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

3. CLASSIFICATION Risk Group 1 Risk Group 1 Risk Group 2 Risk Group 2 Risk Group 3 Risk Group 4 Risk Group 1 Risk Group 1 Risk Group 2 Risk Group 2 Risk Group 3 Risk Group 4 No or low individual and community risk Moderate individual risk, low community risks High individual risk, low community risk High individual and community risk No or low individual and community risk Moderate individual risk, low community risks High individual risk, low community risk High individual and community risk

4. Risk Group 1 No or low individual and community risk A microorganism that is unlikely to cause disease in healthy human or animals

5. Risk Group 2 Moderate individual risk, low community risk Risk Group 2 infectious agents are pathogens that can cause human or animal disease but, under normal circumstances, are unlikely to be a serious hazard to laboratory workers, the community, or the environment Risk Group 2 infectious agents are pathogens that can cause human or animal disease but, under normal circumstances, are unlikely to be a serious hazard to laboratory workers, the community, or the environment Effective treatment and preventive measures are available and the risk of spread is limited. Effective treatment and preventive measures are available and the risk of spread is limited. Risk Group 2 infectious agents are pathogens that can cause human or animal disease but, under normal circumstances, are unlikely to be a serious hazard to laboratory workers, the community, or the environment Risk Group 2 infectious agents are pathogens that can cause human or animal disease but, under normal circumstances, are unlikely to be a serious hazard to laboratory workers, the community, or the environment Effective treatment and preventive measures are available and the risk of spread is limited. Effective treatment and preventive measures are available and the risk of spread is limited.

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

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

8. Risk Group 3(contd.) Risk Group 3 pathogens include Risk Group 3 pathogens include BACTERIAS such as Bacilluus, Brucella, Chlamydia, M. tuberculosis, Rickettsia, Salmonella typhi, Shigella dysentriae type 1, Yersinia pestis etc. BACTERIAS such as Bacilluus, Brucella, Chlamydia, M. tuberculosis, Rickettsia, Salmonella typhi, Shigella dysentriae type 1, Yersinia pestis etc. FUNGI such as Histoplasma, Blastomyces, etc. FUNGI such as Histoplasma, Blastomyces, etc. VIRUSES such as Hepatitis A,B,C,D,E virus, Herpes virus, HIV, HTLV, Rabies virus etc. VIRUSES such as Hepatitis A,B,C,D,E virus, Herpes virus, HIV, HTLV, Rabies virus etc.

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

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

11. BIOLOGICAL CONTAINMENT Any methods to contain organisms by creating biochemical barriers to prevent them from growing outside the laboratory. Or Any methods to contain organisms by creating biochemical barriers to prevent them from growing outside the laboratory. Or system designed to prevent the accidental release of biological material. system designed to prevent the accidental release of biological material. Any methods to contain organisms by creating biochemical barriers to prevent them from growing outside the laboratory. Or Any methods to contain organisms by creating biochemical barriers to prevent them from growing outside the laboratory. Or system designed to prevent the accidental release of biological material. system designed to prevent the accidental release of biological material.

12. Biosafety Level 1 (Containment Level 1) No special facilities, equipment or procedures are required No special facilities, equipment or procedures 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 procedures are required No special facilities, equipment or procedures 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.

13. 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.

14. 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.

15. 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.

16. 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.

17. Biosafety level 1 Lab. 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

18. Biosafety level 1 Lab.

19. 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.

20. BIOHAZARD ADMITTANCE TO AUTHORIZED PERSONNEL ONLY Biosafety Level: _________________________________ Responsible Investigator: _________________________ In case of emergency call: ________________________ Daytime phone: __________Home phone: ___________ Authorization for entrance must be obtained from the Responsible Investigator named above BIOHAZARD ADMITTANCE TO AUTHORIZED PERSONNEL ONLY Biosafety Level: _________________________________ Responsible Investigator: _________________________ In case of emergency call: ________________________ Daytime phone: __________Home phone: ___________ Authorization for entrance must be obtained from the Responsible Investigator named above

21. 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

22. 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 biological safety cabinet for organism manipulations. Certified HEPA filtered class I or II biological safety cabinet for organism manipulations. All contaminated material to be properly decontaminated All contaminated material to be 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 biological safety cabinet for organism manipulations. Certified HEPA filtered class I or II biological safety cabinet for organism manipulations. All contaminated material to be properly decontaminated All contaminated material to be properly decontaminated

23. Biosafety level 2 A preplacement health check is necessary. The persons 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 persons 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.

