ANIMAL BIOSAFETY

PART 2
RECOMMENDED BIOSAFETY LEVEL
FOR INFECTIOUS ANIMALS

Summary of Recommended Biosafety

Levels for Activities in which
experimentally or naturally infected
vertebrate animals
 RECOMMENDED BIOSAFETY LEVEL FOR INFECTIOUS ANIMALS

ANIMAL BIOSAFETY LEVEL

• Animal facilities, like laboratories, may be designated With respect to animals to be used in the animal
according to a risk assessment and the risk group of the laboratory, factors for consideration include:
microorganisms under investigation, as Animal facility 1. The nature of the animals, i.e. their aggressiveness
Biosafety Level 1, 2, 3 or 4. and tendency to bite and scratch.
• With respect to agents to be used in the animal laboratory, 2. Their natural ecto- and endoparasites
factors for consideration include: 3. The zoonotic diseases to which they are susceptible.
4. The possible dissemination of allergens.
1. The normal route of transmission.
2. The volumes and concentrations to be used. As with laboratories, the requirements for design features,
3. The route of inoculation. equipment and precautions increase in stringency
4. Whether and by what route these agents may be excreted. according to the animal biosafety level.
 RECOMMENDED BIOSAFETY LEVEL FOR INFECTIOUS ANIMALS

ANIMAL BIOSAFETY LEVEL 1

This is suitable for the maintenance of most stock animals after quarantine (except nonhuman primates, regarding which
national authorities should be consulted), and for animals that are deliberately inoculated with agents in Risk Group 1. GMT
are required. The animal facility director must establish policies, procedures and protocols for all operations, and for access
to the vivarium. An appropriate medical surveillance programme for the staff must be instituted. A safety or operations
manual must be prepared and adopted.

Agents: Practices :
Not known to consistently Standard animal care and
cause diseases in healthy management practices, including
human adults. appropriate medical surveillance
programs.

Safety Equipment(Primary Facilities (Secondary Barriers):

Barriers): Standard animal facility:
As required for normal care • Non recirculation of exhaust air
of each species. • directional air flow
recommended.
• Hand washing sink is available
 RECOMMENDED BIOSAFETY LEVEL FOR INFECTIOUS ANIMALS

ANIMAL BIOSAFETY LEVEL 2

This is suitable for work with animals that are deliberately inoculated with microorganisms in Risk Group 2.
The following safety precautions apply: 11. Work space should be decontaminated after work.
1. All the requirements for animal facilities – Biosafety Level 1 12. Biological safety cabinets (Classes I or II) or isolator
must be met. cages with dedicated air supplies and HEPA-filtered
2. Biohazard warning signs should be posted on doors and other exhaust air must be provided for work that may involve
appropriate places. the generation of aerosols.
3. The facility must be designed for easy cleaning and 13. An autoclave must be available on site or in
housekeeping. appropriate proximity to the animal facility.
4. Doors must open inwards and be self-closing. 14. Animal bedding materials must be removed in a
5. Heating, ventilation and lighting must be adequate. manner that minimizes the generation of aerosols and
6. If mechanical ventilation is provided, the airflow must be dust.
inwards. Exhaust air is discharged to the outside and should not 15. All waste materials and bedding must be
be recirculated to any part of the building. decontaminated before disposal.
7. Access must be restricted to authorized persons. 16. Use of sharp instruments should be restricted
8. No animals should be admitted other than those for whenever possible. Sharps should always be collected in
experimental use. puncture-proof/-resistant containers fitted with covers
9. There should be an arthropod and rodent control programme. and treated as infectious.
10. Windows, if present, must be secure, resistant to breakage 17. Material for autoclaving or incineration must be
and, if able to be opened, must be fitted with arthropod-proof transported safely, in closed containers.
screens. 18. Animal cages must be decontaminated after use.
19. Animal carcasses should be incinerated.
20. Protective clothing and equipment must be worn in the facility, and removed on leaving.
21. Hand-washing facilities must be provided. Staff must wash their hands before leaving the animal facility.
22. All injuries, however minor, must be treated appropriately, reported and recorded.
23. Eating, drinking, smoking and application of cosmetics must be forbidden in the facility.
24. All personnel must receive appropriate training.

