Appendix D: Vertebrate Animal Biosafety Level 2 (ABSL-2) Requirements
*IACUC, Laboratory Animal Resources (LAR) Policies and Procedures must be followed for ABSL-2 work
This section outlines the biosafety standards and operational requirements for research involving vertebrate animals that are experimentally infected with, or exposed to, Risk Group 2 (RG2) agents or materials of human origin at the Animal Biosafety Level 2 (ABSL-2) containment level. The intent is to ensure the protection of personnel, animals, and the environment in accordance with the BMBL (6th Edition), NIH Guidelines, and University of Montana biosafety and animal care policies.
ABSL-2 facilities are designed for work with agents associated with human disease that present moderate hazards to personnel and the environment. This level of containment provides safeguards against infection through ingestion, inoculation, and mucous membrane exposure. All ABSL-2 research must be reviewed and approved by both the Institutional Biosafety Committee (IBC) and the Institutional Animal Care and Use Committee (IACUC) prior to initiation.
Institutional Oversight and Access Control
The animal facility Manager, principal investigator (PI), and biosafety staff share responsibility for ensuring that appropriate biosafety, biosecurity, and emergency procedures are implemented and enforced. Access to ABSL-2 areas is restricted to trained and authorized personnel whose entry is required for research, animal care, or support activities.
Each protocol involving infectious or potentially infectious materials must be reviewed through the IBC and IACUC to ensure that specific biohazards related to the agent and animal species are addressed. Access to animal rooms is controlled, and signage indicating the Animal Biosafety Level, responsible personnel, emergency contact information, and required personal protective equipment (PPE) must be clearly posted when infectious agents are in use.
Training and Medical Surveillance
All personnel working in ABSL-2 facilities must receive comprehensive training on biosafety principles, potential hazards, exposure prevention, and emergency response. Training is required prior to beginning work and refreshed at least annually, or whenever new hazards or procedures are introduced.
Personnel must also be enrolled in the University’s Occupational Health and Safety Program. This includes medical evaluations, allergy screening, and immunizations or prophylaxis when appropriate. Individuals with health conditions that may increase susceptibility to infection are advised to consult Occupational Health for individualized assessment and counseling.
Operational and Work Practices
All activities involving infectious materials must be conducted with care to prevent exposure and the creation of aerosols or splashes. Procedures likely to generate aerosols should be performed inside a certified Biological Safety Cabinet (BSC) or other approved containment device. Eating, drinking, applying cosmetics, and handling contact lenses are prohibited in animal areas. Sharps use is minimized whenever possible. Needles and syringes are used only when necessary, and safer sharps devices must be implemented when feasible. Needles must never be recapped or removed by hand; instead, they are placed directly into puncture-resistant sharps containers located near the point of use. Personnel must wash their hands after handling animals, infectious materials, or cages, and before leaving the containment area. Long hair should be restrained, and gloves must be worn at all times when handling animals or contaminated equipment.
Decontamination and Waste Management
Work surfaces must be decontaminated after each procedure and following any spill using an effective disinfectant appropriate for the agent in use. All potentially infectious materials, carcasses, bedding, and disposable equipment must be decontaminated—typically by autoclaving—before disposal or removal from the facility. Equipment and caging must be decontaminated prior to washing, repair, or maintenance. In the event of gross contamination or facility renovation, complete room decontamination must be performed. Spills or exposure incidents must be reported immediately to the PI and Biosafety Officer, and response procedures must follow institutional emergency protocols.
Safety Equipment and Personal Protective Equipment (PPE)
All manipulations of infectious materials are performed within a certified Biological Safety Cabinet or equivalent containment device. Infected animals should be housed in appropriate primary containment systems, such as solid-bottom cages with microisolator lids or ventilated cage racks with HEPA filtration.
Personnel must wear appropriate protective clothing, including lab coats, gowns, gloves, and eye protection. Additional PPE—such as sleeve covers, respirators, or face shields—may be required based on the risk assessment. All PPE is to be removed prior to exiting the containment area and must not be worn outside the facility. Reusable clothing is laundered by the institution; it must never be taken home.
Facility Design and Engineering Controls
ABSL-2 animal facilities must be physically separated from general traffic areas and equipped with self-closing, lockable doors. The entryway should include a handwashing sink and eyewash station located near the exit. If required by risk assessment, showers should be available within or adjacent to the facility.
Walls, floors, and ceilings must be sealed, water-resistant, and easily cleanable. Floors should slope toward drains with appropriate traps or disinfectant seals. Windows, if present, must be sealed and resistant to breakage.
Ventilation systems must maintain directional airflow into the ABSL-2 area, with exhaust air discharged to the outside and not recirculated. HEPA filtration is required for exhaust air from ventilated cage racks and BSCs. Cage washers must achieve a final rinse temperature of at least 180°F, and an autoclave or validated alternative must be available for the decontamination of waste and carcasses.
Incident Response and Reporting
All exposures, bites, scratches, spills, or potential contamination incidents must be reported immediately to supervisory and biosafety personnel. Written incident reports must be submitted within 24 hours. Appropriate medical evaluation and follow-up are coordinated through the University’s Occupational Health Program.