How to Select a Suitable Biosafety Cabinet for Your Laboratory

By Wimala Horwongsakul

How Biosafety Level is Defined?

Infective Microorganisms by Risk Group

Definition Risk Group 1 (no or low individual and community risk)
A microorganism that is unlikely to cause human or animal disease

Examples
E. coli K12, Asporogenic Bacillus subtilis Baculoviruses, Herpesvirus saimiri

Risk Group 2 (moderate individual risk, low community risk)

A pathogen that can cause human or animal disease but is unlikely to be a serious hazard to laboratory workers, the community, livestock or the environment. Laboratory exposures may cause serious infection, but effective treatment and preventive measures are available and the risk of spread of infection is limited

E. coli, Neisseria meningitidis Treponema pallidum Cryptococcus neoformans Ascaris spp., Leishmania spp. Adenovirus. Hepatitis A E

Risk 3 (high individual risk, low community risk)

A pathogen that usually causes serious human or animal disease but does not ordinarily spread from one infected individual to another. Effective and preventive measures are available

Brucella spp. Coxsiella burnetti Mycobacterium tuberculosis Coccidioides immitis Alphaviruses (VEE) Hantaviruses, Monkeypox
Lassa virus Machupo virus Ebola virus Marburg virus Herpesvirus simiae Hemorrhagic fever viruses

Risk 4 (high individual and community risk)

A pathogen that usually causes serious human or animal disease and that can be readily transmitted from one individual to another, directly or indirectly. Effective treatment and preventive measures are not usually available.

Source: WHO Laboratory Biosafety Manual. 3rd Edition. 2004

Relation of Risk Groups to Biosafety Levels

Source: WHO Laboratory Biosafety Manual. 3rd Edition. 2004

Safety Biosafety
Biosafety Level 1
(BSL1)

. . .work is done with defined and characterized strains of viable microorganisms not known to consistently cause disease in healthy adult humans.
. . . work is done with the broad spectrum of indigenous moderate-risk agents that are present in the community and associated with human disease of varying severity.

. . . standard microbiological practices with no special primary or secondary barriers recommended, other than a sink for handwashing.
. . . procedures with aerosol or high splash potential that may increase the risk of such personnel exposure must be conducted in primary containment equipment, or in devices such as a BIOLOGICAL SAFETY CABINET or safety centrifuge cups. All laboratory manipulations should be performed in a BIOLOGICAL SAFETY CABINET or other enclosed equipment, such as a gas-tight aerosol generation chamber. All manipulations of infectious materials within the facility must be conducted in the Class III biological safety cabinet OR All procedures must be conducted by personnel wearing a one-piece positive pressure suit ventilated with a life support system.

Biosafety Level 2
(BSL2)

Biosafety Level 3
(BSL3)

. . .work is done with indigenous or exotic agents with a potential for respiratory transmission, and which may cause serious and potentially lethal infection.

Biosafety Level 4
(BSL4)

. . . work with dangerous and exotic agents that pose a high individual risk of lifethreatening disease, which may be transmitted via the aerosol route and for which there is no available vaccine or therapy.

Source: NSF International Standard for Biosafety Cabinetry, 2004

Summary of Biosafety Level Requirements

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Source: WHO Laboratory Biosafety Manual. 3rd Edition. 2004

A Typical Biosafety Level 1 Laboratory

Risk Group Biosafety Level Laboratory Type Laboratory Practices Safety Equipment

1 Basic Biosafety Level 1 Basic Teaching, Research GMT None; open bench work

GMT = good microbiological techniques

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A Typical Biosafety Level 2 Laboratory

Risk Group
Biosafety Level Laboratory Type

2
Basic Biosafety Level 2 Primary health services; diagnostic services, research GMT plus protective clothing, biohazard signs Open bench plus BSC for aerosols

Laboratory Practices

Safety Equipment

Procedures likely to generate aerosols are performed within a biological safety cabinet. Doors are kept closed and are posted with appropriate hazard signs. Potentially contaminated wastes are separated from the general waste stream
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A Typical Biosafety Level 3 Laboratory

Risk Group Biosafety Level

Laboratory Type Laboratory Practices

3 Basic Biosafety Level 3

Special diagnostic services, research Level 2 + special clothing, access control, directed airflow BSC and / or primary devices for all activities

Safety Equipment

The laboratory is accessed through an anteroom (double door entry or basic laboratory Biosafety Level 2) or an airlock. An autoclave is available within the facility for decontamination of wastes prior to disposal. A sink with hands-free operation is available. Inward directional airflow is established and all work with infectious materials is conducted within a biological safety cabinet.
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Classification of BSC How should we Select BSC?

