biohazrd a biological agent, such as an infectious microorganism, that constitutes a threat to...

Biohazard, Biological Biohazard, Biological Containment and Containment and

Biosafety CabinetsBiosafety Cabinets

Biohazard, Biological Biohazard, Biological Containment and Containment and

Biosafety CabinetsBiosafety Cabinets

Dr. Devendra Kumar Dr. Devendra Kumar NiranjanNiranjan

29-08-200829-08-2008

Dr. Devendra Kumar Dr. Devendra Kumar NiranjanNiranjan

29-08-200829-08-2008

BIOHAZRD

A biological agent, such as an A biological agent, such as an infectious microorganism, that infectious microorganism, that constitutes a threat to humans constitutes a threat to humans or to the environment, or to the environment, especially one produced in especially one produced in biological research or biological research or experimentation.experimentation.

A biological agent, such as an A biological agent, such as an infectious microorganism, that infectious microorganism, that constitutes a threat to humans constitutes a threat to humans or to the environment, or to the environment, especially one produced in especially one produced in biological research or biological research or experimentation.experimentation.

CLASSIFICATION

Risk Group 1Risk Group 1


Risk Group 3

Risk Group 4



Risk Group 3

Risk Group 4

No or low individual No or low individual and community riskand community risk

Moderate individual Moderate individual risk, low community risk, low community risksrisks

High individual risk, low community risk

High individual and community risk

No or low individual No or low individual and community riskand community risk

Moderate individual Moderate individual risk, low community risk, low community risksrisks

High individual risk, low community risk

High individual and community risk

Risk Group 1 No or low individual and

community risk

Risk Group 1 No or low individual and

community risk A microorganism that is

unlikely to cause disease in healthy human or animals

A microorganism that is unlikely to cause disease in healthy human or animals

Risk Group 2 Moderate individual risk, low

community risk

Risk Group 2 Moderate individual risk, low

community risk Risk Group 2 infectious agents are Risk Group 2 infectious agents are

pathogens that can cause human or pathogens that can cause human or animal disease but, under normal animal disease but, under normal circumstances, are unlikely to be a circumstances, are unlikely to be a serious hazard to laboratory serious hazard to laboratory workers, the community, or the workers, the community, or the environmentenvironment

Effective treatment and preventive Effective treatment and preventive measures are available and the risk measures are available and the risk of spread is limited.of spread is limited.

Risk Group 2 infectious agents are Risk Group 2 infectious agents are pathogens that can cause human or pathogens that can cause human or animal disease but, under normal animal disease but, under normal circumstances, are unlikely to be a circumstances, are unlikely to be a serious hazard to laboratory serious hazard to laboratory workers, the community, or the workers, the community, or the environmentenvironment

Effective treatment and preventive Effective treatment and preventive measures are available and the risk measures are available and the risk of spread is limited.of spread is limited.

Risk Group 2(contd.)

Risk Group 2 infectious agents Risk Group 2 infectious agents include : include : E. coli; salmonella; E. coli; salmonella; some some fungi fungi like ringworm; like ringworm; Viruses likeViruses like Encephalitis Encephalitis viruses; Human herpes simplex viruses; Human herpes simplex viruses; many influenza viruses; many influenza viruses; Mouse Hepatitis Virus; viruses; Mouse Hepatitis Virus; and a few parasitesand a few parasites..

Risk Group 3High individual risk, low

community risk

Risk Group 3High individual risk, low

community risk

Risk Group 3 infectious agents Risk Group 3 infectious agents are pathogens that usually are pathogens that usually cause serious human or animal cause serious human or animal disease, but do not ordinarily disease, but do not ordinarily spread by casual contact from spread by casual contact from one individual to another or that one individual to another or that can be treated by antimicrobial can be treated by antimicrobial or antiparasitic agents. or antiparasitic agents.

Risk Group 3 infectious agents Risk Group 3 infectious agents are pathogens that usually are pathogens that usually cause serious human or animal cause serious human or animal disease, but do not ordinarily disease, but do not ordinarily spread by casual contact from spread by casual contact from one individual to another or that one individual to another or that can be treated by antimicrobial can be treated by antimicrobial or antiparasitic agents. or antiparasitic agents.


Risk Group 3 pathogens include Risk Group 3 pathogens include BACTERIAS BACTERIAS such as such as Bacilluus, Bacilluus,

Brucella, Chlamydia, M. Brucella, Chlamydia, M. tuberculosis, Rickettsia, Salmonella tuberculosis, Rickettsia, Salmonella typhi, Shigella dysentriae type 1, typhi, Shigella dysentriae type 1, Yersinia pestisYersinia pestis etc.etc.

FUNGI FUNGI such as such as Histoplasma, Histoplasma, BlastomycesBlastomyces, etc., etc.

VIRUSES VIRUSES such as such as Hepatitis Hepatitis A,B,C,D,E virus, Herpes virus, HIV, A,B,C,D,E virus, Herpes virus, HIV, HTLV, Rabies virusHTLV, Rabies virus etc.etc.

Risk Group 4 High individual risk, high

community risk Risk Group 4 infectious agents are Risk Group 4 infectious agents are

pathogens that usually produce pathogens that usually produce very serious human or animal very serious human or animal disease, often disease, often untreatableuntreatable, and , and may be readily transmitted from may be readily transmitted from one individual to another, or from one individual to another, or from animal to human or vice-versa animal to human or vice-versa directly or indirectly, or by casual directly or indirectly, or by casual contact.contact.

Risk Group 4 infectious agents are Risk Group 4 infectious agents are pathogens that usually produce pathogens that usually produce very serious human or animal very serious human or animal disease, often disease, often untreatableuntreatable, and , and may be readily transmitted from may be readily transmitted from one individual to another, or from one individual to another, or from animal to human or vice-versa animal to human or vice-versa directly or indirectly, or by casual directly or indirectly, or by casual contact.contact.


Risk Group 4 infectious agents Risk Group 4 infectious agents are all viruses, such as,are all viruses, such as, Ebola Ebola viruses, Herpes B virus, Variola viruses, Herpes B virus, Variola virus, prions.virus, prions.

BIOLOGICAL CONTAINMENTBIOLOGICAL

CONTAINMENT Any methods to contain Any methods to contain

organisms by creating organisms by creating biochemical barriers to biochemical barriers to prevent them from growing prevent them from growing outside the laboratory. Oroutside the laboratory. Or

system designed to prevent system designed to prevent the accidental release of the accidental release of biological material.biological material.