24. Biosafety level 2 Lab.

25. 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.

26. 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.

27. 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.

28. Biosafety level 3 Air exhausted through a dedicated exhaust or HEPA filtration system Air exhausted through a dedicated exhaust or HEPA filtration system certified HEPA filtered class II biological safety cabinet for organism manipulations certified HEPA filtered class II biological safety cabinet for organism manipulations Air exhausted through a dedicated exhaust or HEPA filtration system Air exhausted through a dedicated exhaust or HEPA filtration system certified HEPA filtered class II biological safety cabinet for organism manipulations certified HEPA filtered class II biological safety cabinet for organism manipulations

29. Biosafety level 3 Lab.

30. 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.

31. 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

32. BIOSAFETY LEVEL1234 Isolation of laboratoryNONOYESYES Room sealable for decontaminationNONOYESYES Ventilation inward airflow NODEYESYES controlled ventilating system NODEYESYES HEPA-filtered air exhaust NONO YES/ NO YES Double-door entry NONOYESYES AirlockNONONOYES BIOSAFETY REQUIREMENTS

33. Airlock with shower NONONOYES AnteroomNONOYES--- Anteroom with shower NONO NO/ YES YES Effluent treatment NONO NO/ YES YES Autoclave on site NODEYESYES in laboratory room NONODEYES double-endedNONODEYES Biological safety cabinets NODEYESYES Personnel safety monitoring capability NONODEYES

34. Biological Safety Cabinets (BSC) Containment and protection devices used in laboratories working with biological agents with a primary purpose of protecting the laboratory worker and the environment from viable organisms.

35. Class I Biological Safety Cabinets Class I BSC is a ventilated cabinet in which r. Class I BSC is a ventilated cabinet 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 cabinet in which r. Class I BSC is a ventilated cabinet 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

36. Class I Biological Safety Cabinet

37. Class I Biological Safety Cabinets

38. Class II Biological Safety Cabinets Class II BSCs are Laminar Flow Biological Safety Cabinets that protect personnel, product, and environment. Class II BSCs are Laminar Flow Biological Safety Cabinets that protect personnel, product, and environment. They provide an inward airflow to protect personnel, a downward flow of HEPA filtered air to the work area to protect the product, and then exhaust HEPA filtered air to protect the environment from particulate and aerosol hazards. They provide an inward airflow to protect personnel, a downward flow of HEPA filtered air to the work area to protect the product, and then exhaust HEPA filtered air to protect the environment from particulate and aerosol 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 BSCs are Laminar Flow Biological Safety Cabinets that protect personnel, product, and environment. Class II BSCs are Laminar Flow Biological Safety Cabinets that protect personnel, product, and environment. They provide an inward airflow to protect personnel, a downward flow of HEPA filtered air to the work area to protect the product, and then exhaust HEPA filtered air to protect the environment from particulate and aerosol hazards. They provide an inward airflow to protect personnel, a downward flow of HEPA filtered air to the work area to protect the product, and then exhaust HEPA filtered air to protect the environment from particulate and aerosol 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

39. Class II Type A 1 BSC Class || Type A1 BSC offer personnel protection through negative pressure airflow into the cabinet. Class || Type A1 BSC offer personnel protection through negative pressure airflow into the cabinet. To protect the product, the work area in the cabinet is continuously bathed with ultra- clean air provided by the supply HEPA filter. To protect the product, the work area in the cabinet is continuously bathed with ultra- clean air provided by the supply HEPA filter. Maintain a minimum average face velocity of 0.38 m/s Maintain a minimum average face velocity of 0.38 m/s Approximately 70% of the air of each cycle is recirculated through this supply HEPA filter. Approximately 70% of the air of each cycle is recirculated through this supply HEPA filter. The remaining air is discharged from the hood through the exhaust HEPA filter, protecting the environment. The remaining air is discharged from the hood through the exhaust HEPA filter, protecting the environment. Exhaust to room or thimble connection vented to the outside Class || Type A1 BSC offer personnel protection through negative pressure airflow into the cabinet. Class || Type A1 BSC offer personnel protection through negative pressure airflow into the cabinet. To protect the product, the work area in the cabinet is continuously bathed with ultra- clean air provided by the supply HEPA filter. To protect the product, the work area in the cabinet is continuously bathed with ultra- clean air provided by the supply HEPA filter. Maintain a minimum average face velocity of 0.38 m/s Maintain a minimum average face velocity of 0.38 m/s Approximately 70% of the air of each cycle is recirculated through this supply HEPA filter. Approximately 70% of the air of each cycle is recirculated through this supply HEPA filter. The remaining air is discharged from the hood through the exhaust HEPA filter, protecting the environment. The remaining air is discharged from the hood through the exhaust HEPA filter, protecting the environment. Exhaust to room or thimble connection vented to the outside

