.^25,27 The USA PATRIOT Act expanded the 1996 statute to add penalties for the possession of biologic agents, toxins, or delivery systems if not being used for a legal peaceful purpose. In addition, this law defined a “restricted person” who is prohibited from transporting or possessing a Select Agent.²⁷

In addition to developing and implementing various preparedness strategies and protecting food, drug, and water supplies, the 2002 Public Health Security and Bioterrorism Preparedness and Response Act directed the Secretary of Health and Human Services and the Secretary of Agriculture to establish and maintain a list of regulated agents and toxins and develop regulations. These regulations are intended to ensure that procedures are in place to safeguard the agents and toxins during their transfer and use, including identifying personnel who have access to the agents, preventing restricted persons from gaining access, and ensuring that personnel handling the agents and toxins are appropriately trained and skilled to conduct the work. The law also discussed coordination of the Overlap Agents that can be regulated by either agency to minimize any regulatory conflict and decrease administrative burden. The law delineated the notification and procedures to be followed in the event of a release of a biologic agent or toxin.

In 2005, DHHS and the USDA concurrently published regulations implementing the 2002 Act to protect public health and safety.^2,18 The DHHS Final Rule designated the Centers for Disease Control and Prevention (CDC) for implementation of the provisions and identified 21 regulated agents and toxins (Figure 1). The USDA regulations are implemented through the Animal and Plant Health Inspection Service (APHIS) and cover 31 plant or animal pathogens (Figure 1). In addition, the 2005 regulations designated 20 agents or toxins as Overlap Agents that could be regulated by either entity. The regulations delineate exemptions, institutional and regulatory oversight, civil and criminal penalties, and the requirements for agent inventory, security, biosafety, emergency response, training, inspections, transfers, and transportation. The regulations focus on performance standards that are based on the risks associated with a defined agent with a specified use in a particular location. In 2008, both the USDA and DHHS updated their Final Rules after a biennial review of the agents and toxins listed. Accordingly 10 agents or toxins were shifted from the list of Overlap Agents, capable of regulation by either USDA or CDC, to regulation solely by the CDC (Figure 1).

Implementing the Regulations for Animal Studies

The first consideration in determining whether a planned study is subject to regulation is identifying whether a regulated agent or toxin will be used (Figure 2). Animals inoculated with Select Agents, including botulinum-neurotoxin–producing strains of Clostridium, are regulated as Select Agents. Although animals inoculated with Select Toxins are not regulated or required to be housed in a registered space, the Select Toxin must be handled according to regulations until the toxin is in the animal. Therefore, any areas in which animals are exposed to Select Toxins or where the toxins are stored prior to the exposures must be registered. Agents or toxins in their naturally occurring environment, nonviable select agents, and nonfunctional toxins are excluded from the regulations. An example of a Select Agent that is exempt from regulation because it is in its natural environment is B virus (Macacine herpesvirus 1)¹⁴ in a macaque monkey that had not been infected intentionally. In addition, a list of excluded strains of Select Agents is published on the National Select Agent Registry.¹⁶ These strains are exempt from regulations if used for certain activities, including basic or applied research. Also included among the regulatory exemptions are clinical or diagnostic laboratories performing certain well-defined activities listed in the regulations. However, the identification of a Select Agent or Toxin in a diagnostic sample must be reported appropriately within a designated timeframe, depending on the agent identified. The National Select Agent Registry has set forward examples detailing when some of these reporting requirements apply. For example, if a Select Agent is diagnosed in an animal presented for necropsy or care, and personnel potentially were exposed during the process, then the facility must notify the CDC and APHIS about the release of a Select Agent. The examples clarifying this rule involve the necropsy of a deer which later is identified as infected with Eastern equine encephalitis virus and the care of a cat, from which Yersinia pestis was isolated subsequently, by a veterinary clinic. Both the necropsy facility and veterinary clinic that isolated these agents from the infected animals are required to report the exposures as the release of a Select Agent. However, a bite or scratch inflicted on animal care personnel from an Old World primate that subsequently is confirmed to be positive for Macacine herpesvirus 1 (B virus) is not reportable.

Figure 2.

Factors to consider when determining whether a proposed animal study meets the definition of a study using a Select Agent or Toxin.

The use of a regulated Select Agent in an animal requires a Certificate of Registration. This certification must be granted by the appropriate agency (CDC or APHIS) before the agent is obtained, used, or transferred. Designation of the Responsible Official and a description of the activities and agents or toxins to be used are included in this registration. The Responsible Official must be familiar with the regulations, obtain a Security Risk Assessment, and be empowered to act on behalf of the entity to ensure compliance with the regulations. The CDC also recommends that the Responsible Official be free of potential conflicts of interest, such as directly working with regulated agents or toxins.¹¹ Furthermore, the Responsible Official is the primary point of contact for the entity with the CDC and APHIS. The issuance of a Certificate of Registration may be contingent on an inspection or review of security, biosafety, incident response, or other associated plans. The registration, once issued, is valid for a maximum of 3 y. The National Select Agent Registry website contains the registration forms, training materials, and checklists that are helpful for becoming registered. The registration process may take several months.¹⁷

