2 Hierarchy of Controls Administrative Control Engineering Control Work PracticesPersonal Protective Clothing or Equipment
3 Administrative Controls SubstitutionAuthorization/ApprovalWritten biosafety procedures required for the experimental procedures and equipment including inventory of biological agents or materialsLaboratory personnel biosafety trainingMedical Surveillance (BSL 2 and above)Health historyMedical screeningImmunizationSerum storagePost-exposure prophylaxis
5 Personal Protective Clothing and Equipment Provides barrier against skin, mucous membrane or respiratory exposure to infectious agents during proceduresPrevent spread of contaminationDoes not eliminate the hazardIntegrity wanes with use (i.e., change gloves frequently)
6 BSL 1: Work Practices and Procedures ApplicationsNon-infectious agent and tissue culture, media preparationPrevent Cross ContaminationKeep cultures coveredFlame instruments and containersUse sterile media and equipmentKeep hands or face away from cultures
7 BSL 1: Work Practices and Procedures Biosafety ProceduresWork with agents may be conducted on open benchWash hands oftenNo mouth pipettingNo eating or drinking in labMinimize aerosol generationDecontaminate work surfacesWear applicable PPE
8 BSL 2: Work Practices and Procedures Increasing emphasis on safety procedures and practicesIncreasing need for staff trainingIncreasing need for competent supervisionBiohazard sign posted at entry doorBiohazard labels affixed on regulated waste containersUse of personal protective equipment as a barrier to exposure: lab coat, gloves, eye and face protectionSome work on open bench allowed
9 BSL 2: Work Practices and Procedures Aerosol generating procedures performed in a biosafety cabinet:HomogenizingVortexingVigorous mixingPipetting infectious liquidsSonicationPouringIf breach occurs:Evacuate lab, post spill signWith appropriate PPE and disinfectant, decontaminate centrifuge, buckets, other items or areas
10 Correct Use of Biosafety Cabinets Start UpTurn off ultraviolet light (if so equipped) as soon as you enter the room.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).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 UseLoad supplies prior to work.Do not overload cabinet.Separate clean and dirty side.Work in center of work area.Do not block front or rear grills.Minimize disruption of airflow (turbulence).Clean up spill promptly.Discard waste within the cabinet.
12 Correct Use of Biosafety Cabinets Shut DownDecontaminate and remove all items from interior work area.Decontaminate readily accessible interior surfaces with a disinfectant appropriate for the agents or suspected agents present.Turn on ultraviolet light (if so equipped).Allow five minutes of operation to purge system. Then wait at least minutes.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.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.
14 Correct Use of Biosafety Cabinets – Open Flames DO NOT use Bunsen burners or open flamesFire hazardCan damage HEPA filterInterferes with proper air flowMicroincinerator preferredBurner with pilot light not a good alternativeOpen flames react with disinfectants (flammables)
15 Safe Use of CentrifugeUse sealed tubes, rotors, and safety cups/buckets that are sealed with O-ringsInspect tubes, O-rings and rotors for wear, and buckets for cracks, chips, erosion, etc.Do not use aluminum foil to cap centrifuge tubesClean and maintain gaskets and O-ringsChange O-rings if compromisedLoad/unload centrifuge tubes, rotors and accessories in BSCSmall, low speed centrifuges may be placed in a BSC; however, high speed centrifuges pose additional hazardsDo not overfill tubesBalance buckets, tubes and rotors properly before centrifugationWait 5 minutes (or 30 mins. for high speed centrifuge) after each run before openingDo not decant or pour off supernatant. Use a vacuum system with appropriate in-line reservoirs and filters
16 Safe Use of BlendersAvoid use of glass blender jars, unless covered with polypropylene jarPlace disinfectant-moistened towel over the top of the blender during useBefore opening the blender jar, allow the unit to rest at least 1 minute for aerosols to settle and then open in a BSCDecontaminate promptly after use
17 Minimizing Aerosols Use careful pipetting practices Avoid drops onto hard surfacesWipe up spills promptly with appropriate disinfectantFor ejection of liquid from micropipetteNo blowoutNo pressure ejectionUse wall contactUse capped tubes when mixing, blending, or vortexingPour liquids carefullyAvoid bubbles
18 Careful Pipetting Techniques Never blow out last drop in pipetteUse pipetting aids with filtersNever mix by suction and expulsion (mix by sonication)Discharge liquid down side of container, using tip-to-wall contactDeliver as close as possible to contentsWork over plastic-backed absorbent matting (ensure it doesn’t slide forward or backward blocking air grill)
19 Use Extreme Care with Sharps Use sharps if only absolutely required as part of a processPercutaneous exposure riskEmploy safe work practicesUtilize safe sharp devicesAerosol exposure riskUse biosafety cabinet for removal of air from needleUse mechanical methods for needle removalNever bend, recap or manipulate sharps by handKeep hands away from needle
20 Vacuum System Protection In-line filter and disinfectant in collection and overflow flasks