40. Class II Type A 1 BSC A- Front Opening B- Sash C- Exhaust HEPA Filter D- Rear Plenum E- Supply HEPA Filter F- Blower

41. Class II Type A 1 BSC

42. Class 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

43. Class II Type A 2 BSC A. narrow front opening B. sash C. exhaust HEPA filter D. supply HEPA filter E. negative pressure exhaust plenum F. supply blower A. narrow front opening B. sash C. exhaust HEPA filter D. supply HEPA filter E. negative pressure exhaust plenum F. supply blower

44. Class II Type A 2 BSC

45. Class 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 cell culture or microbial systems require both biological and chemical 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 cell culture or microbial systems require both biological and chemical containment.

46. Class II Type B 1 BSC The cabinet supply blowers draw room air through the front grille and a portion of the cabinets 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 cabinets 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.

47. Class II Type B 1 BSC 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 toxic chemicals and trace amounts of radionuclides Suitable for work with low levels of volatile toxic chemicals and trace amounts of 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 toxic chemicals and trace amounts of radionuclides Suitable for work with low levels of volatile toxic chemicals and trace amounts of radionuclides

48. Class II Type B 1 BSC A- Front Opening B- Sash C- Exhaust HEPA Filter D- Supply Plenum E- Supply HEPA Filter F- Blower

49. Class II Type B 1 BSC

50. Class II Type B 2 BSC This is a total-exhaust cabinet; no air is recirculated within it This is a total-exhaust cabinet; no air is recirculated within it This cabinet provides simultaneous primary biological and chemical containment. This cabinet provides simultaneous primary biological and chemical containment. The supply blower draws in room air or outside air at the top of the cabinet, passes it through a HEPA filter and down into the work area of the cabinet. The supply blower draws in room air or outside air at the top of the cabinet, passes it through a HEPA filter and down into the work 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 recirculated within it This is a total-exhaust cabinet; no air is recirculated within it This cabinet provides simultaneous primary biological and chemical containment. This cabinet provides simultaneous primary biological and chemical containment. The supply blower draws in room air or outside air at the top of the cabinet, passes it through a HEPA filter and down into the work area of the cabinet. The supply blower draws in room air or outside air at the top of the cabinet, passes it through a HEPA filter and down into the work area of the cabinet. Maintain a minimum average inflow velocity of at least 0.51m/s at the face opening of the cabinet

51. 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.

52. Class II Type B 2 BSC Schematic. A. front opening B. sash C. exhaust HEPA filter D. supply HEPA filter E. negative pressure exhaust plenum F. supply blower G. filter screen The cabinet exhaust needs to be connected to the building exhaust system.

53. Class II Type B 2 BSC

54. Class II BSC Modifications The front sash can be modified by the manufacturer to accommodate a microscope A rigid plate with arm holes can be added if needed. The work surface can be designed to accept a centrifuge, or other equipment that requires containment Good cabinet design, microbiological aerosol tracer testing of the modification, and appropriate certification are required to ensure that the basic BSC systems operate properly after modification. Maximum containment potential is achieved only through strict adherence to proper practices and procedures. The front sash can be modified by the manufacturer to accommodate a microscope A rigid plate with arm holes can be added if needed. The work surface can be designed to accept a centrifuge, or other equipment that requires containment Good cabinet design, microbiological aerosol tracer testing of the modification, and appropriate certification are required to ensure that the basic BSC systems operate properly after modification. Maximum containment potential is achieved only through strict adherence to proper practices and procedures. Class II BSCs can be modified to accommodate special tasks:

55. Class III BSC or Glovebox The Class III BSC was designed for work with Biosafety Level 4 microbiological agents and provides maximum protection to the environment and the worker. 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 that is accessible through the cabinet floor or double door pass-through box such as an autoclave that can be decontaminated between uses. Reversing that process allows for safe removal of materials from the Class III BSC. 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 discharge to the outdoors. Airflow is maintained by a dedicated independent exhaust system exterior to the cabinet which keeps the cabinet under negative pressure The Class III BSC was designed for work with Biosafety Level 4 microbiological agents and provides maximum protection to the environment and the worker. 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 that is accessible through the cabinet floor or double door pass-through box such as an autoclave that can be decontaminated between uses. Reversing that process allows for safe removal of materials from the Class III BSC. 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 discharge to the outdoors. Airflow is maintained by a dedicated independent exhaust system exterior to the cabinet which keeps the cabinet under negative pressure

56. 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

57. 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.

