Last update 01/05/2008

The following three combinations of practices, safety equipment, and facilities are used for experiments on animals infected with agents which produce, or may produce, human infection. They provide increasing levels of protection to personnel and to the environment, and are recommended as minimal standards for activities involving infected laboratory animals. These practices and procedures apply to animal projects not involving recombinant DNA molecules. The biosafety level requirements for rDNA animal research are listed in Appendix C.

These three combinations, designated Animal Biosafety Levels (ABL) 1 to 3, describe animal facilities and practices applicable to work on animals infected with agents assigned to the corresponding Risk Groups 1-3.

Work with Risk Group 4 agents or projects requiring ABL-4 are not permitted at OSU.

Animal Biosafety Level 1

Standard Practices (ABL-1)

• Access to the animal facility is limited or restricted at the discretion of the laboratory or animal facility director/PI.
• Personnel wash their hands after handling cultures and animals, after removing gloves, and before leaving the animal facility.
• Eating, drinking, smoking, handling contact lenses, applying cosmetics, and storing food for human use are not permitted in animal rooms. Persons who wear contact lenses in animal rooms should also wear goggles or a face shield.
• All procedures are carefully performed to minimize the creation of aerosols. Work surfaces are decontaminated after use or after any spill of viable materials.
• Doors to animal rooms open inward, are self-closing and are kept closed when experimental animals are present.
• All wastes from the animal room are appropriately decontaminated, preferably by autoclaving, before disposal. Infected animal carcasses are incinerated after being transported from the animal room in leakproof, covered containers.
• An insect and rodent control program is in effect. Contact Pest Management 7-4038 for more information.

Special Practices (ABL-1)

• The laboratory or animal facility director in coordination with the PI limits access to the animal room to personnel who have been advised of the potential hazard and who need to enter the room for program or service purposes when work is in progress. In general, persons who may be at increased risk of acquiring infection, or for whom infection might be unusually hazardous, are not allowed in the animal room.
• The laboratory or animal facility director in coordination with the PI establishes policies and procedures whereby only persons who have been advised of the potential hazard and meet any specific requirements (e.g., immunization) may enter the animal room.
• Bedding materials form animal cages are removed in such a manner as to minimize the creation of aerosols, and are disposed of in compliance with applicable institutional or local requirements.
• Cages are washed manually or in the cage washer. Temperature of final rinse water in a mechanical washer should be 180° F.
• The wearing of laboratory coats, gowns, or uniforms in the animal facility is recommended. It is further recommended that laboratory coats worn in the animal facility not be worn in other areas.
• OSU's biosafety manual is prepared or adopted. Personnel are advised of special hazards, and are required to read and to follow instructions on practices and procedures outlined in the biosafety manual

Safety Equipment (Primary Barriers, ABL-1)

• Special containment equipment is not required for animals infected with agents assigned to Biosafety level 1.

Animal Facilities (Secondary Barriers, ABL-1)

• The animal facility is designed and constructed to facilitate cleaning and housekeeping.
• A hand washing sink is available in the room where infected animals are housed.
• If the animal facility has windows that open, they are fitted with fly screens.
• Exhaust air is discharged to the outside without being recirculated to other rooms, and it is recommended, but not required, that the direction of airflow in the animal facility is inward.
• An autoclave which can be used for decontaminating infectious laboratory waste is available in the building with the animal facility.

Animal Biosafety Level 2

Standard Practices (ABL-2)

• Access to the animal facility is limited or restricted at the discretion of the laboratory or animal facility director/PI.
• Personnel wash their hands after handling cultures and animals, after removing gloves, and before leaving the animal facility.
• Eating, drinking, smoking, handling contact lenses, applying cosmetics, and storing food for human use are not permitted in animal rooms. Persons who wear contact lenses in animal rooms should also wear goggles or a face shield.
• All procedures are carefully performed to minimize the creation of aerosols. Work surfaces are decontaminated after use or after any spill of viable materials.
• Doors to animal rooms open inward, are self-closing and are kept closed when experimental animals are present.
• All wastes from the animal room are appropriately decontaminated, preferably by autoclaving, before disposal. Infected animal carcasses are incinerated after being transported from the animal room in leakproof, covered containers.
• An insect and rodent control program is in effect. Contact Pest Management at 7-4038 for more information.