Agents: Practices :
Associated with human • ABSL - 1 practices plus: limited
disease. access.
Hazard:- percutaneous • biohazard warning signs.
exposure, ingestion, mucous • sharps precautions; biosafety
membrane. manual.
• Decontamination of all infectious
Safety Equipment(Primary
wastes and of animal cages prior to
Barriers):
washing.
• ABSL - I equipment plus
primary barriers: Facilities (Secondary Barriers):
containment equipment • ABSL - 1 facility plus.
appropriate for animal • Autoclave available.
species; • Handwashing sink available in the
• PPEs: laboratory coats animal room.
gloves, face and respiratory • Mechanical cage washer
protection as needed. recommended.
 RECOMMENDED BIOSAFETY LEVEL FOR INFECTIOUS ANIMALS

ANIMAL BIOSAFETY LEVEL 3

This is suitable for work with animals that are deliberately inoculated with agents in Risk Group 3, or when otherwise indicated
by a risk assessment. All systems, practices and procedures need to be reviewed and recertified annually.
The following safety precautions apply:
1. All the requirements for animal facilities – Biosafety Levels 1 and 2 must be
met.
2. Access must be strictly controlled.
3. The facility must be separated from other laboratory and animal house areas
by a room with a double-door entrance forming an anteroom.
4. Hand-washing facilities must be provided in the anteroom.
5. Showers should be provided in the anteroom.
6. There must be mechanical ventilation to ensure a continuous airflow through
all the rooms. Exhaust air must pass through HEPA filters before being
discharged to the atmosphere without recirculation. The system must be
designed to prevent accidental reverse flow and positive pressurization in any
part of the animal house.
7. An autoclave must be available at a location convenient for the animal house
where the biohazard is contained. Infectious waste should be autoclaved
before it is moved to other areas of the facility.
8. An incinerator should be readily available on site or alternative arrangements
should be made with the authorities concerned.
 RECOMMENDED BIOSAFETY LEVEL FOR INFECTIOUS ANIMALS

ANIMAL BIOSAFETY LEVEL 3

9. Animals infected with Risk Group 3 microorganisms must be housed in cages in isolators or rooms with ventilation exhausts
placed behind the cages.
10. Bedding should be as dust-free as possible.
11. All protective clothing must be decontaminated before it is laundered.
12. Windows must be closed and sealed, and resistant to breakage.
13. Immunization of staff, as appropriate, should be offered.

Agents:
Indigenous or exotic agents with Safety Equipment(Primary Barriers): Facilities (Secondary Barriers):
potential for aerosol transmission;
disease may have serious health • ABSL -2, equipment plus. • ABSL -2, facility plus.
effects. • Containment equipment for • physical separation from access
housinganimals and cage dumping corridors.
Practices : activities • self-closing, double-door
• ABSI -2 practices plus: controlled • Class I or II or II BSCs available for access.
access; decontamination of manipulative procedures • sealed penetrations.
clothing before laundering (inoculation, necropsy) that may • sealed windows autoclave
• cages decontaminated before create infectious aerosols. available in facility.
bedding removed • PPEs: Appropriate respiratory
• disinfectant foot bath as needed. protection.
 RECOMMENDED BIOSAFETY LEVEL FOR INFECTIOUS ANIMALS

