1. Containment Equipment and NSF 49

2. Containment – BMBL Primary containment is the protection of personnel and the immediate lab environment from exposure to infectious agents, and is provided by good microbiological technique and the use of appropriate safety equipment. Safety equipment (primary barriers) includes biological safety cabinets, enclosed containers, and other engineering controls (i.e. safety centrifuge cups) designed to remove or minimize exposure hazards to biological materials. Primary containment is the protection of personnel and the immediate lab environment from exposure to infectious agents, and is provided by good microbiological technique and the use of appropriate safety equipment. Safety equipment (primary barriers) includes biological safety cabinets, enclosed containers, and other engineering controls (i.e. safety centrifuge cups) designed to remove or minimize exposure hazards to biological materials.

3. Containment – BMBL Note: If a containment device such as a biological safety cabinet is impractical then personal protective equipment may form the primary barrier between personnel and the infectious materials. Ex. Certain animal studies, agent production activities, and activities related to the maintenance, service or support of the lab facility. Note: If a containment device such as a biological safety cabinet is impractical then personal protective equipment may form the primary barrier between personnel and the infectious materials. Ex. Certain animal studies, agent production activities, and activities related to the maintenance, service or support of the lab facility.

4. Biological Safety Cabinets (BSCs) Primary means of containment for infectious splashes or aerosols Can provide personnel, product, and environmental protection provided appropriate use and cabinet maintenance Not to be confused with horizontal laminar flow (clean benches) or vertical laminar flow hoods BSCs described in Appendix A of the BMBL

5. Biosafety Cabinets (BSCs)

6. BSCs – Recommendations and Guidance NSF/ANSI 49 – Class II (laminar flow) Biosafety Cabinetry (2002) CDC/NIH Primary Containment for Biohazards – Selection, Installation and Use of Biological Safety Cabinets (2000)CDC/NIH Primary Containment for Biohazards – Selection, Installation and Use of Biological Safety Cabinets (2000) BMBL 5 th Edition (2007), Appendix ABMBL 5 th Edition (2007), Appendix A Institutional Biological Safety Cabinet Guidance Documents NSF/ANSI 49 – Class II (laminar flow) Biosafety Cabinetry (2002) CDC/NIH Primary Containment for Biohazards – Selection, Installation and Use of Biological Safety Cabinets (2000)CDC/NIH Primary Containment for Biohazards – Selection, Installation and Use of Biological Safety Cabinets (2000) BMBL 5 th Edition (2007), Appendix ABMBL 5 th Edition (2007), Appendix A Institutional Biological Safety Cabinet Guidance Documents

7. BSCs – Class I

8. BSCs – Class II

9. BSCs – Class III

10. Has HEPA filtered downflow air that is a portion of the mixed downflow and inflow air May exhaust HEPA filtered air back into the laboratory or to the exterior of building through an exhaust canopy Has HEPA filtered downflow air that is a portion of the mixed downflow and inflow air May exhaust HEPA filtered air back into the laboratory or to the exterior of building through an exhaust canopy Class II A2

11. Biological Safety Cabinet Class II A2

13. Has HEPA filtered downflow air drawn from the laboratory or the outside air (i.e., no recirculation) Class II B2 cabinets are always hard ducted for exhaust to outside of building Has HEPA filtered downflow air drawn from the laboratory or the outside air (i.e., no recirculation) Class II B2 cabinets are always hard ducted for exhaust to outside of building Class II B2

14. Biological Safety Cabinet Class II B2

15. BSC must be turned on at least 5 minutes prior to starting work Personnel working within BSCs should wear lab coats, as well as appropriate gloves and safety glasses BSC work surfaces must be decontaminated with an appropriate disinfectant at the end of each work operation BSC must be turned on at least 5 minutes prior to starting work Personnel working within BSCs should wear lab coats, as well as appropriate gloves and safety glasses BSC work surfaces must be decontaminated with an appropriate disinfectant at the end of each work operation Safe Use of BSCs

