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Biosafety Practices and Procedures. Hierarchy of Controls Administrative Control Administrative Control Engineering Control Engineering Control Work Practices.

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Presentation on theme: "Biosafety Practices and Procedures. Hierarchy of Controls Administrative Control Administrative Control Engineering Control Engineering Control Work Practices."— Presentation transcript:

1 Biosafety Practices and Procedures

2 Hierarchy of Controls Administrative Control Administrative Control Engineering Control Engineering Control Work Practices Work Practices Personal Protective Clothing or Equipment Personal Protective Clothing or Equipment

3 Administrative Controls Substitution Substitution Authorization/Approval Authorization/Approval Written biosafety procedures required for the experimental procedures and equipment including inventory of biological agents or materials Written biosafety procedures required for the experimental procedures and equipment including inventory of biological agents or materials Laboratory personnel biosafety training Laboratory personnel biosafety training Medical Surveillance (BSL 2 and above) Medical Surveillance (BSL 2 and above) Health history Health history Medical screening Medical screening Immunization Immunization Serum storage Serum storage Post-exposure prophylaxis Post-exposure prophylaxis

4 Engineering Controls Biological safety cabinets, glove boxes Biological safety cabinets, glove boxes Animal containment caging systems Animal containment caging systems Safety equipment (filtered or sealed equipment) Safety equipment (filtered or sealed equipment) Ventilation system Ventilation system Containment facilities Containment facilities

5 Personal Protective Clothing and Equipment Provides barrier against skin, mucous membrane or respiratory exposure to infectious agents during procedures Provides barrier against skin, mucous membrane or respiratory exposure to infectious agents during procedures Prevent spread of contamination Prevent spread of contamination Does not eliminate the hazard Does not eliminate the hazard Integrity wanes with use (i.e., change gloves frequently) Integrity wanes with use (i.e., change gloves frequently)

6 BSL 1: Work Practices and Procedures Applications Applications Non-infectious agent and tissue culture, media preparation Non-infectious agent and tissue culture, media preparation Prevent Cross Contamination Prevent Cross Contamination Keep cultures covered Keep cultures covered Flame instruments and containers Flame instruments and containers Use sterile media and equipment Use sterile media and equipment Keep hands or face away from cultures Keep hands or face away from cultures

7 Biosafety Procedures Biosafety Procedures Work with agents may be conducted on open bench Work with agents may be conducted on open bench Wash hands often Wash hands often No mouth pipetting No mouth pipetting No eating or drinking in lab No eating or drinking in lab Minimize aerosol generation Minimize aerosol generation Decontaminate work surfaces Decontaminate work surfaces Wear applicable PPE Wear applicable PPE BSL 1: Work Practices and Procedures

8 BSL 2: Work Practices and Procedures Increasing emphasis on safety procedures and practices Increasing emphasis on safety procedures and practices Increasing need for staff training Increasing need for staff training Increasing need for competent supervision Increasing need for competent supervision Biohazard sign posted at entry door Biohazard sign posted at entry door Biohazard labels affixed on regulated waste containers Biohazard labels affixed on regulated waste containers Use of personal protective equipment as a barrier to exposure: lab coat, gloves, eye and face protection Use of personal protective equipment as a barrier to exposure: lab coat, gloves, eye and face protection Some work on open bench allowed Some work on open bench allowed

9 BSL 2: Work Practices and Procedures Aerosol generating procedures performed in a biosafety cabinet: Aerosol generating procedures performed in a biosafety cabinet: Homogenizing Homogenizing Vortexing Vortexing Vigorous mixing Vigorous mixing Pipetting infectious liquids Pipetting infectious liquids Sonication Sonication Pouring Pouring If breach occurs: If breach occurs: Evacuate lab, post spill sign Evacuate lab, post spill sign With appropriate PPE and disinfectant, decontaminate centrifuge, buckets, other items or areas With appropriate PPE and disinfectant, decontaminate centrifuge, buckets, other items or areas

10 Correct Use of Biosafety Cabinets Start Up Start Up Turn off ultraviolet light (if so equipped) as soon as you enter the room. Turn off ultraviolet light (if so equipped) as soon as you enter the room. Turn on all blowers and BSC illumination lights. Turn on all blowers and BSC illumination lights. Allow five minutes of operation to purge system; check flow alarm system audio and visual alarm function (if so equipped). Allow five minutes of operation to purge system; check flow alarm system audio and visual alarm function (if so equipped). Decontaminate readily accessible interior surfaces and items with a disinfectant (appropriate for the agents or suspected agents present) before loading and wait at least 10 minutes prior to start of work. Decontaminate readily accessible interior surfaces and items with a disinfectant (appropriate for the agents or suspected agents present) before loading and wait at least 10 minutes prior to start of work.

11 Correct Use of Biosafety Cabinets During Use During Use Load supplies prior to work. Load supplies prior to work. Do not overload cabinet. Do not overload cabinet. Separate clean and dirty side. Separate clean and dirty side. Work in center of work area. Work in center of work area. Do not block front or rear grills. Do not block front or rear grills. Minimize disruption of airflow (turbulence). Minimize disruption of airflow (turbulence). Clean up spill promptly. Clean up spill promptly. Discard waste within the cabinet. Discard waste within the cabinet.

12 Correct Use of Biosafety Cabinets Shut Down Shut Down Decontaminate and remove all items from interior work area. Decontaminate and remove all items from interior work area. Decontaminate readily accessible interior surfaces with a disinfectant appropriate for the agents or suspected agents present. Decontaminate readily accessible interior surfaces with a disinfectant appropriate for the agents or suspected agents present. Turn on ultraviolet light (if so equipped). Turn on ultraviolet light (if so equipped). Allow five minutes of operation to purge system. Then wait at least 10 minutes. Allow five minutes of operation to purge system. Then wait at least 10 minutes. Turn off BSC blower. Turn off BSC blower.

