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1、BiosafetyPractices and ProceduresPresented by:Yang Y19 Jun 2012BiosafetyConcept:Biological safety is to prevent occupational acquired infections as well as the release of pathogens into the environment.Biosafety is the prevention of large-scale loss of biological integrity,focusing both on ecology a
2、nd human health.Biological&BiosecurityBiosafety is related to several fieldsIn ecologyIn agricultureIn medicineIn chemistryIn exobiologyPrecautionary principle Biosafety require application of the precautionary principle in order to prevent the accidental release of potentially destructive biologica
3、l material.Biosafety levelA biosafety level is the level of the biocontainment precautions required to isolate dangerous biological agents in an enclosed facility.Biocontainment can be classified by the relative danger to the surrounding environment as biological safety levels(BSL).The levels of con
4、tainment range from the lowest biosafety level 1(BSL-1)to the highest at level 4(BSL-4).Biosafety level 1This level is suitable for work involving well-characterized agents not known to consistently cause disease in healthy adult humans,and of minimal potential hazard to laboratory personnel and the
5、 environment.Such as canine hepatitis,non-pathogenic Escherichia coli,as well as some cell cultures and non-infectious bacteria.Biosafety level 2This level is similar to Biosafety Level 1 and is suitable for work involving agents of moderate potential hazard to personnel and the environment.It inclu
6、des various bacteria and viruses that cause only mild disease to humans,or are difficult to contract via aerosol in a lab setting.Such as C.difficile,most Chlamydiae,hepatitis A,B,and C,influenza A,Lyme disease,Salmonella,mumps,measles,HIV,scrapie,MRSA,and VRSA.Biosafety level 3This level is applica
7、ble to clinical,diagnostic,teaching,research,or production facilities in which work is done with indigenous or exotic agents which may cause serious or potentially lethal disease after inhalation.It includes various bacteria,parasites and viruses that can cause severe to fatal disease in humans but
8、for which treatments exist.Such as Mycobacterium tuberculosis,Bacillus anthracis,SARS coronavirus,Salmonella typhi,and yellow fever virus.Biosafety level 4This level is required for work with dangerous and exotic agents that pose a high individual risk of aerosol-transmitted laboratory infections,ag
9、ents which cause severe to fatal disease in humans for which vaccines or other treatments are not available.Such as Ebola virus,Lassa feverBSL-2 differs from BSL-1 in thatLaboratory personnel have specific training in handling pathogenic;Access to the laboratory is limited when work is being conduct
10、ed;Extreme precautions are taken with contaminated sharp items;Certain procedures in which infectious aerosols or splashes may be created are conducted in biological safety cabinets or other physical containment equipment.AerosolAn aerosol is a colloid suspension of fine solid particles or liquid dr
11、oplets in a gas.Such as smog and smokeAerosols in the Laboratory bottles and flasks;blenders and vortexers;centrifuges;homogenizers;needles and syringes;pipettes;pressurized vessels;rubber stoppers;shakers;sonicators;vacuum and aspirating equipmentBiosafety cabinet A biosafety cabinet(BSC)is an encl
12、osed,ventilated laboratory workspace for safely working with materials contaminated with pathogens requiring a defined biosafety level.Purposes of Biosafety cabinetThe primary purpose of a BSC is to serve as the primary means to protect the laboratory worker and the surrounding environment from path
13、ogens.All exhaust air is HEPA-filtered as it exits the biosafety cabinet,removing harmful bacteria and viruses.Classes of Biosafety cabinetClass II,Type A1,A2,B1,B2 CabinetAir flow in Class II,Type A2Correct Use of Biosafety CabinetsStart UpTurn off ultraviolet light(if so equipped)as soon as you en
14、ter 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 suspecte
15、d agents present)before loading and wait at least 10 minutes prior to start of work.Correct Use of Biosafety CabinetsDuring Use Load 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 airfl
16、ow(turbulence).Clean up spill promptly.Discard waste within the cabinet.Correct Use of Biosafety CabinetsShut 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.T
17、urn on ultraviolet light(if so equipped).Allow five minutes of operation to purge system.Then wait at least 10 minutes.Never turn off BSC blower.DO NOT open flamesFire hazardCan damage HEPA filterInterferes with proper air flowOpen flames react with disinfectants(flammables)Correct Use of Biosafety
18、Cabinets Open FlamesAvoid 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 promptl
19、y after useSafe Use of BlendersUse careful pipetting practicesAvoid 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 carefu
20、llyAvoid bubblesMinimizing AerosolsCareful Pipetting TechniquesNever 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 as possible to contentsWork over plastic-b
21、acked absorbent matting(ensure it doesnt slide forward or backward blocking air grill)Use Extreme Care with SharpsUse 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 needleBiological Waste DisposalPuncture-proof,leak-proof,sealable receptaclesAvoid over-fillingDispose properlyTo be continued
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