16. Avoid excess motion and be aware of drafts in room that may interfere with normal air flow All cabinets must be certified for use on annual basis when first installed, moved, or repaired Avoid excess motion and be aware of drafts in room that may interfere with normal air flow All cabinets must be certified for use on annual basis when first installed, moved, or repaired Safe Use of BSCs

17. BSC Types and Operation YES3,4YESSupply air inlets and exhaust through double HEPA or HEPA/incineration combo N/A CLASS III YES2,3YESNo recirculation; total exhaust via HEPA & ducted 100CLASS II TYPE B2 YES2,3LOW LEVELS 30% recirculated through HEPA; exhaust via HEPA & ducted 100CLASS II TYPE B1 YES2,3LOW LEVELS Same as IIA1; but plena under neg. pressure; exhaust through HEPA or ducted 100CLASS II TYPE A2 YES2,3NO70% recirculated through HEPA; exhaust through HEPA 75CLASS II TYPE A1 NO2,3NOIn at front; rear & top through HEPA75 CLASS I PRODUCT PROTEC- TION BSL’sRAM or CHEMS ? AIRFLOW PATTERN FACE VELOCITY (LFPM) TYPE

18. HEPA Filters Removes particles > 0.3µm (includes all bacteria, spores, and viruses) to 99.97% efficiency - inertia, interception, diffusion, and electrostatic attachment The filter medium is borosilicate fibers treated with water-repellant binder Pleated to provide more surface area of filter, with separators to prevent collapse from air flow Removes particles > 0.3µm (includes all bacteria, spores, and viruses) to 99.97% efficiency - inertia, interception, diffusion, and electrostatic attachment The filter medium is borosilicate fibers treated with water-repellant binder Pleated to provide more surface area of filter, with separators to prevent collapse from air flow

19. BSCs – General Use Provide personnel, product and environmental protection for the use infectious biological agents in appropriate BSCs; can also be used to provide sterile environment. Leave unit on 24/7 or let run for 5 minutes prior to work. Do not block baffle openings or place bulky items in the cabinet that will prevent air from entering the baffle opening. Conduct work at least six inches from the edge of the hood at appropriate sash height. Keep cabinet clean and uncluttered. Wipe up spills immediately w/ 70% ETOH or other disinfectant. UV lights can be used but not generally recommended. Chemical disinfection is preferred. Be aware that drafts from open windows, open doors, fans, air conditioners, or high traffic walkways may interfere with normal cabinet air flow. Decontaminate all materials prior to exit from unit; including waste. No open flames in cabinet! Provide personnel, product and environmental protection for the use infectious biological agents in appropriate BSCs; can also be used to provide sterile environment. Leave unit on 24/7 or let run for 5 minutes prior to work. Do not block baffle openings or place bulky items in the cabinet that will prevent air from entering the baffle opening. Conduct work at least six inches from the edge of the hood at appropriate sash height. Keep cabinet clean and uncluttered. Wipe up spills immediately w/ 70% ETOH or other disinfectant. UV lights can be used but not generally recommended. Chemical disinfection is preferred. Be aware that drafts from open windows, open doors, fans, air conditioners, or high traffic walkways may interfere with normal cabinet air flow. Decontaminate all materials prior to exit from unit; including waste. No open flames in cabinet!

20. BSCs – Installation Considerations Ducting - If necessary depending on type (i.e. B1, B2) Plumbing - No combustible gas! Vacuum only. Electrical -Outlet above or below unit for access purposes during field certification process Location -BSCs should be located in lab where airflow patterns are not disrupted (i.e. away from doors, drafts, etc.) Ducting - If necessary depending on type (i.e. B1, B2) Plumbing - No combustible gas! Vacuum only. Electrical -Outlet above or below unit for access purposes during field certification process Location -BSCs should be located in lab where airflow patterns are not disrupted (i.e. away from doors, drafts, etc.)