Any methods to contain Any methods to contain organisms by creating organisms by creating biochemical barriers to biochemical barriers to prevent them from growing prevent them from growing outside the laboratory. Oroutside the laboratory. Or

system designed to prevent system designed to prevent the accidental release of the accidental release of biological material.biological material.

Biosafety Level 1 (Containment Level 1)

No special facilities, equipment or No special facilities, equipment or procedures are requiredprocedures are required

Hand-washing facilities must be provided.Hand-washing facilities must be provided. Disinfectants must be properly used.Disinfectants must be properly used. Protective laboratory clothing that has been

used in the laboratory must not be stored in the same lockers or cupboards as street clothing.

Appropriate gloves must be worn for all procedures that may involve direct or accidental contact with potentially infectious materials

Open-toed footwear must not be worn in laboratories.

No special facilities, equipment or No special facilities, equipment or procedures are requiredprocedures are required

Hand-washing facilities must be provided.Hand-washing facilities must be provided. Disinfectants must be properly used.Disinfectants must be properly used. Protective laboratory clothing that has been

used in the laboratory must not be stored in the same lockers or cupboards as street clothing.

Appropriate gloves must be worn for all procedures that may involve direct or accidental contact with potentially infectious materials

Open-toed footwear must not be worn in laboratories.

Biosafety Level 1 Eating, drinking, smoking, applying

cosmetics and handling contact lenses is prohibited in the laboratory working areas.

Storing human foods or drinks anywhere in the laboratory working areas is prohibited.

Pipetting by mouth must be strictly forbidden.

Materials must not be placed in the mouth. Labels must not be licked.

The use of hypodermic needles and syringes should be limited.

Laboratory coveralls, gowns or uniforms must be worn at all times for work in the laboratory.

Eating, drinking, smoking, applying cosmetics and handling contact lenses is prohibited in the laboratory working areas.

Storing human foods or drinks anywhere in the laboratory working areas is prohibited.

Pipetting by mouth must be strictly forbidden.

Materials must not be placed in the mouth. Labels must not be licked.

The use of hypodermic needles and syringes should be limited.

Laboratory coveralls, gowns or uniforms must be worn at all times for work in the laboratory.

Biosafety Level 1Lab. Ample space must be provided for the safe

conduct of laboratory work and for cleaning and maintenance.

Walls, ceilings and floors should be smooth, easy to clean, impermeable to liquids and resistant to the chemicals and disinfectants normally used in the laboratory.

Floors should be slip-resistant. Bench tops should be impervious to water

and resistant to disinfectants, acids, alkalis, organic solvents and moderate heat.

Ample space must be provided for the safe conduct of laboratory work and for cleaning and maintenance.

Walls, ceilings and floors should be smooth, easy to clean, impermeable to liquids and resistant to the chemicals and disinfectants normally used in the laboratory.

Floors should be slip-resistant. Bench tops should be impervious to water

and resistant to disinfectants, acids, alkalis, organic solvents and moderate heat.

Biosafety Level 1Lab. Illumination should be adequate for all

activities. Open spaces between and under benches,

cabinets and equipment should be accessible for cleaning.

Storage space must be adequate to hold supplies for immediate use and thus prevent clutter on bench tops and in aisles.

Additional long-term storage space, conveniently

located outside the laboratory working areas, should also be provided.

Illumination should be adequate for all activities.

Open spaces between and under benches, cabinets and equipment should be accessible for cleaning.

Storage space must be adequate to hold supplies for immediate use and thus prevent clutter on bench tops and in aisles.

Additional long-term storage space, conveniently

located outside the laboratory working areas, should also be provided.

Biosafety level 1 Lab.

Hand-washing basins, with running water if possible, should be provided in each laboratory room, preferably near the exit door.

Doors should have vision panels, appropriate fire ratings, and preferably be self closing.

A dependable supply of good quality water is essential.

Hand-washing basins, with running water if possible, should be provided in each laboratory room, preferably near the exit door.

Doors should have vision panels, appropriate fire ratings, and preferably be self closing.

A dependable supply of good quality water is essential.


There should be a reliable and adequate electricity supply and emergency lighting to permit safe exit.

There should be a reliable and adequate supply of gas

The laboratory should be kept neat, & clean

Work surfaces must be decontaminated after any spill of potentially dangerous material and at the end of the working day

There should be a reliable and adequate electricity supply and emergency lighting to permit safe exit.

There should be a reliable and adequate supply of gas

The laboratory should be kept neat, & clean

Work surfaces must be decontaminated after any spill of potentially dangerous material and at the end of the working day

Biosafety level 2 (Containment Level 2)

The international biohazard warning symbol and sign must be displayed on the doors of the rooms where microorganisms of Risk Group 2 or higher risk groups are handled.

The international biohazard warning symbol and sign must be displayed on the doors of the rooms where microorganisms of Risk Group 2 or higher risk groups are handled.

BIOHAZARDADMITTANCE TO AUTHORIZED PERSONNEL ONLYBiosafety Level: _________________________________Responsible Investigator: _________________________In case of emergency call: ________________________Daytime phone: __________Home phone: ___________Authorization for entrance must be obtained from the

Responsible Investigator named above

BIOHAZARDADMITTANCE TO AUTHORIZED PERSONNEL ONLYBiosafety Level: _________________________________Responsible Investigator: _________________________In case of emergency call: ________________________Daytime phone: __________Home phone: ___________Authorization for entrance must be obtained from the

Responsible Investigator named above

Biosafety level 2 Only authorized persons should be

allowed to enter the laboratory working areas.

Laboratory doors should be kept closed.

Children should not be authorized or allowed to enter laboratory working areas.

Safety glasses, face shields (visors) or other protective devices must be worn when it is necessary

Only authorized persons should be allowed to enter the laboratory working areas.

Laboratory doors should be kept closed.

Children should not be authorized or allowed to enter laboratory working areas.

Safety glasses, face shields (visors) or other protective devices must be worn when it is necessary

Biosafety level 2 An autoclave or other means of

decontamination should be available in appropriate proximity to the laboratory.

Room surfaces should be impervious and readily cleanable.