ANIMAL BIOSAFETY LEVEL 4

Work in this facility will normally be linked with that in the maximum containment laboratory – Biosafety Level 4, and
national and local rules and regulations must be harmonized to apply to both. If work is to be done in a suit laboratory,
additional practices and procedures must be used over.
1. All the requirements for animal facilities – Biosafety Levels 1, 2 and 3
must be met.
2. Access must be strictly controlled; only staff designated by the
director of the establishment should have authority to enter.
3. Individuals must not work alone: the two-person rule must apply.
4. Personnel must have received the highest possible level of training
as microbiologists and be familiar with the hazards involved in their
work and with the necessary precautions.
5. Housing areas for animals infected with Risk Group 4 agents must
maintain the criteria for containment described and applied for
maximum containment laboratories – Biosafety Level 4.
6. The facility must be entered by an airlock anteroom, the clean side
of which must be separated from the restricted side by changing and
showering facilities.
7. Staff must remove street clothing when entering and put on special,
protective clothing. After work they must remove the protective
clothing for autoclaving, and shower before leaving.
8. A pass-through airlock with the clean end in a room outside the containment rooms must be provided for exchange of non-
autoclavable materials.
9. All manipulations with animals infected with Risk Group 4 agents must take place under maximum containment – Biosafety
Level 4 conditions.
10. All animals must be housed in isolators.
11. All animal bedding and waste must be autoclaved before removal from the facility.
12. There must be medical supervision of staff. The facility must be ventilated by a HEPA-filtered exhaust system designed to
ensure a negative pressure (inward directional airflow).
13. The ventilation system must be designed to prevent reverse flow and positive pressurization.
14. A double-ended autoclave with the clean end in a room outside the containment rooms must be provided for exchange of
materials.

Agents: Practices :
Dangerous/exotic agents that pose high Safety Equipment(Primary Barriers):
risk of life threatening disease; aerosol • ABSL - 3 practices plus
transmission, or related agents with • ABSL- 3 equipment plus
• entrance through change room
unknown risk of transmission. • Maximum containment
where personal clothing is
equipment (i.e., Class III BSC or
Facilities (Secondary Barriers): removed and laboratory
partial containment equipment in
• ABSL -3 facility plus clothing is put on
combination with full body, air-
• separate building or isolated zone • shower on exiting
supplied positive-pressure
• dedicated supply and exhaust, • all wastes are decontaminated
personnel suit) used for all
vacuum and decontamination before removal from the
procedures and activities.
systems. facility.
 Guidelines for animal laboratory / facility certification WHO Biosafety programme

In order to assure that adaptation and maintenance are undertaken promptly and in an appropriate and safe manner, all
biological research and clinical laboratories should be regularly certified.
Laboratory certification helps to ensure that:
1. Proper engineering controls are being used and are functioning adequately as designed
2. Appropriate site and protocol specific administrative controls are in place
3. Personal protective equipment is appropriate for the tasks being performed
4. Decontamination of waste and materials has been adequately considered and proper waste management procedures are
in place
5. Proper procedures for general laboratory safety, including physical, electrical and chemical safety are in place.
Laboratory certification differs from laboratory commissioning activities in several important ways. Laboratory certification is
the systematic examination of all safety features and processes within the laboratory (engineering controls, personal protective
equipment and administrative controls). Biosafety practices and procedures are also examined.

Laboratory certification is an on-going quality and safety assurance Certification of the laboratory should not be
activity that should take place on a regular basis. Adequately trained completed, and the laboratory should not be
safety and health or biosafety professionals may conduct laboratory declared functional, until deficiencies have been
certification activities. Institutions may employ personnel having the adequately addressed.
appropriate skill-set required for conducting audits, surveys or
inspections (these terms are used interchangeably) associated with
the certification process. However, institutions may consider engaging The complexity of Biosafety Level 4 laboratory
or be required to engage a third party to provide these services. operations goes beyond the scope of this manual.
 SUMMARY

INFECTION BIOSAFETY BIOSAFETY LEVEL FOR BIOSAFETY LEVEL FOR

INFECTIOUS AGENTS INFECTIOUS ANIMALS
CAUSES OF INFECTION PRIMARY CONTAINMENT BIOSAFETY LEVEL 1 ANIMAL BIOSAFETY LEVEL 1
TRANSMISSION
SECONDARY CONTAINMENT BIOSAFETY LEVEL 2 ANIMAL BIOSAFETY LEVEL 2
IDENTIFICATION OF
INFECTION ELEMENTS OF CONTAINMENT BIOSAFETY LEVEL 3 ANIMAL BIOSAFETY LEVEL 3

PREVENTING INFECTION THE LEVEL OF CONTAINMENT BIOSAFETY LEVEL 4 ANIMAL BIOSAFETY LEVEL 4

Guidelines for laboratory / Guidelines for laboratory /

facility certification facility certification
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