Special Practices (ABL-2)

• The animal facility manager in coordination with the PI limits access to the animal room to personnel who have been advised of the potential hazard and who need to enter the room for program or service purposes when work is in progress. In general, persons who may be at increased risk of acquiring infection, or for whom infection might be unusually hazardous, are not allowed in the animal room.
• The animal facility manager in coordination with the PI establishes policies and procedures whereby only persons who have been advised of the potential hazard and meet any specific requirements (e.g., immunization) may enter the animal room.
• All areas operating at ABL-2 need to have the universal biohazard symbol attached to the main entrance door(s). When the infectious agent(s) in use in the animal room requires special entry provisions (e.g., the need for immunizations and respirators) an AUCAUC approved Animal Use Form is posted on the access door to the animal room. This form identifies the infectious agent(s) in use, lists the name and telephone number of the project supervisor or other responsible person(s), and indicates the special requirement(s) for entering the animal room as well as any other necessary procedures and practices.
• Laboratory personnel receive appropriate immunizations or tests for the agents handled or potentially present in the laboratory (e.g., hepatitis B vaccine or TB skin testing).
• When appropriate, considering the agents handled, baseline serum samples from animal care and other at-risk personnel may be collected and stored. Additional serum samples may be collected periodically depending on the agents handled or the function of the facility. The decision to establish a serologic surveillance program must take into account the availability of methods for the assessment of antibody to the agent(s) of concern. The program should provide for the testing of serum samples at each collection interval and the communication of results to the participants.
• OSU’s biosafety manual is adopted. Personnel are advised of special hazards, and are required to read and to follow instructions on practices and procedures outlined in the biosafety manual.
• Laboratory personnel receive appropriate training through the PI on the potential hazards associated with the work involved, the necessary precautions to prevent exposures, and the exposure evaluation procedures. Personnel receive annual updates, or additional training as necessary for procedural or policy changes. In addition, attendance at the ORCBS biosafety training is recommended.
• A high degree of precaution must always be taken with any contaminated sharp items, including needles and syringes, slides, pipettes, capillary tubes, and scalpels. Needles and syringes or other sharp instruments are restricted in the animal facility for use only when there is no alternative, such as for parenteral injection, blood collection, or aspiration of fluids from laboratory animals and diaphragm bottles. Plasticware should be substituted for glassware whenever possible.
• Only needle-locking syringes or disposable syringe-needle units (i.e., needle is integral to the syringe) are used for injection or aspiration of infectious materials. Used disposable needles must not be bent, sheared, broken, recapped, removed from disposable syringes, or otherwise manipulated by hand before disposal; rather, they must be carefully placed in conveniently located OSU approved, "Sharps" containers. Non-disposable sharps must be placed in a hard-walled container for transport to a processing area for decontamination, preferably by autoclaving.
• Syringes which re-sheathe the needle, needle-less systems, and other safe devices should be used when appropriate.
• Broken glassware must not be handled directly by hand, but must be removed by mechanical means such as a brush and dustpan, tongs, or forceps and disposed of in Sharps containers. Sharps containers filled 2/3 need to be disposed of and will be picked-up by EH&S for further processing. For more information refer to the Biohazardous Waste section.
• Cultures, tissues, or specimens of body fluids are placed in a container that prevents leakage during collection, handling, processing, storage, transport, or shipping. For more information refer to the Transportation of Biological Materials section.
• Cages are appropriately decontaminated, preferably by autoclaving, before they are cleaned and washed. Equipment and work surfaces should be decontaminated with an appropriate disinfectant on a routine basis, after work with infectious materials is finished, and especially after overt spills, splashes, or other contamination by infectious materials. Contaminated equipment must be decontaminated according to any local, state, or federal regulations before it is sent for repair or maintenance or packaged for transport in accordance with applicable local, state, or federal regulations, before removal from the facility.
• Spills and accidents which result in overt exposures to infectious materials are immediately reported to the laboratory director or PI. Medical evaluation, surveillance, and treatment are provided as appropriate and written records are maintained.
• Animals not involved in the work being performed are not permitted in the lab.

Safety Equipment (Primary Barriers, ABL-2)

• Biological safety cabinets, other physical containment devices, and/or personal protective equipment (e.g., respirators, face shields) are used whenever procedures with a high potential for creating aerosols are conducted. These include necropsy of infected animals, harvesting of tissues or fluids from infected animals or eggs, intranasal inoculation of animals, and manipulations of high concentrations or large volumes of infectious materials.
• Appropriate face/eye and respiratory protection is worn by all personnel entering animal rooms housing non-human primates.
• Laboratory coats, gowns, or uniforms are worn while in the animal room. This protective clothing is removed before leaving the animal facility.
• Special care is taken to avoid skin contamination with infectious materials. Gloves are worn when handling infected animals and when skin contact with infectious materials is unavoidable.