Certified HEPA filtered class I or II Certified HEPA filtered class I or II biological safety cabinet for biological safety cabinet for organism manipulations.organism manipulations.

All contaminated material to be All contaminated material to be properly decontaminated properly decontaminated

An autoclave or other means of decontamination should be available in appropriate proximity to the laboratory.

Room surfaces should be impervious and readily cleanable.

Certified HEPA filtered class I or II Certified HEPA filtered class I or II biological safety cabinet for biological safety cabinet for organism manipulations.organism manipulations.

All contaminated material to be All contaminated material to be properly decontaminated properly decontaminated

Biosafety level 2 A preplacement health check is

necessary. The person’s medical history should be recorded and a targeted occupational health assessment performed.

Records of illness and absence should be kept by the laboratory management.

Women of childbearing age should be made aware of the risk to an unborn child of occupational exposure to certain microorganisms, e.g. rubella virus.

A preplacement health check is necessary. The person’s medical history should be recorded and a targeted occupational health assessment performed.

Records of illness and absence should be kept by the laboratory management.

Women of childbearing age should be made aware of the risk to an unborn child of occupational exposure to certain microorganisms, e.g. rubella virus.

Biosafety level 3 (Containment Level 3)

Laboratory protective clothing must be of the type with solid-front or wrap-around gowns, scrub suits, coveralls,& head covering.

Open manipulations of all potentially infectious material must be conducted within a biological safety cabinet or other primary containment device.

Respiratory protective equipment may be necessary.

Laboratory protective clothing must be of the type with solid-front or wrap-around gowns, scrub suits, coveralls,& head covering.

Open manipulations of all potentially infectious material must be conducted within a biological safety cabinet or other primary containment device.

Respiratory protective equipment may be necessary.

Biosafety level 3 The laboratory must be separated

from the areas that are open to unrestricted traffic flow within the building.

Lab should be with controlled access double door entry and body shower.

The laboratory room must be sealable for decontamination.

A hand-washing station with hands-free controls should be provided near each exit door.

The laboratory must be separated from the areas that are open to unrestricted traffic flow within the building.

Lab should be with controlled access double door entry and body shower.

The laboratory room must be sealable for decontamination.

A hand-washing station with hands-free controls should be provided near each exit door.

Biosafety level 3 An autoclave for the decontamination

of contaminated waste material should be available in the containment laboratory.

There must be a controlled ventilation system that maintains a directional airflow.

Ventilation system design must ensure that air pressure is negative to surrounding areas at all times, with no recirculation of air.

An autoclave for the decontamination of contaminated waste material should be available in the containment laboratory.

There must be a controlled ventilation system that maintains a directional airflow.

Ventilation system design must ensure that air pressure is negative to surrounding areas at all times, with no recirculation of air.

Biosafety level 3

Air exhausted through a Air exhausted through a dedicated exhaust or HEPA dedicated exhaust or HEPA filtration system filtration system

certified HEPA filtered class II certified HEPA filtered class II biological safety cabinet for biological safety cabinet for organism manipulations organism manipulations

Air exhausted through a Air exhausted through a dedicated exhaust or HEPA dedicated exhaust or HEPA filtration system filtration system

certified HEPA filtered class II certified HEPA filtered class II biological safety cabinet for biological safety cabinet for organism manipulations organism manipulations

Biosafety Level 4 (Containment Level 4)Biosafety Level 4

(Containment Level 4) Biosafety Level 4 is designed for

work with Risk Group 4 microorganisms.

The two-person rule should apply, whereby no individual ever works alone

A complete change of clothing and shoes is required prior to entering and upon exiting the laboratory.

Biosafety Level 4 is designed for work with Risk Group 4 microorganisms.

The two-person rule should apply, whereby no individual ever works alone

A complete change of clothing and shoes is required prior to entering and upon exiting the laboratory.

Biosafety Level 4 Primary containment Class III cabinet laboratory Suit laboratory (positive pressure ventilated ) ) Controlled access Controlled air system Decontamination of effluents Sterilization of waste and materials Airlock entry ports Emergency power Containment drain

Primary containment Class III cabinet laboratory Suit laboratory (positive pressure ventilated ) ) Controlled access Controlled air system Decontamination of effluents Sterilization of waste and materials Airlock entry ports Emergency power Containment drain

BIOSAFETY LEVEL 11 22 33 44Isolation of laboratory NONO NONO YEYE

SSYESYES

Room sealable for decontamination

NONO NONO YEYESS

YESYES

VentilationVentilationinward airflowinward airflow NONO DEDE YEYE

SSYESYES

controlled ventilating controlled ventilating systemsystem

NONO DEDE YEYESS

YESYES

HEPA-filtered air HEPA-filtered air exhaustexhaust

NONO NONO YES/YES/NONO

YESYES

Double-door entryDouble-door entry NONO NONO YEYESS

YESYES

AirlockAirlock NONO NONO NONO YESYES

BIOSAFETY REQUIREMENTS

Airlock with showerAirlock with shower NONO NONO NONO YESYESAnteroomAnteroom NONO NONO YEYE

SS------

Anteroom with showerAnteroom with shower NONO NONO NO/ NO/ YESYES

YESYES

Effluent treatmentEffluent treatment NONO NONO NO/ NO/ YESYES

YESYES

AutoclaveAutoclaveon siteon site NONO DEDE YEYE

SSYESYES

in laboratory roomin laboratory room NONO NONO DEDE YESYESdouble-endeddouble-ended NONO NONO DEDE YESYESBiological safety cabinetsBiological safety cabinets NONO DEDE YEYE

SSYESYES

Personnel safety Personnel safety monitoring capabilitymonitoring capability

NONO NONO DEDE YESYES

Biological Safety Cabinets (BSC)

Containment and protection devices Containment and protection devices used in laboratories working with used in laboratories working with biological agents with a primary biological agents with a primary purpose of protecting the laboratory purpose of protecting the laboratory worker and the environment from worker and the environment from viable organisms.viable organisms.