58. Class III BSC or Glove boxes

59. Class III BSC Schematic A- Glove Ports B- Sash C- Exhaust HEPA Filter D- Supply HEPA Filter E- Double Ended Autoclave or Pass Through 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.

60. BSCFACE VELOCITY (m/s) AIRFLOW (%) RECIRCUL- ATED AIRFLOW (%) EXHAUST- ED EXHAUST SYSTEM Class I0.360100Hard duct Class || A1 0.38–0.517030Exhaust to room or thimble connection Class || A2 vented to the outside 0.517030 Exhaust to room or thimble connection Class || B1 0.513070Hard duct Class || B2 0.510100Hard duct Class |||NA0100Hard duct

61. Start-up procedures Turn off UV lights if in use and ensure that the sash is in the appropriate position. Turn off UV lights if in use and ensure that the sash is 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 obstructions. Check the air intake and exhaust grilles for obstructions. If the cabinet is equipped with an alarm, test the alarm and switch it to the "on" position. If the cabinet is equipped with an alarm, test the alarm and switch it to the "on" position. Confirm inward airflow by holding a tissue at the middle of the edge of the viewing panel and ensuring that it is drawn in. Confirm inward airflow by holding a tissue at the middle of the edge of the viewing panel and ensuring that it is drawn in. Disinfect the interior surfaces with a suitable, noncorrosive disinfectant. Disinfect the interior surfaces with a suitable, noncorrosive disinfectant. Assemble all materials required for the procedure and load them into the cabinet; do not obstruct the air grilles; Assemble all materials required for the procedure and load them into the cabinet; do not obstruct the air grilles; Wait 5 minutes to purge airborne contaminants from the work area. Wait 5 minutes to purge airborne contaminants from the work area. Turn off UV lights if in use and ensure that the sash is in the appropriate position. Turn off UV lights if in use and ensure that the sash is 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 obstructions. Check the air intake and exhaust grilles for obstructions. If the cabinet is equipped with an alarm, test the alarm and switch it to the "on" position. If the cabinet is equipped with an alarm, test the alarm and switch it to the "on" position. Confirm inward airflow by holding a tissue at the middle of the edge of the viewing panel and ensuring that it is drawn in. Confirm inward airflow by holding a tissue at the middle of the edge of the viewing panel and ensuring that it is drawn in. Disinfect the interior surfaces with a suitable, noncorrosive disinfectant. Disinfect the interior surfaces with a suitable, noncorrosive disinfectant. Assemble all materials required for the procedure and load them into the cabinet; do not obstruct the air grilles; Assemble all materials required for the procedure and load them into the cabinet; do not obstruct the air grilles; Wait 5 minutes to purge airborne contaminants from the work area. Wait 5 minutes to purge airborne contaminants from the work area.

62. 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 from the cabinet. Close or cover open containers before removing them from the cabinet. Surface disinfect objects in contact with contaminated material before removal from the cabinet. Surface disinfect objects in contact with contaminated material before removal from the cabinet. Remove contaminated gloves and dispose of them as appropriate; wash hands. Remove contaminated gloves and dispose of them as appropriate; wash hands. Using a suitable non-corrosive disinfectant (e.g., 70% ethanol), disinfect interior surfaces of cabinet; periodically remove the work surface and disinfect the area beneath it (including the catch pan) and wipe the surface of the UV light with disinfectant. Using a suitable non-corrosive disinfectant (e.g., 70% ethanol), disinfect interior surfaces of cabinet; periodically remove the work surface and disinfect the area beneath it (including the catch pan) and wipe the surface of the UV light with disinfectant. Turn off the fluorescent light and cabinet blower when appropriate Turn off the fluorescent light and cabinet blower when appropriate Turn on the UV light if appropriate (do not turn on when people are working close by); UV must be tested to ensure that it is emitting a germicidal wavelength. Turn on the UV light if appropriate (do not turn on when people are working close by); UV must be tested 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 from the cabinet. Close or cover open containers before removing them from the cabinet. Surface disinfect objects in contact with contaminated material before removal from the cabinet. Surface disinfect objects in contact with contaminated material before removal from the cabinet. Remove contaminated gloves and dispose of them as appropriate; wash hands. Remove contaminated gloves and dispose of them as appropriate; wash hands. Using a suitable non-corrosive disinfectant (e.g., 70% ethanol), disinfect interior surfaces of cabinet; periodically remove the work surface and disinfect the area beneath it (including the catch pan) and wipe the surface of the UV light with disinfectant. Using a suitable non-corrosive disinfectant (e.g., 70% ethanol), disinfect interior surfaces of cabinet; periodically remove the work surface and disinfect the area beneath it (including the catch pan) and wipe the surface of the UV light with disinfectant. Turn off the fluorescent light and cabinet blower when appropriate Turn off the fluorescent light and cabinet blower when appropriate Turn on the UV light if appropriate (do not turn on when people are working close by); UV must be tested to ensure that it is emitting a germicidal wavelength. Turn on the UV light if appropriate (do not turn on when people are working close by); UV must be tested to ensure that it is emitting a germicidal wavelength.