21. BSCs – Design, Operation, and Certification NSF 49 – Basic requirements for design, construction, and performance of BSCs; standards for performance testing and field certification testing Installation recommendations Reliable operation Durability and structural stability Cleanability Limitations on noise and vibration Motor/blower performance Illumination NSF 49 – Basic requirements for design, construction, and performance of BSCs; standards for performance testing and field certification testing Installation recommendations Reliable operation Durability and structural stability Cleanability Limitations on noise and vibration Motor/blower performance Illumination

22. NSF 49, Annex F – Field Tests Certification performed at time of installation, removal of any panels, or after relocation; annually thereafter recommended REQUIRED: Downflow velocity profile test Inflow velocity test Airflow smoke patterns test HEPA filter leak test Cabinet integrity tests (Type A1 only) Site installation assessment tests OPTIONAL: Lighting intensity Vibration and noise level Electrical leakage, ground circuit resistance, polarity tests Certification performed at time of installation, removal of any panels, or after relocation; annually thereafter recommended REQUIRED: Downflow velocity profile test Inflow velocity test Airflow smoke patterns test HEPA filter leak test Cabinet integrity tests (Type A1 only) Site installation assessment tests OPTIONAL: Lighting intensity Vibration and noise level Electrical leakage, ground circuit resistance, polarity tests

23. Downflow Velocity Profile Test Purpose: measure velocity of air moving through workspace at 4 inches above bottom edge of window sash Measuring apparatus: thermal anemometer Acceptance: avg downflow velocity within +/- 5 ft/min of manufacturers specifications; within +/- 16 ft/min for any individual point Purpose: measure velocity of air moving through workspace at 4 inches above bottom edge of window sash Measuring apparatus: thermal anemometer Acceptance: avg downflow velocity within +/- 5 ft/min of manufacturers specifications; within +/- 16 ft/min for any individual point

24. Inflow Velocity Test Purpose: measure velocity of air moving through unit opening (face velocity) Measuring apparatus: thermal anemometer; direct inflow measurement instrument Acceptance: avg velocity within +/- 5 ft/min of normal flow for cabinet type Purpose: measure velocity of air moving through unit opening (face velocity) Measuring apparatus: thermal anemometer; direct inflow measurement instrument Acceptance: avg velocity within +/- 5 ft/min of normal flow for cabinet type

25. Air Smoke Patterns Test Purpose: measure airflow along the entire perimeter of the work access opening is inward, airflow within the work area is downward with no dead spots or refluxing, ambient air does not pass on or over the work surface, and there is no escape to the outside of the cabinet at the sides and top of the window Measuring apparatus: visible cold smoke such as titanium tetrachloride Acceptance: visual evidence verifying above purpose Purpose: measure airflow along the entire perimeter of the work access opening is inward, airflow within the work area is downward with no dead spots or refluxing, ambient air does not pass on or over the work surface, and there is no escape to the outside of the cabinet at the sides and top of the window Measuring apparatus: visible cold smoke such as titanium tetrachloride Acceptance: visual evidence verifying above purpose

26. HEPA Filter Leak Test Purpose: ensure integrity of downflow and exhaust HEPA’s, filter housing, and filter mounting frame Measuring apparatus: aerosol photometer capable of measuring 10 µg/L of polydisperse dioctylphthalate (DOP)particles; other aerosol generator w/ pressure gauge Acceptance: sustained aerosol penetration shall not exceed 0.01% of the upstream concentration at any point Purpose: ensure integrity of downflow and exhaust HEPA’s, filter housing, and filter mounting frame Measuring apparatus: aerosol photometer capable of measuring 10 µg/L of polydisperse dioctylphthalate (DOP)particles; other aerosol generator w/ pressure gauge Acceptance: sustained aerosol penetration shall not exceed 0.01% of the upstream concentration at any point