13 Correct Use of Biosafety Cabinets Moving/Installation BSCs must be decontaminated prior to moving. In order to ensure filter integrity, the equipment must be recertified after the BSC is installed at its final new location. Arrangements need to be made well in advance in order for contractors to meet your schedule. The PI is responsible for contacting the contractor or to schedule this work. Moving/Installation BSCs must be decontaminated prior to moving. In order to ensure filter integrity, the equipment must be recertified after the BSC is installed at its final new location. Arrangements need to be made well in advance in order for contractors to meet your schedule. The PI is responsible for contacting the contractor or to schedule this work. Decontamination Decontamination is usually performed by certified professionals. Decontamination Decontamination is usually performed by certified professionals. Certification All BSCs that are used for handling biohazardous materials must be recertified annually. SDSU has contracted with a specific contractor to provide a consistent level of certification and maintenance service. Contact EHS at (619) to obtain contractor information. Certification All BSCs that are used for handling biohazardous materials must be recertified annually. SDSU has contracted with a specific contractor to provide a consistent level of certification and maintenance service. Contact EHS at (619) to obtain contractor information.

14 DO NOT use Bunsen burners or open flames DO NOT use Bunsen burners or open flames Fire hazard Fire hazard Can damage HEPA filter Can damage HEPA filter Interferes with proper air flow Interferes with proper air flow Microincinerator preferred Microincinerator preferred Burner with pilot light not a good alternative Burner with pilot light not a good alternative Open flames react with disinfectants (flammables) Open flames react with disinfectants (flammables) Correct Use of Biosafety Cabinets – Open Flames

15 Use sealed tubes, rotors, and safety cups/buckets that are sealed with O-rings Use sealed tubes, rotors, and safety cups/buckets that are sealed with O-rings Inspect tubes, O-rings and rotors for wear, and buckets for cracks, chips, erosion, etc. Inspect tubes, O-rings and rotors for wear, and buckets for cracks, chips, erosion, etc. Do not use aluminum foil to cap centrifuge tubes Do not use aluminum foil to cap centrifuge tubes Clean and maintain gaskets and O-rings Clean and maintain gaskets and O-rings Change O-rings if compromised Change O-rings if compromised Load/unload centrifuge tubes, rotors and accessories in BSC Load/unload centrifuge tubes, rotors and accessories in BSC Small, low speed centrifuges may be placed in a BSC; however, high speed centrifuges pose additional hazards Small, low speed centrifuges may be placed in a BSC; however, high speed centrifuges pose additional hazards Do not overfill tubes Do not overfill tubes Balance buckets, tubes and rotors properly before centrifugation Balance buckets, tubes and rotors properly before centrifugation Wait 5 minutes (or 30 mins. for high speed centrifuge) after each run before opening Wait 5 minutes (or 30 mins. for high speed centrifuge) after each run before opening Do not decant or pour off supernatant. Use a vacuum system with appropriate in-line reservoirs and filters Do not decant or pour off supernatant. Use a vacuum system with appropriate in-line reservoirs and filters Safe Use of Centrifuge

16 Avoid use of glass blender jars, unless covered with polypropylene jar Avoid use of glass blender jars, unless covered with polypropylene jar Place disinfectant-moistened towel over the top of the blender during use Place disinfectant-moistened towel over the top of the blender during use Before opening the blender jar, allow the unit to rest at least 1 minute for aerosols to settle and then open in a BSC Before opening the blender jar, allow the unit to rest at least 1 minute for aerosols to settle and then open in a BSC Decontaminate promptly after use Decontaminate promptly after use Safe Use of Blenders

17 Use careful pipetting practices Use careful pipetting practices Avoid drops onto hard surfaces Avoid drops onto hard surfaces Wipe up spills promptly with appropriate disinfectant Wipe up spills promptly with appropriate disinfectant For ejection of liquid from micropipette For ejection of liquid from micropipette No blowout No blowout No pressure ejection No pressure ejection Use wall contact Use wall contact Use capped tubes when mixing, blending, or vortexing Use capped tubes when mixing, blending, or vortexing Pour liquids carefully Pour liquids carefully Avoid bubbles Avoid bubbles Minimizing Aerosols

18 Careful Pipetting Techniques Never blow out last drop in pipette Never blow out last drop in pipette Use pipetting aids with filters Use pipetting aids with filters Never mix by suction and expulsion (mix by sonication) Never mix by suction and expulsion (mix by sonication) Discharge liquid down side of container, using tip-to-wall contact Discharge liquid down side of container, using tip-to-wall contact Deliver as close as possible to contents Deliver as close as possible to contents Work over plastic-backed absorbent matting (ensure it doesnt slide forward or backward blocking air grill) Work over plastic-backed absorbent matting (ensure it doesnt slide forward or backward blocking air grill)

19 Use Extreme Care with Sharps Use sharps if only absolutely required as part of a process Use sharps if only absolutely required as part of a process Percutaneous exposure risk Percutaneous exposure risk Employ safe work practices Employ safe work practices Utilize safe sharp devices Utilize safe sharp devices Aerosol exposure risk Aerosol exposure risk Use biosafety cabinet for removal of air from needle Use biosafety cabinet for removal of air from needle Use mechanical methods for needle removal Use mechanical methods for needle removal Never bend, recap or manipulate sharps by hand Never bend, recap or manipulate sharps by hand Keep hands away from needle Keep hands away from needle

20 Vacuum System Protection In-line filter and disinfectant in collection and overflow flasks In-line filter and disinfectant in collection and overflow flasks


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