63. Safe Work Practices for BSC Use Do not store equipment or supplies inside the cabinet. Do not store equipment or supplies inside the cabinet. Do not use the top of the cabinet for storage. The HEPA filter could be damaged and the airflow disrupted. Do not use the top of the cabinet for storage. The HEPA filter could be damaged and the airflow disrupted. Make sure the cabinet is level. If the cabinet base is uneven, airflow can be affected. Make sure the cabinet is level. If the cabinet base is uneven, airflow can be affected. Never disengage the alarm. It indicates improper airflow and reduced performance which may endanger the researcher or the experiment. Never disengage the alarm. It indicates improper airflow and reduced performance which may endanger the researcher or the experiment. Never completely close the window sash with the motor running as this condition may cause motor burnout. Never completely close the window sash with the motor running as this condition may cause motor burnout. Cabinets should be placed away from doors, windows, vents or high traffic areas to reduce air turbulence. Cabinets should be placed away from doors, windows, vents or high traffic areas to reduce air turbulence. Do not store equipment or supplies inside the cabinet. Do not store equipment or supplies inside the cabinet. Do not use the top of the cabinet for storage. The HEPA filter could be damaged and the airflow disrupted. Do not use the top of the cabinet for storage. The HEPA filter could be damaged and the airflow disrupted. Make sure the cabinet is level. If the cabinet base is uneven, airflow can be affected. Make sure the cabinet is level. If the cabinet base is uneven, airflow can be affected. Never disengage the alarm. It indicates improper airflow and reduced performance which may endanger the researcher or the experiment. Never disengage the alarm. It indicates improper airflow and reduced performance which may endanger the researcher or the experiment. Never completely close the window sash with the motor running as this condition may cause motor burnout. Never completely close the window sash with the motor running as this condition may cause motor burnout. Cabinets should be placed away from doors, windows, vents or high traffic areas to reduce air turbulence. Cabinets should be placed away from doors, windows, vents or high traffic areas to reduce air turbulence.

64. Safe Work Practices for BSC Use For BSC without fixed exhaust, the cabinet exhaust should have a twelve inch clearance from the ceiling for proper exhaust air flow. Also, allow a twelve inch clearance on both sides of the cabinet for maintenance purposes. For BSC without fixed exhaust, the cabinet exhaust should have a twelve inch clearance from the ceiling for proper exhaust air flow. Also, allow a twelve inch clearance on both sides of the cabinet for maintenance purposes. Never operate a cabinet while a warning light or alarm is on. Never operate a cabinet while a warning light or alarm is on. The operator should be seated with shoulders level with the bottom of the sash. The operator should be seated with shoulders level with the bottom of the sash. Perform all work using a limited number of slow movements. Perform all work using a limited number of slow movements. Keep all materials at least four inches inside the sash opening. Keep all materials at least four inches inside the sash opening. To avoid excessive movements in and out of the cabinet, discard pipettes into a tray, container or biohazard bag within the cabinet. To avoid excessive movements in and out of the cabinet, discard pipettes into a tray, container or biohazard bag within the cabinet. For BSC without fixed exhaust, the cabinet exhaust should have a twelve inch clearance from the ceiling for proper exhaust air flow. Also, allow a twelve inch clearance on both sides of the cabinet for maintenance purposes. For BSC without fixed exhaust, the cabinet exhaust should have a twelve inch clearance from the ceiling for proper exhaust air flow. Also, allow a twelve inch clearance on both sides of the cabinet for maintenance purposes. Never operate a cabinet while a warning light or alarm is on. Never operate a cabinet while a warning light or alarm is on. The operator should be seated with shoulders level with the bottom of the sash. The operator should be seated with shoulders level with the bottom of the sash. Perform all work using a limited number of slow movements. Perform all work using a limited number of slow movements. Keep all materials at least four inches inside the sash opening. Keep all materials at least four inches inside the sash opening. To avoid excessive movements in and out of the cabinet, discard pipettes into a tray, container or biohazard bag within the cabinet. To avoid excessive movements in and out of the cabinet, discard pipettes into a tray, container or biohazard bag within the cabinet.