Class I Biological Safety Cabinets

Class I BSC is a ventilated Class I BSC is a ventilated cabinet in which rcabinet in which room air is drawn in through the front opening at a minimum velocity of 0.38 m/s, it passes over the work surface and is discharged from the cabinet through the exhaust duct . .

has a HEPA filter in the exhaust system to protect the environment

The Class I BSC provides personnel and environmental protection, but no product protection

Class I BSC is a ventilated Class I BSC is a ventilated cabinet in which rcabinet in which room air is drawn in through the front opening at a minimum velocity of 0.38 m/s, it passes over the work surface and is discharged from the cabinet through the exhaust duct . .

has a HEPA filter in the exhaust system to protect the environment

The Class I BSC provides personnel and environmental protection, but no product protection

Class I Biological Safety Cabinet

Class I Biological Safety Cabinets

Class II Biological Safety Cabinets

Class II BSC’s are Laminar Flow Biological Safety Class II BSC’s are Laminar Flow Biological Safety Cabinets that protect personnel, product, and Cabinets that protect personnel, product, and environment. environment.

They provide an They provide an inward airflowinward airflow to protect to protect personnel, a personnel, a downward flowdownward flow of HEPA filtered air of HEPA filtered air to the work area to protect the product, and to the work area to protect the product, and then then exhaust HEPA filtered airexhaust HEPA filtered air to protect the to protect the environment from particulate and aerosol environment from particulate and aerosol hazards.hazards.

Class II BSCs are partial barrier systems that rely on the laminar movement of air to provide containment

If the air curtain is disrupted (e.g., movement of materials in and out of a cabinet, rapid or sweeping movement of the arms) the potential for contaminant release into the laboratory work environment is increased

Class II BSC’s are Laminar Flow Biological Safety Class II BSC’s are Laminar Flow Biological Safety Cabinets that protect personnel, product, and Cabinets that protect personnel, product, and environment. environment.

They provide an They provide an inward airflowinward airflow to protect to protect personnel, a personnel, a downward flowdownward flow of HEPA filtered air of HEPA filtered air to the work area to protect the product, and to the work area to protect the product, and then then exhaust HEPA filtered airexhaust HEPA filtered air to protect the to protect the environment from particulate and aerosol environment from particulate and aerosol hazards.hazards.

Class II BSCs are partial barrier systems that rely on the laminar movement of air to provide containment

If the air curtain is disrupted (e.g., movement of materials in and out of a cabinet, rapid or sweeping movement of the arms) the potential for contaminant release into the laboratory work environment is increased

Class II Type A 1 BSC

Class || Type A1 BSC offer Class || Type A1 BSC offer personnel protection through personnel protection through negative pressure airflow into negative pressure airflow into the cabinet.the cabinet.

To protect the product, the To protect the product, the work area in the cabinet is work area in the cabinet is continuously bathed with continuously bathed with ultra-clean air provided by the ultra-clean air provided by the supply HEPA filter.supply HEPA filter.

Maintain a minimum average Maintain a minimum average face velocity of 0.38 m/s face velocity of 0.38 m/s

Approximately 70% of the air Approximately 70% of the air of each cycle is recirculated of each cycle is recirculated through this supply HEPA through this supply HEPA filter. filter.

The remaining air is The remaining air is discharged from the hood discharged from the hood through the exhaust HEPA through the exhaust HEPA filter, protecting the filter, protecting the environment.environment.

Exhaust to room or thimble connection vented to the outside

Class || Type A1 BSC offer Class || Type A1 BSC offer personnel protection through personnel protection through negative pressure airflow into negative pressure airflow into the cabinet.the cabinet.

To protect the product, the To protect the product, the work area in the cabinet is work area in the cabinet is continuously bathed with continuously bathed with ultra-clean air provided by the ultra-clean air provided by the supply HEPA filter.supply HEPA filter.

Maintain a minimum average Maintain a minimum average face velocity of 0.38 m/s face velocity of 0.38 m/s

Approximately 70% of the air Approximately 70% of the air of each cycle is recirculated of each cycle is recirculated through this supply HEPA through this supply HEPA filter. filter.

The remaining air is The remaining air is discharged from the hood discharged from the hood through the exhaust HEPA through the exhaust HEPA filter, protecting the filter, protecting the environment.environment.

Exhaust to room or thimble connection vented to the outside


A- Front OpeningA- Front OpeningB- SashB- SashC- Exhaust HEPA C- Exhaust HEPA

FilterFilterD- Rear PlenumD- Rear PlenumE- Supply HEPA E- Supply HEPA

FilterFilterF- BlowerF- Blower

Class II Type A 2 BSCClass II Type A 2 BSC

This biological safety cabinet is a ducted Type A cabinet having a minimum inward airflow of 0.51m/s

All positive pressure contaminated plenums within the cabinet are surrounded by a negative air pressure plenum.

leakage in a contaminated plenum will be into the cabinet and not into the environment

This biological safety cabinet is a ducted Type A cabinet having a minimum inward airflow of 0.51m/s

All positive pressure contaminated plenums within the cabinet are surrounded by a negative air pressure plenum.

leakage in a contaminated plenum will be into the cabinet and not into the environment


A. narrow front openingA. narrow front opening

B. sashB. sash

C. exhaust HEPA filterC. exhaust HEPA filter

D. supply HEPA filterD. supply HEPA filter

E. negative pressure exhaust E. negative pressure exhaust plenumplenum

F. supply blowerF. supply blower

A. narrow front openingA. narrow front opening

B. sashB. sash



E. negative pressure exhaust E. negative pressure exhaust plenumplenum

F. supply blowerF. supply blower

Class II Type B 1 BSCClass II Type B 1 BSC

An internal blower draws sufficient room air through the front grille to maintain a minimum average inflow velocity of at least 0.51m/s at the face opening of the cabinet.

Carcinogens used in Carcinogens used in cell culture or microbial cell culture or microbial systems require both systems require both biological and chemical biological and chemical containment.containment.

An internal blower draws sufficient room air through the front grille to maintain a minimum average inflow velocity of at least 0.51m/s at the face opening of the cabinet.

Carcinogens used in Carcinogens used in cell culture or microbial cell culture or microbial systems require both systems require both biological and chemical biological and chemical containment.containment.


The cabinet supply blowers draw room air through the front grille and a portion of the cabinet’s recirculated air through the supply HEPA filters located immediately below the work surface.

This particulate-free air flows upward through a plenum at each side of the cabinet and then downward to the work area through a back-pressure plate.

The cabinet supply blowers draw room air through the front grille and a portion of the cabinet’s recirculated air through the supply HEPA filters located immediately below the work surface.