The Security Risk Assessment is conducted through the Federal Bureau of Investigation to ensure that restricted persons do not gain access to the agents and toxins. The Security Risk Assessment is specific to the person, is nontransferrable between entities, must be renewed every 3 y, and must be completed and approved before a person can have access to a regulated agent or toxin. Persons who are required to obtain a Security Risk Assessment include the Responsible Official, scientists handling the agent, and animal care personnel handling animals infected with Select Agents. To receive a Security Risk Assessment, the person must have a legal presence in the United States (citizenship, ‘green card’) and submit the appropriate federal form and 2 sets of fingerprints to the Federal Bureau of Investigation. This information then is used to search a wide variety of criminal, immigration, and national security databases.⁸ The Federal Bureau of Investigation report assists the agency responsible for regulating the agents being used in determining whether the person should be permitted access to the designated select agent at the prescribed location for the defined use. Currently the APHIS Administrator is allowed to permit a restricted person to have limited access to Select Agents that are regulated solely by the USDA.²⁴ Persons who are designated as restricted persons are informed of the information that conferred such status and may appeal this designation by filing a request for reassessment. The Federal Bureau of Investigation will reinvestigate and, depending on their findings and any documentation submitted at the time of the appeal, will determine whether there is sufficient evidence to declare a reversal of the restricted person designation. The regulated entity may impose additional requirements beyond the Security Risk Assessment for unescorted access.

Implementing Biosecurity

A multiagency working group report recently defined biosecurity as “the protection, control of, and accountability for high-consequence biological agents and toxins, and critical relevant biological materials and information within laboratories to prevent unauthorized possession, loss, theft, misuse, diversion, or intentional release”²⁴. Similarly, the Biosafety in Microbiologic and Biomedical Laboratories manual refers to biosecurity as “the protection of microbial agents from loss, theft, diversion or intentional misuse.”²⁸ The National Select Agent Registry posts on their website various checklists and templates for security and incident response plans that are useful in developing a biosecurity program, which should include access control, physical security, and transportation. Any security, biocontainment, and incident response plans must be tested annually and updated as needed, especially when areas are identified for improvement through these drills. In addition, the plans must include provisions for escort of visitors by persons with approved Security Risk Assessments, as well as access control if keys or passwords are lost or compromised or during times of staff turnover. A visitor is considered to be any person that has not been granted access to a Select Agent or Toxin by the USDA or CDC for a specified location and agent. Visitors must be escorted by an approved person at all times, and a log must be generated with the names of the visitor and escort and the date and time of entry into the secured area. In addition, visitors should receive appropriate area- and agent-specific training before entering the secured area. Persons who have been granted unescorted access at one entity and are visiting a different entity require an amendment to the new entity's registration for unescorted access at the different entities.

Long-Term Storage of Specimens

Samples from animals need not be tracked as long-term storage when there is no intent to store the samples for a prolonged period or when a date is specified by which time the sample will be used. In addition, a process must be in place for the identification of samples that were obtained from animals inoculated with Select Agents or Toxins but that are shown to be free of the agent or contain only inactivated or attenuated agent and thus are no longer subject to regulation. Samples that do not meet these criteria must be handled as for long-term storage and maintained in a secure location, along with a detailed, accurate inventory for each sample that is placed into long-term storage. This inventory must identify the agent and include the source, quantity, storage site, purpose of use, and additional information, such as tracking the movement of aliquots removed and returned to the storage site.

Financial Impact

The CDC economic analysis estimated that annualized costs for implementing the regulations ranged from approximately $15,000 to more than $150,000 for each location.¹⁹ The USDA analysis is in agreement with these costs and provides an estimate of $50,000 for ongoing expenditures.³ The largest annual costs are predicted to be associated with maintaining physical security of the facilities, performing agent and toxin inventories, and administrative requirements. These costs typically are in addition to any construction costs of building to the appropriate biocontainment level.^10,15

Future Directions

President Obama issued an Executive Order in 2010 that led to the creation of the Federal Experts Security Advisory Panel (Figure 3) to advise DHHS and USDA in designating a subset of regulated agents, Tier 1, that pose the highest risk for misuse with a resultant significant impact.²⁶ The Executive Order also stated that a reduction in the total number of agents and toxins on the list should be considered, as part of a risk-based tiering. The 13 June 2011 report from the Federal Experts Security Advisory Panel that was released in response to this Executive Order proposed 11 biologic agents and toxins listed in Tier 1 and 25 agents, toxins, or select strains to be deleted from the list.²³ These recommendations must still be formalized into regulations through a public comment period that closed in January 2012.^5,21 The proposed regulations differ slightly from the Executive Order with the addition of 2 arenaviruses for regulation (Lujo and Chapare viruses) and the removal of 17 agents (Figure 4). The advisory panel also recommended increasing the available guidance for addressing common issues and preventing inadvertent noncompliance.

Figure 3.

The departments, agencies, and offices represented on the Federal Experts Security Advisory Panel, cochaired by members from the Department of Health and Human Services and the US Department of Agriculture.

Figure 4.

Recommendations from the Federal Experts Security Advisory Panel for Tier 1 agents and those agents that should be considered for removal from the regulated Select Agents and Toxins list. a, Certain subtypes and genotypes only.

Other future considerations for the laboratory animal community are the identification of personnel who not only have experience with the care of the animals but who also are able to obtain clearance for unescorted access to the animals inoculated with Select Agents.¹⁴ The American Association for Laboratory Animal Science and the American Biologic Safety Association have begun to offer training resources to the biomedical community.

Summary

The development of a biosecurity program to house research animals inoculated with Select Agents or Toxins may appear daunting. Understanding the regulations and being aware of available resources can facilitate obtaining a Certificate of Registration. Ongoing discussions among members of the laboratory animal community are needed to share best practices, especially in training and retaining staff qualified to work with Select Agents or Toxins in animals.