65. Safe Work Practices for BSC Use If a bunsen burner must be used, place it at the rear of the work area where the air turbulence from the flame will have the least possible effect on the air stream. If a bunsen burner must be used, place it at the rear of the work area where the air turbulence from the flame will have the least possible effect on the air stream. All equipment which has come in contact with the biological agent should be decontaminated. All equipment which has come in contact with the biological agent should be decontaminated. The cabinet should be allowed to run for at least three minutes with no activity so that the airborne contaminants will be purged from the work area before removing equipment. The cabinet should be allowed to run for at least three minutes with no activity so that the airborne contaminants will be purged from the work area before removing equipment. After all items have been removed, wipe the interior surfaces with disinfectant. After all items have been removed, wipe the interior surfaces with disinfectant. If a bunsen burner must be used, place it at the rear of the work area where the air turbulence from the flame will have the least possible effect on the air stream. If a bunsen burner must be used, place it at the rear of the work area where the air turbulence from the flame will have the least possible effect on the air stream. All equipment which has come in contact with the biological agent should be decontaminated. All equipment which has come in contact with the biological agent should be decontaminated. The cabinet should be allowed to run for at least three minutes with no activity so that the airborne contaminants will be purged from the work area before removing equipment. The cabinet should be allowed to run for at least three minutes with no activity so that the airborne contaminants will be purged from the work area before removing equipment. After all items have been removed, wipe the interior surfaces with disinfectant. After all items have been removed, wipe the interior surfaces with disinfectant.

66. 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 equipment with decontamination solution. Spray & wipe walls, work surfaces, and equipment with decontamination solution. Flood tray top, drain pans, and catch basins with decontamination solution. Flood tray top, drain pans, and catch 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 grille work. Lift out tray and any removable exhaust grille work. Keep the BSC on. Keep the BSC on. Put on protective gloves. Put on protective gloves. Spray & wipe walls, work surfaces, and equipment with decontamination solution. Spray & wipe walls, work surfaces, and equipment with decontamination solution. Flood tray top, drain pans, and catch basins with decontamination solution. Flood tray top, drain pans, and catch 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 grille work. Lift out tray and any removable exhaust grille work.

67. Biohazard Spill Control Inside a BSC Clean top and bottom surfaces with sponge/cloth soaked in decontamination solution. Clean top and bottom surfaces with sponge/cloth soaked in decontamination solution. Replace tray and grille work. Replace tray and grille work. Place everything that is contaminated into autoclave pan. Place everything that is contaminated into autoclave pan. Drain decontamination solution from cabinet base into AUTOCLAVABLE containers. Drain decontamination solution from cabinet base into AUTOCLAVABLE containers. Clean top and bottom surfaces with sponge/cloth soaked in decontamination solution. Clean top and bottom surfaces with sponge/cloth soaked in decontamination solution. Replace tray and grille work. Replace tray and grille work. Place everything that is contaminated into autoclave pan. Place everything that is contaminated into autoclave pan. Drain decontamination solution from cabinet base into AUTOCLAVABLE containers. Drain decontamination solution from cabinet base into AUTOCLAVABLE containers.

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

69. And remember…….. A laminar flow biological safety cabinet is a valuable supplement to good sterile technique, not a replacement for it. A laminar flow biological safety cabinet is a valuable supplement to good sterile technique, not a replacement for it. If the cabinet is not well understood and operated correctly, it will not provide adequate protection for you or the environment. If the cabinet is not well understood and operated correctly, it will not provide adequate protection for you or the environment. A laminar flow biological safety cabinet is a valuable supplement to good sterile technique, not a replacement for it. A laminar flow biological safety cabinet is a valuable supplement to good sterile technique, not a replacement for it. If the cabinet is not well understood and operated correctly, it will not provide adequate protection for you or the environment. If the cabinet is not well understood and operated correctly, it will not provide adequate protection for you or the environment.