Animal Facilities (Secondary Barriers, ABL-2)

• The animal facility is designed and constructed to facilitate cleaning and housekeeping.
• A hand washing sink is available in the room where infected animals are housed.
• If the animal facility has windows that open, they are fitted with fly screens.
• If floor drains are provided, the drain traps are always filled with water or a suitable disinfectant.
• Exhaust air is discharged to the outside without being recirculated to other rooms, and it is recommended, but not required, that the direction of airflow in the animal facility is inward.
• An autoclave which can be used for decontaminating infectious laboratory waste is available in the building with the animal facility.

Animal Biosafety Level 3

Standard Practices (ABL-3)

• Access to the animal facility is limited or restricted at the discretion of the laboratory or animal facility manager or director/PI.
• Personnel wash their hands after handling cultures and animals, after removing gloves, and before leaving the animal facility.
• Eating, drinking, smoking, handling contact lenses, applying cosmetics, and storing food for human use are not permitted in animal rooms. Persons who wear contact lenses in animal rooms should also wear goggles or a face shield.
• All procedures are carefully performed to minimize the creation of aerosols.
• Work surfaces are decontaminated after use or after any spill of viable materials.
• Doors to animal rooms open inward, are self-closing and are kept closed when experimental animals are present.
• All wastes from the animal room are appropriately decontaminated, preferably by autoclaving, before disposal. Infected animal carcasses are incinerated after being transported from the animal room in leakproof, covered containers.
• An insect and rodent control program is in effect. Contact Pest Management at 7-4038 for more information.

Special Practices (ABL-3)

• The facility manger in coordination with the PI or other responsible person restricts access to the animal room to personnel who have been advised of the potential hazard and who need to enter the room for program or service purposes when infected animals are present. Persons who are at increased risk of acquiring infection, or for whom infection might be unusually hazardous, are not allowed in the animal room. Persons at increased risk may include children, pregnant women, and persons who are immunodeficient or immunosuppressed. The facility manager has the final responsibility for assessing each circumstance and determining who may enter or work in the facility.
• The facility manger in coordination with the PI establishes policies and procedures whereby only persons who have been advised of the potential hazard and meet any specific requirements (e.g., immunization) may enter the animal room.
• All areas operating at ABL-3 need to have the universal biohazard symbol (Figure 1) attached to the main entrance door(s) When the infectious agent(s) in use in the animal room requires special entry provisions (e.g., the need for immunizations and respirators) an AUCAUC approved Animal Use Form (Appendix G) is posted on the access door to the animal room. This form identifies the infectious agent(s) in use, lists the name and telephone number of the project supervisor or other responsible person(s), and indicates the special requirement(s) for entering the animal room as well as any other necessary procedures and practices.
• Laboratory personnel receive appropriate immunizations or tests for the agents handled or potentially present in the laboratory (e.g., hepatitis B vaccine or TB skin testing).
• Baseline serum samples from all personnel working in the facility and other at-risk personnel maybe collected and stored. Additional serum samples may be collected periodically and stored. If initiated, the serum surveillance program must take into account the availability of methods for the assessment of antibody to the agent(s) of concern. The program should provide for the testing of serum samples at each collection interval and the communication of results to the participants.
• OSU’s biosafety manual is adopted. Personnel are advised of special hazards, and are required to read and to follow instructions on practices and procedures outlined in the biosafety manual.
• Laboratory personnel receive appropriate training through the PI and the facility manager on the potential hazards associated with the work involved, the necessary precautions to prevent exposures, exposure evaluation procedures and emergency procedures. Personnel receive annual updates, or additional training as necessary for procedural or policy changes. In addition, attendance at the EH&S biosafety training is mandatory for ABL-3 work.
• A high degree of precaution must always be taken with any contaminated sharp items, including needles and syringes, slides, pipettes, capillary tubas, and scalpels. Needles and syringes or other sharp instruments are restricted in the animal facility for use only when there is no alternative, such as for parenteral injection, blood collection, or aspiration of fluids from laboratory animals and diaphragm bottles. Plasticware should be substituted for glassware whenever possible.
• Only needle-locking syringes or disposable syringe-needle units (i.e., needle is integral to the syringe) are used for injection or aspiration of infectious materials. Used disposable needles must not be bent, sheared, broken, recapped, removed from disposable syringes, or otherwise manipulated by hand before disposal; rather, they must be carefully placed in conveniently located OSU approved, "Sharps" containers. Non-disposable sharps must be placed in a hard-walled container for transport to a processing area for decontamination, preferably by autoclaving.
• Syringes which re-sheathe the needle, needle-less systems, and other safe devices should be used when appropriate.
• Broken glassware must not be handled directly by hand, but must be removed by mechanical means such as a brush and dustpan, tongs, or forceps and disposed of in Sharps containers. Sharps containers filled 2/3 need to be disposed of and will be picked-up by EH&S for further processing. For more information refer to the Biohazardous Waste section.
• Cultures, tissues, or specimens of body fluids are placed in a container that prevents leakage during collection, handling, processing, storage, transport, or shipping. For more information refer to the Transportation of Biological Materials section.
• Cages are appropriately decontaminated, preferably by autoclaving, before they are cleaned and washed. Equipment and work surfaces should be decontaminated with an appropriate disinfectant on a routine basis, after work with infectious materials is finished, and especially after overt spills, splashes, or other contamination by infectious materials. Contaminated equipment must be decontaminated and accompanied by OSU’s Equipment Release Form (Appendix H) before it is sent for repair or maintenance or packaged for transport in accordance with applicable local, state, or federal regulations, before removal from the facility.
• Spills and accidents which result in overt exposures to infectious materials are immediately reported to the laboratory director, facility manger (if applicable) and EH&S. Medical evaluation, surveillance, and treatment are provided as appropriate and written records are maintained.
• All wastes from the animal room are autoclaved before disposal. All animal carcasses are incinerated. Dead animals are transported from the animal room to the incinerator in leakproof covered containers.
• Animals not involved in the work being performed are not permitted in the lab.