This particulate-free air flows upward through a plenum at each side of the cabinet and then downward to the work area through a back-pressure plate.


Type B1 cabinets must be hard-ducted, preferably to a dedicated, independent exhaust system.

Approximately 70 percent of the down flow air exits through the rear grille, passes through the exhaust HEPA filter, and is discharged from the building. The remaining 30 percent of the down flow air is drawn through the front grille

Suitable for work with low levels of volatile Suitable for work with low levels of volatile toxic chemicals and trace amounts of toxic chemicals and trace amounts of radionuclides radionuclides

Type B1 cabinets must be hard-ducted, preferably to a dedicated, independent exhaust system.

Approximately 70 percent of the down flow air exits through the rear grille, passes through the exhaust HEPA filter, and is discharged from the building. The remaining 30 percent of the down flow air is drawn through the front grille

Suitable for work with low levels of volatile Suitable for work with low levels of volatile toxic chemicals and trace amounts of toxic chemicals and trace amounts of radionuclides radionuclides

Class II Type B 1 BSC Class II Type B 1 BSC

A- Front OpeningA- Front Opening

B- SashB- Sash

C- Exhaust HEPA FilterC- Exhaust HEPA Filter

D- Supply PlenumD- Supply Plenum

E- Supply HEPA FilterE- Supply HEPA Filter

F- BlowerF- Blower


This is a total-exhaust cabinet; no air is This is a total-exhaust cabinet; no air is recirculated within itrecirculated within it

This cabinet provides simultaneous primary This cabinet provides simultaneous primary biological and chemical containment.biological and chemical containment.

The supply blower draws in room air or The supply blower draws in room air or outside air at the top of the cabinet, passes outside air at the top of the cabinet, passes it through a HEPA filter and down into the it through a HEPA filter and down into the work area of the cabinetwork area of the cabinet..

Maintain a minimum average inflow velocity of at least 0.51m/s at the face opening of the cabinet

This is a total-exhaust cabinet; no air is This is a total-exhaust cabinet; no air is recirculated within itrecirculated within it

This cabinet provides simultaneous primary This cabinet provides simultaneous primary biological and chemical containment.biological and chemical containment.

The supply blower draws in room air or The supply blower draws in room air or outside air at the top of the cabinet, passes outside air at the top of the cabinet, passes it through a HEPA filter and down into the it through a HEPA filter and down into the work area of the cabinetwork area of the cabinet..

Maintain a minimum average inflow velocity of at least 0.51m/s at the face opening of the cabinet

Class II Type B 2 BSC

Hard-ducted through a dedicated duct exhausted to the atmosphere, 100% of cabinet air, after passage through a HEPA filter; contain negative pressure plenum.

This cabinet exhausts as much as 1200 cubic feet per minute of conditioned room air making this cabinet expensive to operate.

Exhaust air movement should be monitored by a pressure-independent device, such as a flow monitor.

Hard-ducted through a dedicated duct exhausted to the atmosphere, 100% of cabinet air, after passage through a HEPA filter; contain negative pressure plenum.

This cabinet exhausts as much as 1200 cubic feet per minute of conditioned room air making this cabinet expensive to operate.

Exhaust air movement should be monitored by a pressure-independent device, such as a flow monitor.

Class II Type B 2 BSC Schematic. A. front openingA. front opening

B. sashB. sash



E. negative pressure E. negative pressure exhaust plenumexhaust plenum

F. supply blower F. supply blower

G. filter screenG. filter screen

The cabinet exhaust The cabinet exhaust needs to be connected to needs to be connected to the building exhaust the building exhaust system.system.

Class II Type B 2 BSC

Class II BSC ModificationsClass II BSC Modifications

The front sash can be modified by the manufacturer The front sash can be modified by the manufacturer to accommodate a microscopeto accommodate a microscope

A rigid plate with arm holes can be added if needed. A rigid plate with arm holes can be added if needed. The work surface can be designed to accept a The work surface can be designed to accept a

centrifuge, or other equipment that requires centrifuge, or other equipment that requires containmentcontainment

Good cabinet design, microbiological aerosol tracer Good cabinet design, microbiological aerosol tracer testing of the modification, and appropriate testing of the modification, and appropriate certification are required to ensure that the basic certification are required to ensure that the basic BSC systems operate properly after modification.BSC systems operate properly after modification.

Maximum containment potential is achieved only Maximum containment potential is achieved only through strict adherence to proper practices and through strict adherence to proper practices and procedures.procedures.

The front sash can be modified by the manufacturer The front sash can be modified by the manufacturer to accommodate a microscopeto accommodate a microscope

A rigid plate with arm holes can be added if needed. A rigid plate with arm holes can be added if needed. The work surface can be designed to accept a The work surface can be designed to accept a

centrifuge, or other equipment that requires centrifuge, or other equipment that requires containmentcontainment

Good cabinet design, microbiological aerosol tracer Good cabinet design, microbiological aerosol tracer testing of the modification, and appropriate testing of the modification, and appropriate certification are required to ensure that the basic certification are required to ensure that the basic BSC systems operate properly after modification.BSC systems operate properly after modification.

Maximum containment potential is achieved only Maximum containment potential is achieved only through strict adherence to proper practices and through strict adherence to proper practices and procedures.procedures.

Class II BSCs can be modified to accommodate special tasks:

Class II BSCs can be modified to accommodate special tasks:

Class III BSC or Glovebox

The Class III BSC was designed for work with Biosafety Level 4 The Class III BSC was designed for work with Biosafety Level 4 microbiological agents and provides maximum protection to the microbiological agents and provides maximum protection to the environment and the worker.environment and the worker.

It is a gas-tight enclosure with a non-opening view window.It is a gas-tight enclosure with a non-opening view window. Access for passage of materials into the cabinet is through a dunk tank Access for passage of materials into the cabinet is through a dunk tank

that is accessible through the cabinet floor or double door pass-through that is accessible through the cabinet floor or double door pass-through box such as an autoclave that can be decontaminated between uses. box such as an autoclave that can be decontaminated between uses.

Reversing that process allows for safe removal of materials from the Class Reversing that process allows for safe removal of materials from the Class III BSC. III BSC.

Both supply and exhaust air are HEPA filtered. Exhaust air must pass Both supply and exhaust air are HEPA filtered. Exhaust air must pass through two HEPA filters or a HEPA filter and an air incinerator before through two HEPA filters or a HEPA filter and an air incinerator before discharge to the outdoors. discharge to the outdoors.

