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Biosafety in Research Laboratories

Biosafety in Research Laboratories. Definitions. Biohazard An agent of biological origin that has the capacity to produce negative effects on humans, plants or animals. Biosafety

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Biosafety in Research Laboratories

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  1. Biosafety in Research Laboratories

  2. Definitions Biohazard An agent of biological origin that has the capacity to produce negative effects on humans, plants or animals. Biosafety The application of: lab practices and procedures, specific lab facilities and safety equipment to protect against exposure to potentially infectious material

  3. What is Biohazardous Material? • Bacteria • Viruses • Parasites • Fungi • Human and Non-Human Primate Material • Recombinant DNA • Animals • Biological Toxins

  4. How Can You Be Exposed? • Inhalation -80% of lab acquired infections (LAI) • Perform experiments in biological safety cabinets • Use respiratory protection Remaining 20% LAI are a combination of: • Percutaneous • Avoid sharps when possible, use plastic pipettes and disposable glassware • Mucous Membrane (eyes, nose and mouth) • Wash hands often, avoid touching face • Ingestion • Wash hands often, No eating, drinking, smoking or applying cosmetics • All exposures must be reported to the PI, Occ. Health, Biosafety

  5. How Can You Protect Yourself? • Administrative Controls • Use attenuated or non-virulent strains, require restraint during animal injection, avoid sharps and glassware, require decontamination, Standard Operating Procedures, training • Engineering Controls • Biosafety Cabinets, Sealed Rotors/Safety Cups, Pipette Aids, Safety Enclosures for Cell Sorters • Personal Protective Equipment • Lab Coat / gown, gloves, goggles, facial protection • Close toed shoes

  6. Guidelines and Regulations • Biosafety in Microbiological and Biomedical Laboratories (CDC) • NIH Guidelines for Research Involving Recombinant DNA Molecules (NIH) • OSHA Bloodborne Pathogens Standard (OSHA) • Committee on Microbiological Safety (COMS)

  7. Why Biosafety Guidelines? • Protection • Self, Co-workers, Lab support personnel • Emergency Responders • Experiment and Materials • Environment • Compliance with NIH, CDC, OSHA, City of Boston, and Town of Belmont

  8. Safety Equipment(Primary Barriers) • Biosafety cabinets (BSCs) • Safety centrifuge cups and rotors • Pipetting Devices • Vacuum line HEPA Filters • Personal protective clothing • Gloves (remember the one glove rule) • Gowns/Lab coats (should remain inside the lab) • Eye and face protection • Close toed shoes

  9. Biological Safety Cabinets Must be Certified Annually • Air Systems Technologies (508) 427 5558 BSC Provide: • Product Protection • Personnel Protection • Environmental Protection Use biosafety cabinets (class II) for work with infectious agents involving: • Aerosols and splashes • Large volumes, High concentrations • BL2 materials

  10. Biological Safety Cabinets • Protection against particulates only • No gas or vapor protection • Air flow is easily disrupted • Don’t cover the grill • Move arms slowly in and out of cabinet • Walk slowly • 70% return 30% exhausted

  11. Centrifuges • Safety Cups and Rotors • Load and unload safety cups inside the BSC

  12. Vacuum Line HEPA Filter Protection • Placed in between the vacuum system and the aspiration flasks

  13. Biosafety Levels • BSL-1: agents not known to cause disease (in healthy adults) • BSL-2: agents associated with disease • BSL-3: indigenous/exotic agents associated with human disease and with potential for aerosol transmission • BSL-4: dangerous/exotic agents of life threatening nature

  14. Biosafety Levels • BSL-2S (with Stipulation) • Harvard classification not recognized nationally • Work can be done inside a BL2 structural facility while practicing certain BL3 practices and procedures • disposable sleeve covers or gowns,..... • Stipulated by the Institutional Biosafety Committee (COMS)

  15. Biosafety Level 1 • Immunocompromised workers are at risk • Standard Operating Procedures • Minimize splashes and aerosols • PPE • Decontaminate work surfaces daily • Decontaminate wastes • Maintain insect & rodent control program • Examples: • Adeno associated virus • Murine leukemia virus • Non pathogenic E. coli • Animal tissue and cells (other than NHP)

