Biological Safety Training: BSL Levels & Lab Practices
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Biological Safety Training East Carolina University Office of Prospective Health/Biological Safety Introduction to Biological Safety Biological Hazards are divided into 4 Biosafety Levels – – – – BSL 1 BSL 2 BSL 3 BSL4 Biosafety levels define the lab requirements, protective clothing, and work practices East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm BioSafety Definitions BSL 1 (BMBL) – Not known to cause disease in healthy human adults – Pose minimal hazards under ordinary conditions of handling East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm BioSafety Level 1 • Standard Practices – Use Mechanical Pipetting devices – No Eating, Drinking, Smoking in Lab – Minimize splashes and aerosols – Decontaminate work surfaces – Safe handling of sharps – Wash Hands before leaving lab – Biohazard sign is posted East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm BioSafety Level 1 • Protective Clothing – Lab Coat – Gloves – Eye Protection (if splashes are likely) East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm BioSafety Definitions • BSL 2 (BMBL) – Work with well-characterized agents not known to cause disease in healthy adult humans; minimal hazard to laboratory personnel and the environment Examples: Salmonella, Hepatitis B Virus Measles Virus (not aerosolized) East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm BioSafety Definitions • Other Materials Handled at BSL 2 (Requires compliance with OSHA BBP Standard) – – – – – Human Blood Human Tissues Human Cell Lines Biotoxins Viral Vectors East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm BioSafety Level 2 • Standard Practices – All requirements for BSL 1 plus: • Access to laboratory is limited or restricted when work is being conducted; door is closed • Personnel have specific training in handling pathogenic agents • Extreme precautions are taken with contaminated sharp items East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm BioSafety Level 2 • Standard Practices cont. • • • • Written Biosafety Procedures Report Spills Minimize aerosol generation Personnel receive immunizations or testing (e.g. Hep B vaccine or TB skin Testing) East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm BioSafety Level 2 • Personal Protective Equipment – Lab Coats – Gloves • Double Gloving when appropriate – Eye Protection - Safety Glasses/Shield – Biosafety Cabinets or sealed safety centrifuge East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm BioSafety Level 3 • BSL 3 (BMBL) – Use of indigenous or exotic agents which may cause serious or potentially lethal disease from exposure by the inhalation route. East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm BioSafety Level 3 • Standard Practices – All requirements for BSL 2 plus: • • • • Very limited lab access 2 doors in a series to access lab Able to decontaminate entire lab Special exhaust ventilation (Not re-circulated, no airflow outward) East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm BioSafety Level 3 • Standard Practices –cont. – High level of training – Personnel receive vaccinations if available – Work in Biosafety cabinets and/or respirator used East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm BioSafety Level 3 • Examples – Mycobacterium Tuberculosis – Hanta virus – SARS East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm BioSafety Level 4 • BSL 4 (BMBL) – dangerous and exotic agents with a high risk of aerosol-transmitted laboratory infections and life-threatening disease. East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm BioSafety Level 4 • Requirements – All requirements for BSL3 plus: • Class III Biosafety cabinet or positive pressure suits • Shower/Change rooms • Clothing Autoclaved before laundering • Air Locks • All waste contained and decontaminated onsite • Space Suits East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm Space Suit East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm BioSafety Level 4 • Examples – Ebola Virus – Monkey B Virus – Marburg Virus East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm Safe Work Practices for all Levels • Wash hands after work; when removing gloves; before leaving lab • No eating, drinking, applying cosmetics, handling contact lenses in the lab. • Maintain labs in clean, orderly fashion. East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm Safe Work Practices for all Levels • Limit access to lab when work with organisms is in progress • Use good microbiological techniques • No mouth pipetting • When possible use plastic instead of glass East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm Sharps Safety • Sharps 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. East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm Sharps Safety • Contaminated needles are not bent, recapped, or removed. • If recapping is required, use a mechanical device or a one handed technique. East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm Sharps Safety • Safety devices or alternatives to needles should be used when available. East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm Sharps Disposal • Sharps containers for disposal of these items are conveniently located and easily accessible. East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm Sharps Disposal • Syringes and syringes without a needle attached go into a sharps container • Contaminated micropipettes, pipette tips, and Pasteur pipettes are discarded in a puncture-resistant container or a sharps container for disposal. East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm Sharps Disposal • Don’t place needles or sharps in soft-sided bags. East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm Laminar Flow Equipment BioSafety Cabinets (BSCs) Protect personnel and the environment from hazardous particulates and from infectious agents Laminar Flow Clean Benches (LFBs) Non Hazardous work only Protect work from contamination *Do not protect personnel* East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm BioSafety Cabinets • 3 Classes • All exhaust is HEPA filtered before leaving the cabinet East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm NIH Guidelines for Use of Recombinant DNA apply to all work at ECU, not just NIH-funded work. The university can be sanctioned for failure to comply. East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm NIH Guidelines may apply to work with: • • • • • • Recombinant DNA or RNA Genetic/Recombinant Techniques, molecules or organisms Transgenic plants or animals or knock-out organisms Viral or host vector systems or plasmids Human gene transfer Transfer of drug-resistance to microorganisms not known to acquire it naturally • Formation of DNA or RNA genes to synthesize toxin molecules East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm At ECU, all work which may be subject to the NIH Guidelines must be reviewed by the Biological Safety to determine whether the work is “NIH exempt.” The researcher may not make this determination independently East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm Submission of a Biological Safety registration to the Institutional Biological Safety Committee will ensure that full review per the NIH Guidelines occurs. Form is available on the Prospective Health website http://www.ecu.edu/csdhs/prospectivehealth/Biological-Safety-Office-ofProspective-Health.cfm East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm Some work must be approved by NIH as well as by the local Institutional Biological Safety Committee: • Transfer of Drug Resistance to microorganisms not known to acquire naturally • Formation of genes to synthesize toxins lethal to vertebrates at an LD50 of < 100 nanograms per kg body weight (e.g., tetanus toxin) • Human gene transfer East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm Drug resistance is of concern if it could compromise the use of the drug to control disease in humans, veterinary medicine or agriculture. Only NIH may determine whether the drug resistance can compromise therapeutic use! This is not a local or researcher decision. East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm Biological Safety interfaces with the Human Subjects Institutional Review Board (IRB) and Institutional Animal Care and Use Committee (IACUC) to ensure that the NIH Guidelines are met at ECU. East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm The IBC must report significant violations of the NIH Guidelines, as well as researchrelated accidents and illnesses to the NIH Office of Biotechnology Affairs. East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm Contact Biological Safety, Office of Prospective Health for questions. East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm For more information see the ECU Biological Safety Manual Office of Prospective Health/Biological Safety Biological Safety Committee East Carolina University http://www.ecu.edu/cs-dhs/prospectivehealth/customcf/biosafety/BIOLOGICAL_SAFETY_MANUAL%20_WEB.pdf East Carolina University Office of Prospective Health/Biological Safety http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
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