Airflow is maintained by a dedicated independent exhaust system exterior Airflow is maintained by a dedicated independent exhaust system exterior to the cabinet which keeps the cabinet under negative pressureto the cabinet which keeps the cabinet under negative pressure

The Class III BSC was designed for work with Biosafety Level 4 The Class III BSC was designed for work with Biosafety Level 4 microbiological agents and provides maximum protection to the microbiological agents and provides maximum protection to the environment and the worker.environment and the worker.

It is a gas-tight enclosure with a non-opening view window.It is a gas-tight enclosure with a non-opening view window. Access for passage of materials into the cabinet is through a dunk tank Access for passage of materials into the cabinet is through a dunk tank

that is accessible through the cabinet floor or double door pass-through that is accessible through the cabinet floor or double door pass-through box such as an autoclave that can be decontaminated between uses. box such as an autoclave that can be decontaminated between uses.

Reversing that process allows for safe removal of materials from the Class Reversing that process allows for safe removal of materials from the Class III BSC. III BSC.

Both supply and exhaust air are HEPA filtered. Exhaust air must pass Both supply and exhaust air are HEPA filtered. Exhaust air must pass through two HEPA filters or a HEPA filter and an air incinerator before through two HEPA filters or a HEPA filter and an air incinerator before discharge to the outdoors. discharge to the outdoors.

Airflow is maintained by a dedicated independent exhaust system exterior Airflow is maintained by a dedicated independent exhaust system exterior to the cabinet which keeps the cabinet under negative pressureto the cabinet which keeps the cabinet under negative pressure

Long, heavy-duty rubber gloves are attached in a gas-tight manner to ports in the cabinet and allow for manipulation of the materials isolated inside. Although these gloves restrict movement, they prevent the user's direct contact with the hazardous materials. Depending on the design of the cabinet, the supply HEPA filter provides particulate-free, albeit somewhat turbulent, airflow within the work environment.

Long, heavy-duty rubber gloves are attached in a gas-tight manner to ports in the cabinet and allow for manipulation of the materials isolated inside. Although these gloves restrict movement, they prevent the user's direct contact with the hazardous materials. Depending on the design of the cabinet, the supply HEPA filter provides particulate-free, albeit somewhat turbulent, airflow within the work environment.

Class III BSC or Glovebox

Class III BSC or Gloveboxes

Class III cabinets are usually only installed in maximum containment laboratories with controlled

access.

They require special ventilation or other support systems

such as steam for autoclaves.

Class III cabinets are usually only installed in maximum containment laboratories with controlled

access.

They require special ventilation or other support systems

such as steam for autoclaves.

Class III BSC or Glove boxes

Class III BSC Schematic

A- Glove PortsA- Glove Ports

B- SashB- Sash

C- Exhaust HEPA FilterC- Exhaust HEPA Filter

D- Supply HEPA FilterD- Supply HEPA Filter

E- Double Ended E- Double Ended Autoclave or Pass Autoclave or Pass Through BoxThrough Box

A chemical dunk tank may be installed beneath the work surface

of the BSC with access from above.

The cabinet exhaust needs to be connected to the building exhaust system.

A chemical dunk tank may be installed beneath the work surface

of the BSC with access from above.

The cabinet exhaust needs to be connected to the building exhaust system.

BSC FACE VELOCITY (m/s)

AIRFLOW (%)RECIRCUL-ATED

AIRFLOW (%)EXHAUST-ED

EXHAUST SYSTEM

Class I 0.36 00 100100 Hard duct

Class || A1

0.38–0.51 7070 3030 Exhaust to room or thimbleconnection

Class || A2vented to theoutside

0.51 70 3030 Exhaust to room or thimble connection

Class || B1

0.51 3030 70 Hard duct

Class || B2

0.51 00 100100 Hard duct

Class ||| NANA 00 100100 Hard duct

Start-up procedures Turn off UV lights if in use and ensure that the sash is Turn off UV lights if in use and ensure that the sash is

in the appropriate position.in the appropriate position. Turn on fluorescent light and cabinet blower, if off.Turn on fluorescent light and cabinet blower, if off. Check the air intake and exhaust grilles for Check the air intake and exhaust grilles for

obstructions.obstructions. If the cabinet is equipped with an alarm, test the alarm If the cabinet is equipped with an alarm, test the alarm

and switch it to the "on" position.and switch it to the "on" position. Confirm inward airflow by holding a tissue at the Confirm inward airflow by holding a tissue at the

middle of the edge of the viewing panel and ensuring middle of the edge of the viewing panel and ensuring that it is drawn in.that it is drawn in.

Disinfect the interior surfaces with a suitable, Disinfect the interior surfaces with a suitable, noncorrosive disinfectant.noncorrosive disinfectant.

Assemble all materials required for the procedure and Assemble all materials required for the procedure and load them into the cabinet; do not obstruct the air load them into the cabinet; do not obstruct the air grilles; grilles;

Wait 5 minutes to purge airborne contaminants from Wait 5 minutes to purge airborne contaminants from the work area.the work area.

Turn off UV lights if in use and ensure that the sash is Turn off UV lights if in use and ensure that the sash is in the appropriate position.in the appropriate position.

Turn on fluorescent light and cabinet blower, if off.Turn on fluorescent light and cabinet blower, if off. Check the air intake and exhaust grilles for Check the air intake and exhaust grilles for

obstructions.obstructions. If the cabinet is equipped with an alarm, test the alarm If the cabinet is equipped with an alarm, test the alarm

and switch it to the "on" position.and switch it to the "on" position. Confirm inward airflow by holding a tissue at the Confirm inward airflow by holding a tissue at the

middle of the edge of the viewing panel and ensuring middle of the edge of the viewing panel and ensuring that it is drawn in.that it is drawn in.

Disinfect the interior surfaces with a suitable, Disinfect the interior surfaces with a suitable, noncorrosive disinfectant.noncorrosive disinfectant.

Assemble all materials required for the procedure and Assemble all materials required for the procedure and load them into the cabinet; do not obstruct the air load them into the cabinet; do not obstruct the air grilles; grilles;

Wait 5 minutes to purge airborne contaminants from Wait 5 minutes to purge airborne contaminants from the work area.the work area.