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Types of BSC

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Differences between Class I, II and III BSCs

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Class I BSC
For personnel and environmental protection but not for product protection, having an open front, with negative-pressure inward airflow for personnel protection, and HEPA filtered exhaust air for environmental protection. Use to enclose equipment (e.g. centrifuges, small fermenter) or for procedure which may generate aerosols (e.g. tissue homogenization)

Does not have to be ducted, but the vast 0.38 m/s majority are ducted. Can be used in BSL3 labs

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Class III BSC

A totally enclosed, ventilated cabinet of leak-tight construction. Operations in the cabinet are conducted through tightly attached gloves. The cabinet is maintained under negative air pressure of at least 120 Pa below that of the experiment. Downflow air is drawn into the cabinet through HEPA filters. The exhaust air is treated by double HEPA filtration or by HEPA filtration and subsequent incineration.

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Class II BSC
For personnel, product and environmental protection, having an open front, via which the operator can operate in the cabinet, with negative-pressure inward airflow for personnel protection, downward HEPA filtered airflow for product protection and HEPA filtered exhausted air for environmental protection. Can be used with infectious agents in Risk Group 2 & 3.

Can be used with Risk Group 4 agents when positive-pressure suits are used

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The Types of Class II BSC

Four Key Elements

Comparison of Types
Class II, Type A1 BSC Class II, Type A2 BSC

BSC Exhaust
To Room or through canopy To Room or through canopy

Minimum Inflow

Exposed Positive Pressure Contaminated Plenums

Recirculation

0.4 m/s 0.5 m/s 0.5 m/s 0.5 m/s

Allowed

Mixed

Not allowed

Mixed

Class II, Type B1 BSC

Class II, Type B2 BSC

Direct Connection

Not allowed

Partial

Direct Connection

Not allowed

None

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Types of Class II BSC

30%
Total Exhaust
0.51m/s
0.51m/s 0.38m/s 0.51m/s

70%

Contaminated plenum under +ve pressure Min. intake 0.38m/s Not for volatile toxic chemicals

Contaminated ducts and plenum under -ve pressure Min. intake 0.51m/s Can be for minute quantity of volatile hazardous material if properly exhausted

Contaminated ducts and plenum under -ve pressure Min. intake 0.51m/s Can be for minute quantity of volatile hazardous material

Contaminated ducts and plenum under -ve pressure Min. intake 0.51m/s Reserved for works where volatile material recirculation must be avoided

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Class II Type A2 BSC

Must be externally exhausted Minimum inflow of 0.5 m/s No positively pressurized contaminated plenums Mixed recirculation

0.5 m/s

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Class II Type B2 BSC

Must be externally exhausted Minimum inflow of 100 fpm / 0.51 m/s No positively pressurized contaminated plenums No recirculation / Total Exhaust

No partial recirculation No mixed recirculation

0.5 m/s

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B2 Better than A2?

B2 are designed for applications involving volatile toxic chemicals or radionuclides (in addition to microbiological work) B2 do not provide a higher degree of biological protection B2 requires direct duct with dedicated fans. A2 can be configured as re-circulating. They can also be Ducted and used as Thimbles if there are traces of chemicals present B2 is exhausting nearly 2 times more air directly out of the Laboratory 494 m3/h A2 30% Exhausted 1220 m3/h

740 m3/h

B2 100% Externally Exhausted

Extra Cost External Motor / Fan Ductwork / Damper Volume of air exhausted Electricity consumption

494

m3/h

808 m3/h

480 m3/h

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Selection of BSC by Risk Group

Type of Protection Personnel protection, microorganisms in Risk Group 1-3 Personnel protection, microorganisms in Risk Group 4, glove-box laboratory BSC Selection Class I, Class II, Class III Class III Class I, Class II Class II, Class III only if laminar flow included

Personnel protection, microorganisms in Risk Group 4 suit laboratory

Product Protection

Volatile radionuclide / chemical protection, minute amounts

Volatile radionuclide / chemical protection

Class II B1, Class II A2 vented to the outside

Class I, Class II B2, Class III

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Source: WHO Laboratory Biosafety Manual. 3rd Edition. 2004

Selection of BSC by Risk Assessment

Source: CDC and NIH: Primary Containment of Biohazards: Selection, Installation and Use of BSC. 2 nd Edition. 2000

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We Should Care for Inflow Air

Air barrier at the front opening is susceptible to air movement
Located out of traffic patterns and away from room air currents.

Other devices that might move air

Room supply air vent Ventilation fans from electronic devices Swinging room doors Doors from refrigerators, autoclaves, and incubators Standard room fans or window-mounted air conditioners
Locations [1], [2], [3] are not suitable because they are exposed to drafts from windows and doors Location [5] is undesirable because it is in range of plant traffic and within the exhaust air range of a ventilation system [4] Preferred locations [6], [7], [8] because they are in a draftfree section of the room and not exposed to plant traffic
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