2Introduction to Biological Safety Biological Hazards are divided into 4 Biosafety LevelsBSL 1BSL 2BSL 3BSL4Biosafety levels define the lab requirements, protective clothing, and work practices
3BioSafety Definitions BSL 1 (BMBL)Not known to consequently cause disease in healthy human adultsPose minimal hazards under ordinary conditions of handling
4BioSafety Level 1 Standard Practices Use Mechanical Pipetting devices No Eating, Drinking, Smoking in LabMinimize splashes and aerosolsDecontaminate work surfacesSafe handling of sharpsWash Hands before leaving lab
6BioSafety Definitions BSL 2 (BMBL)suitable for work involving well-characterized agents not known to cause disease in healthy adult humans, and of minimal potential hazard to laboratory personnel and the environmentExamples: Measles Virus, Salmonella,Hepatitis B Virus
7BioSafety Definitions Other Materials Handled at BSL 2(Requires compliance with OSHA BBP Standard)Human BloodHuman Blood ComponentsHuman TissuesHuman Cell Lines
8BioSafety Level 2 Standard Practices All requirements for BSL 1 plus: Access to laboratory is limited or restricted when work is being conductedPersonnel have specific training in handling pathogenic agentsBiohazard Sign posted on the doorExtreme precautions are taken with contaminated sharp items
9BioSafety Level 2 Standard Practices cont. Written Biosafety ProceduresReport SpillsMinimize aerosol generationPersonnel receive appropriate immunizations or test (e.g. Hep B vaccine or TB skin Testing)
10BioSafety Level 2 Written Biosafety Procedures Prepared specifically for lab by PIIncorporated into Lab Safety PlanPersonnel are advised of special hazardsPersonnel are required to read and follow procedures
11BioSafety Level 2 Personal Protective Equipment Lab Coats Gloves Double Gloving when appropriateEye Protection - Safety Glasses/Shield
12BioSafety Level 3 BSL 3 (BMBL) applicable 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 as a result of exposure by the inhalation route.
13BioSafety Level 3 Standard Practices All requirements for BSL 2 plus: Limited lab access2 doors in a series to access labAble to wash entire labSpecial exhaust ventilation (Not re circulated, negative pressure)
14BioSafety Level 3 Standard Practices –cont. High level of training Personnel receive vaccinations if availableWork in Biosafety cabinets
16BioSafety Level 4 BSL 4 (BMBL) required for work with dangerous and exotic agents which pose a high individual risk of aerosol-transmitted laboratory infections and life-threatening disease.
17BioSafety Level 4 Requirements All requirements for BSL3 plus: Class III Biosafety cabinet or positive pressure suitsShower/Change roomsClothing Autoclaved before launderingAir Locks
18BioSafety Level 4ExamplesEbola VirusMonkey B VirusMarburg Virus
19ContainmentPrimaryProtection of personnel and immediate lab from exposureGood TechniquesSafety equipmentBiological Safety CabinetEngineering ControlsPPESerologic surveillance
20Containment Secondary Protection of environment external to lab Facility DesignOperational practices
21Safe Work Practices for all Levels Wash hands after work; when removing gloves; before leaving labNo eating, drinking, applying cosmetics, handling contact lenses in the lab.Maintain labs in clean, orderly fashion.
22Safe Work Practices for all Levels Limit access to lab when work with organisms is in progressUse good microbiological techniquesNo mouth pipettingWhen possible use plastic instead of glass
23Sharps SafetySharps include needles, syringes, razor blades, lancets, slides, scalpels, pipettes, micropipettes, pipette tips, broken plastic or glassware, and other devices capable of cutting or piercing the skin.
24Sharps SafetyContaminated needles shall not be bent, recapped, or removed unless there is no feasible alternative.If required, use a mechanical device or a one handed technique.
25Sharps SafetySafety devices or alternatives to needles should be used when available.
26Sharps DisposalSharps containers for disposal of these items should be conveniently located and easily accessible in all work places where sharps are used.
27Sharps DisposalSyringes and syringes without a needle attached go into a sharps containerContaminated micropipettes, pipette tips, and Pasteur pipettes are discarded in a puncture-resistant container or a sharps container for disposal.
28Sharps DisposalDon’t place needles or sharps in office waste containers
29Laminar Flow Equipment BioSafety Cabinets (BSCs)contain infectious agents to protect personnel and the environmentLaminar Flow Clean Benches (LFBs)Non Hazardous work onlyProtect work from contamination
30BioSafety Cabinets 3 Classes All exhaust is HEPA filtered before leaving the cabinetClass IDo not protect the work from contaminationAir entering cabinet is not filtered
31Biosafety Cabinets Class II (4 types – A, B1, B2, and B3) Each type recirculates different amount of airSome are hard ducted, and some exhaust into the room.Class IIITotally enclosed, ventilated cabinetsWork through portals with attached gloves
32BioSafety CabinetsAll equipment is laid out to not restrict airflow in the cabinet
33Laminar Flow Benches Do not protect personnel or the environment Discharges HEPA filtered air across the work surface toward the user.
34Use of Laminar Flow Equipment Minimize airflow disturbances (moving in/out of cabinet, people walking by, opening doors, blocking grills with equipment)Wipe down surfaces with approved disinfectant before and after work.
35Use of Laminar Flow Equipment Minimize use of flames in cabinetTry not to use chemicals in Class I or II hoods that are ductedDo not store excess equipment in cabinetAll laminar flow equipment must be certified annually.
38Centrifuge Operation Check tubes for cracks/chips Use matched sets of tubes, buckets, etc. Make sure the rotor is balanced properlyTightly seal all tubes and safety cupsEnsure that rotor is locked to spindle and the bucket is seatedClose lid during operationAllow to come to complete stop before opening
39Centrifuge SafetyUse sealed tubes, safety buckets or rotors when possibleWhen possible fill and open centrifuge tubes or buckets in the biological safety cabinet.Lubricate O-rings and rotor threads weeklyDisinfect weekly and after all spills or breaks
40Biohazardous WasteAll cultures, stocks, and other regulated waste are decontaminated before disposal by an approved decontamination method such as autoclaving.
41Biohazardous Waste Examples stock cultures materials containing or contaminated with bloodblood and body fluidssharpspipette tipsanimal wasteother contaminated lab material
42Blood and Body Fluids Waste Serum,PlasmaOther blood componentsSemenVaginal secretionsCerebrospinal fluidPleural fluidPeritoneal fluidPericardial fluidAmniotic fluidAny other body fluid visibly contaminated with blood