Procedures upon completion of the work

Allow the cabinet to run for 5 minutes with no activity.Allow the cabinet to run for 5 minutes with no activity. Close or cover open containers before removing them Close or cover open containers before removing them

from the cabinet.from the cabinet. Surface disinfect objects in contact with contaminated Surface disinfect objects in contact with contaminated

material before removal from the cabinet.material before removal from the cabinet. Remove contaminated gloves and dispose of them as Remove contaminated gloves and dispose of them as

appropriate; wash hands.appropriate; wash hands. Using a suitable non-corrosive disinfectant (e.g., 70% Using a suitable non-corrosive disinfectant (e.g., 70%

ethanol), disinfect interior surfaces of cabinet; ethanol), disinfect interior surfaces of cabinet; periodically remove the work surface and disinfect the periodically remove the work surface and disinfect the area beneath it (including the catch pan) and wipe the area beneath it (including the catch pan) and wipe the surface of the UV light with disinfectant.surface of the UV light with disinfectant.

Turn off the fluorescent light and cabinet blower when Turn off the fluorescent light and cabinet blower when appropriate appropriate

Turn on the UV light if appropriate (do not turn on Turn on the UV light if appropriate (do not turn on when people are working close by); UV must be tested when people are working close by); UV must be tested to ensure that it is emitting a germicidal wavelength.to ensure that it is emitting a germicidal wavelength.

Allow the cabinet to run for 5 minutes with no activity.Allow the cabinet to run for 5 minutes with no activity. Close or cover open containers before removing them Close or cover open containers before removing them

from the cabinet.from the cabinet. Surface disinfect objects in contact with contaminated Surface disinfect objects in contact with contaminated

material before removal from the cabinet.material before removal from the cabinet. Remove contaminated gloves and dispose of them as Remove contaminated gloves and dispose of them as

appropriate; wash hands.appropriate; wash hands. Using a suitable non-corrosive disinfectant (e.g., 70% Using a suitable non-corrosive disinfectant (e.g., 70%

ethanol), disinfect interior surfaces of cabinet; ethanol), disinfect interior surfaces of cabinet; periodically remove the work surface and disinfect the periodically remove the work surface and disinfect the area beneath it (including the catch pan) and wipe the area beneath it (including the catch pan) and wipe the surface of the UV light with disinfectant.surface of the UV light with disinfectant.

Turn off the fluorescent light and cabinet blower when Turn off the fluorescent light and cabinet blower when appropriate appropriate

Turn on the UV light if appropriate (do not turn on Turn on the UV light if appropriate (do not turn on when people are working close by); UV must be tested when people are working close by); UV must be tested to ensure that it is emitting a germicidal wavelength.to ensure that it is emitting a germicidal wavelength.

Safe Work Practices for BSC Use

Do not store equipment or supplies inside the cabinetDo not store equipment or supplies inside the cabinet.. Do not use the top of the cabinet for storage. The HEPA Do not use the top of the cabinet for storage. The HEPA

filter could be damaged and the airflow disrupted.filter could be damaged and the airflow disrupted. Make sure the cabinet is level. If the cabinet base is Make sure the cabinet is level. If the cabinet base is

uneven, airflow can be affected.uneven, airflow can be affected. Never disengage the alarm. It indicates improper airflow Never disengage the alarm. It indicates improper airflow

and reduced performance which may endanger the and reduced performance which may endanger the researcher or the experiment.researcher or the experiment.

Never completely close the window sash with the motor Never completely close the window sash with the motor running as this condition may cause motor burnout.running as this condition may cause motor burnout.

Cabinets should be placed away from doors, windows, Cabinets should be placed away from doors, windows, vents or high traffic areas to reduce air turbulence.vents or high traffic areas to reduce air turbulence.

Do not store equipment or supplies inside the cabinetDo not store equipment or supplies inside the cabinet.. Do not use the top of the cabinet for storage. The HEPA Do not use the top of the cabinet for storage. The HEPA

filter could be damaged and the airflow disrupted.filter could be damaged and the airflow disrupted. Make sure the cabinet is level. If the cabinet base is Make sure the cabinet is level. If the cabinet base is

uneven, airflow can be affected.uneven, airflow can be affected. Never disengage the alarm. It indicates improper airflow Never disengage the alarm. It indicates improper airflow

and reduced performance which may endanger the and reduced performance which may endanger the researcher or the experiment.researcher or the experiment.

Never completely close the window sash with the motor Never completely close the window sash with the motor running as this condition may cause motor burnout.running as this condition may cause motor burnout.

Cabinets should be placed away from doors, windows, Cabinets should be placed away from doors, windows, vents or high traffic areas to reduce air turbulence.vents or high traffic areas to reduce air turbulence.


For BSC without fixed exhaust, the cabinet exhaust For BSC without fixed exhaust, the cabinet exhaust should have a twelve inch clearance from the ceiling for should have a twelve inch clearance from the ceiling for proper exhaust air flow. Also, allow a twelve inch proper exhaust air flow. Also, allow a twelve inch clearance on both sides of the cabinet for maintenance clearance on both sides of the cabinet for maintenance purposes.purposes.

Never operate a cabinet while a warning light or alarm Never operate a cabinet while a warning light or alarm is on.is on.

The operator should be seated with shoulders level with The operator should be seated with shoulders level with the bottom of the sash.the bottom of the sash.

Perform all work using a limited number of slow Perform all work using a limited number of slow movements. movements.

Keep all materials at least four inches inside the sash Keep all materials at least four inches inside the sash opening.opening.

To avoid excessive movements in and out of the cabinet, To avoid excessive movements in and out of the cabinet, discard pipettes into a tray, container or biohazard bag discard pipettes into a tray, container or biohazard bag within the cabinet .within the cabinet .

For BSC without fixed exhaust, the cabinet exhaust For BSC without fixed exhaust, the cabinet exhaust should have a twelve inch clearance from the ceiling for should have a twelve inch clearance from the ceiling for proper exhaust air flow. Also, allow a twelve inch proper exhaust air flow. Also, allow a twelve inch clearance on both sides of the cabinet for maintenance clearance on both sides of the cabinet for maintenance purposes.purposes.

Never operate a cabinet while a warning light or alarm Never operate a cabinet while a warning light or alarm is on.is on.