Safety Equipment (Primary Barriers, ABL-3)

• Personal protective equipment is used for all activities involving manipulations of infectious materials or infected animals.
• Wrap-around or solid-front gowns or uniforms are worn by personnel entering the animal room. Front-button laboratory coats are unsuitable. Protective gowns should be appropriately contained until decontamination or disposal.
• Personnel wear gloves when handling infected animals. Gloves are removed aseptically and autoclaved with other animal room wastes before disposal.
• Appropriate face/eye and respiratory protection is worn by all personnel entering animal rooms housing non-human primates.
• Boots, shoe covers, or other protective footwear, and disinfectant footbaths are available and used when indicated.
• Physical containment devices and equipment appropriate for the animal species are used for all procedures and manipulations of infectious materials or infected animals.
• The risk of infectious aerosols from infected animals or their bedding also can be reduced if animals are housed in partial containment caging systems, such as open cages placed in ventilated enclosures (e.g., laminar flow cabinets), solid wall and bottom cages covered with filter bonnets, or other equivalent primary containment systems.

Animal Facilities (Secondary Barriers, ABL-3)

• The animal facility is designed and constructed to facilitate cleaning and housekeeping, and is separated from areas which are open to unrestricted personnel traffic within the building. Passage through two sets of doors is the basic requirement for entry into the animal room from access corridors or other contiguous areas. Physical separation of the animal room from access corridors or other activities may also be provided by a double-doored clothes change room (showers may be included), airlock, or other access facility which requires passage through two sets of doors before entering the animal room.
• The interior surfaces of walls, floors, and ceilings are water resistant so that they may be easily cleaned. Penetrations in these surfaces are sealed or capable of being sealed to facilitate fumigation or space decontamination.
• A foot, elbow, or automatically operated hand washing sink is provided in each animal room near the exit door.
• If vacuum service (i.e., central or local) is provided, each service connection should be fitted with liquid disinfectant traps and a HEPA filter.
• If floor drains are provided, they are protected with liquid traps that are always filled with water or disinfectant.
• Windows in the animal room are non-operating and sealed.
• Animal room doors are self-closing and are kept closed when infected animals are present.
• An autoclave for decontaminating wastes is available, preferably within the animal facility. Materials are transferred to the autoclave in a covered leakproof container whose outer surface has been decontaminated.
• A non-recirculating ventilation system is provided. The supply and exhaust components of the system are balanced to provide for directional flow of air into the animal room. The exhaust air is discharged directly to the outside and clear of occupied areas and air intakes. Exhaust air from the room can be discharged without filtration or other treatment. Personnel must periodically validate that proper directional airflow is maintained.
• The HEPA filtered exhaust air from Class I or Class II biological safety cabinets or other primary containment devices is discharged directly to the outside or through the building exhaust system. Exhaust air from these primary containment devices may be recirculated within the animal room if the device is tested and certified at least every 12 months. If the HEPA filtered exhaust air from Class I or Class II biological safety cabinets is discharged to the outside through the building exhaust system, it is connected to this system in a manner (e.g., thimble unit connection) that avoids any interference with the performance of either the cabinet or building exhaust system.