27. Cabinet Integrity Tests Pressure Decay/Soap bubble: performed to determine if exterior surfaces of all plenums, welds, gaskets, and plenum penetrations or seals are free of leaks Pressurize cabinet to 2” wg for 30 minutes with no soap bubbles or leakage of <0.2”wg Pressure Decay/Soap bubble: performed to determine if exterior surfaces of all plenums, welds, gaskets, and plenum penetrations or seals are free of leaks Pressurize cabinet to 2” wg for 30 minutes with no soap bubbles or leakage of <0.2”wg

28. Site Installation Assessment Tests Airflow alarm Sash alarm Interlocks Exhaust system performance / canopy connection Airflow alarm Sash alarm Interlocks Exhaust system performance / canopy connection

29. Understanding & Using Protective Hoods During Animal Husbandry Activities

30. Biosafety cabinets (BSC) Chemical fume hoods Animal cage changing stations Laminar flow hoods Biosafety cabinets (BSC) Chemical fume hoods Animal cage changing stations Laminar flow hoods Types of Hoods at UTHSC-H

31. Chemical Fume Hood

32. Protective barrier for working with hazardous chemicals Has no HEPA filtration- exhausted directly out of building Chemical fume hoods are not BSCs!!! Protects worker, but not work environment Protection from animal dander Ideal for those with allergies Protective barrier for working with hazardous chemicals Has no HEPA filtration- exhausted directly out of building Chemical fume hoods are not BSCs!!! Protects worker, but not work environment Protection from animal dander Ideal for those with allergies What is a Chemical Fume Hood?

33. Conduct work at least six inches from the edge of the hood Lower the sash to a reasonable height to protect yourself Keep hood clean and uncluttered Have a general awareness of the operation of hood and Be aware of any visual or audible differences that may indicate a change in function Conduct work at least six inches from the edge of the hood Lower the sash to a reasonable height to protect yourself Keep hood clean and uncluttered Have a general awareness of the operation of hood and Be aware of any visual or audible differences that may indicate a change in function Safe Use of Chem Fume Hood

34. Allergard Animal Cage Changing Station

35. The downdraft airflow pattern provides personnel allergen protection from animal hair, dander, and bedding materials HEPA filtered Protection provided to animals, cages and bedding The downdraft airflow pattern provides personnel allergen protection from animal hair, dander, and bedding materials HEPA filtered Protection provided to animals, cages and bedding Cage Changing Station Features

36. Personnel working with station should wear personal protective equipment Any required vivarium clothing Lab coat/wrap around gown Gloves Safety glasses Decontaminate surface with an appropriate disinfectant at the end of each work operation Personnel working with station should wear personal protective equipment Any required vivarium clothing Lab coat/wrap around gown Gloves Safety glasses Decontaminate surface with an appropriate disinfectant at the end of each work operation Safe Use of a Animal Changing Station

37. Laminar Air Flow Workstation

38. Provides the ideal particle free, bacteria free, clean air environment HEPA filtered Positive Air Flow-outflow towards worker Creates airflow throughout the entire work zone Protects animals, not personnel Not for use with biohazardous animals!!!! Provides the ideal particle free, bacteria free, clean air environment HEPA filtered Positive Air Flow-outflow towards worker Creates airflow throughout the entire work zone Protects animals, not personnel Not for use with biohazardous animals!!!! Laminar Air Hood Features

39.  Personnel working with station should wear personal protective equipment ◦ Any required vivarium clothing ◦ Lab coat/wrap around gown ◦ Gloves ◦ Safety glasses  Decontaminate with an appropriate disinfectant at the end of each work operation  Not for use with biohazardous animals!!  Personnel working with station should wear personal protective equipment ◦ Any required vivarium clothing ◦ Lab coat/wrap around gown ◦ Gloves ◦ Safety glasses  Decontaminate with an appropriate disinfectant at the end of each work operation  Not for use with biohazardous animals!! Safe Use of Laminar Flow Hood

40. Questions?