The operator should be seated with shoulders level with The operator should be seated with shoulders level with the bottom of the sash.the bottom of the sash.

Perform all work using a limited number of slow Perform all work using a limited number of slow movements. movements.

Keep all materials at least four inches inside the sash Keep all materials at least four inches inside the sash opening.opening.

To avoid excessive movements in and out of the cabinet, To avoid excessive movements in and out of the cabinet, discard pipettes into a tray, container or biohazard bag discard pipettes into a tray, container or biohazard bag within the cabinet .within the cabinet .


If a bunsen burner must be used, place it at the If a bunsen burner must be used, place it at the rear of the work area where the air turbulence rear of the work area where the air turbulence from the flame will have the least possible effect from the flame will have the least possible effect on the air stream. on the air stream.

All equipment which has come in contact with the All equipment which has come in contact with the biological agent should be decontaminated.biological agent should be decontaminated.

The cabinet should be allowed to run for at least The cabinet should be allowed to run for at least three minutes with no activity so that the three minutes with no activity so that the airborne contaminants will be purged from the airborne contaminants will be purged from the work area before removing equipment. work area before removing equipment.

After all items have been removed, wipe the After all items have been removed, wipe the interior surfaces with disinfectant.interior surfaces with disinfectant.

If a bunsen burner must be used, place it at the If a bunsen burner must be used, place it at the rear of the work area where the air turbulence rear of the work area where the air turbulence from the flame will have the least possible effect from the flame will have the least possible effect on the air stream. on the air stream.

All equipment which has come in contact with the All equipment which has come in contact with the biological agent should be decontaminated.biological agent should be decontaminated.

The cabinet should be allowed to run for at least The cabinet should be allowed to run for at least three minutes with no activity so that the three minutes with no activity so that the airborne contaminants will be purged from the airborne contaminants will be purged from the work area before removing equipment. work area before removing equipment.

After all items have been removed, wipe the After all items have been removed, wipe the interior surfaces with disinfectant.interior surfaces with disinfectant.

Biohazard Spill Control Inside a BSC

Keep the BSC on.Keep the BSC on. Put on protective gloves.Put on protective gloves. Spray & wipe walls, work surfaces, and Spray & wipe walls, work surfaces, and

equipment with decontamination equipment with decontamination solution.solution.

Flood tray top, drain pans, and catch Flood tray top, drain pans, and catch basins with decontamination solution.basins with decontamination solution.

Allow to stand for 20 minutes.Allow to stand for 20 minutes. Drain excess solution into cabinet base.Drain excess solution into cabinet base. Lift out tray and any removable exhaust Lift out tray and any removable exhaust

grille grille work.work.

Keep the BSC on.Keep the BSC on. Put on protective gloves.Put on protective gloves. Spray & wipe walls, work surfaces, and Spray & wipe walls, work surfaces, and

equipment with decontamination equipment with decontamination solution.solution.

Flood tray top, drain pans, and catch Flood tray top, drain pans, and catch basins with decontamination solution.basins with decontamination solution.

Allow to stand for 20 minutes.Allow to stand for 20 minutes. Drain excess solution into cabinet base.Drain excess solution into cabinet base. Lift out tray and any removable exhaust Lift out tray and any removable exhaust

grille grille work.work.

Biohazard Spill Control Inside a BSC

Clean top and bottom surfaces Clean top and bottom surfaces with sponge/cloth soaked in with sponge/cloth soaked in decontamination solution.decontamination solution.

Replace tray and grille work. Replace tray and grille work. Place everything that is Place everything that is

contaminated into autoclave contaminated into autoclave pan.pan.

Drain decontamination solution Drain decontamination solution from cabinet base into from cabinet base into AUTOCLAVABLE containers.AUTOCLAVABLE containers.

Clean top and bottom surfaces Clean top and bottom surfaces with sponge/cloth soaked in with sponge/cloth soaked in decontamination solution.decontamination solution.

Replace tray and grille work. Replace tray and grille work. Place everything that is Place everything that is

contaminated into autoclave contaminated into autoclave pan.pan.

Drain decontamination solution Drain decontamination solution from cabinet base into from cabinet base into AUTOCLAVABLE containers.AUTOCLAVABLE containers.

Biological Safety CabinetCertification

The cabinet must be certified when first installed The cabinet must be certified when first installed and then annually. It must be recertified and then annually. It must be recertified anytime it is moved even within the same room. anytime it is moved even within the same room. Before certification personnel arrive, remove all Before certification personnel arrive, remove all items from the cabinet and wipe it down with a items from the cabinet and wipe it down with a disinfectant. This will expedite the certification. disinfectant. This will expedite the certification. Any decontaminations, certifications, repairs or Any decontaminations, certifications, repairs or adjustments are to be made by qualified adjustments are to be made by qualified personnel.personnel.

The cabinet must be certified when first installed The cabinet must be certified when first installed and then annually. It must be recertified and then annually. It must be recertified anytime it is moved even within the same room. anytime it is moved even within the same room. Before certification personnel arrive, remove all Before certification personnel arrive, remove all items from the cabinet and wipe it down with a items from the cabinet and wipe it down with a disinfectant. This will expedite the certification. disinfectant. This will expedite the certification. Any decontaminations, certifications, repairs or Any decontaminations, certifications, repairs or adjustments are to be made by qualified adjustments are to be made by qualified personnel.personnel.

And remember……..

A laminar flow biological safety A laminar flow biological safety cabinet is a valuable supplement to cabinet is a valuable supplement to good sterile technique, good sterile technique, not a not a replacement for itreplacement for it. .

If the cabinet is not well understood If the cabinet is not well understood and operated correctly, it will not and operated correctly, it will not provide adequate protection for you provide adequate protection for you or the environment.or the environment.

A laminar flow biological safety A laminar flow biological safety cabinet is a valuable supplement to cabinet is a valuable supplement to good sterile technique, good sterile technique, not a not a replacement for itreplacement for it. .

If the cabinet is not well understood If the cabinet is not well understood and operated correctly, it will not and operated correctly, it will not provide adequate protection for you provide adequate protection for you or the environment.or the environment.

biohazrd a biological agent, such as an infectious microorganism, that constitutes a threat to...

Documents

low community risk risk

high community risk

environment risk group

risk of spread

classification risk

community risk slide

infectious agents

low individual