  16. Biosafety Level 1Laboratory Facilities (Secondary Barriers)

  17. Biosafety Level 2 • Infectious dose will vary based on: • Agent • Exposure time • Exposure route • Immunization or antibiotic treatment may be available • Extreme precaution with contaminated needles or sharp instruments • Examples: • Hepatitis B Virus • Influenza • Salmonella

  18. Biosafety Level 2 • Biosafety Level 2 Signs • Policies and procedures for entry • Biohazard warning signs • Specific hazard training with annual updates

  19. Biosafety Level 2Laboratory Facilities (Secondary Barriers) • Hand washing sink located near laboratory exit • Autoclave available • Eyewash/Shower station available • Biosafety Cabinets

  20. Biosafety Level 2 Laboratory Facilities (Secondary Barriers)

  21. Special Practices • Use leak-proof transport containers

  22. Biological Spill Kits • Location • Inside Tissue Culture Rooms • Contents • Gloves, Goggles/shield, N95 Respirator, Lab coat/gown • Absorbent material • Bleach or suitable disinfectant • Tongs and dust pan • Spill Procedures • Place absorbent material over spill • Pour disinfectant around outer edges of absorbent ending in the middle • Wait for inactivation of bio-hazardous material • Properly dispose of all materials in biohazard box

  23. Biohazardous Waste Includes... • Blood and Blood Products • Cultures/Stocks of Infectious Agents and Associated Biologicals • Animal Carcasses and Bedding • Sharps • Glass / Volumetric Pipettes • Primary and Established Human and Animal Cell Lines

  24. BL1 and BL2 Waste DisposalProcedures • Liquid: • chemically treated prior to drain disposal • 10% (total volume) sodium hypochlorite solution (Bleach; Hg free) for 20-30 minutes • Solid: • BL1 and BL2 non recombinant waste • Dispose of directly into red plastic waste containers or cardboard boxes lined with red biohazard bags • Recombinant DNA • Any viable organism containing recombinant DNA must be rendered noninfectious prior to disposal • Steam sterilization or chemical inactivation

  25. BL2S Waste Disposal • Liquid • Inactivate all liquid waste with Vesphene then autoclave • Solid • Autoclave all solid waste • Avoid using Sharps and Glass if possible

  26. Sharps Disposal • All sharps should be disposed of directly into an approved sharps container immediately after use • When the sharps container is 2/3 full: • Place in biohazard waste container • Arrange for pick up from Building Services (x2656)

  27. COMS • Committee on Microbiological Safety • The MGH Institutional Biosafety Committee (IBC) • What research needs COMS approval • Infectious microorganisms, in-vitro or in-vivo • Recombinant DNA materials • Human studies involving rDNA (Human Gene Transfer studies) • Animal to human transplants (xenotransplantation) • Primate tissues • Dual use agents • All COMS approved research is subject to an annual biosafety inspection • The Forms: http://www.hms.harvard.edu/orsp/coms/

  28. OSHA Bloodborne Pathogens Training

  29. Bloodborne Pathogens What are the three bloodborne pathogens you need to be concerned about and what is the risk of transmission each?

  30. BLOODBORNE PATHOGENS • HEPATITIS B (HBV) 30% • HEPATITIS C (HCV) 3 % • HIV 0.3%

  31. Bloodborne Pathogens • Hepatitis B • the most highly transmissible • but greatly reduced by vaccine use!! • Hepatitis C • No vaccine, but early identification and treatment offers some hope • HIV • Early treatment greatly reduces the risk

  32. Exposure Prevention strategies • Vaccination • Work Practices/Standard Precautions • PPE/Engineering Controls • Exposure response • Education/polices

  33. Hepatitis B vaccine • Available from Occupational Health Service (6-2217) • Must be offered to all employees with exposure risk • A 3 shot series- must complete • Antibody status checked after series completed • No need for routine “boosters”

  34. Work Practice Controls • Standard Precautions - Treat all blood and body fluids as infectious! - Gloves for contact with blood, body fluids, moist mucous membranes, non-intact skin, secretions and contaminated items - Other Personal Protective Equipment (PPE) as needed to suit the task you are performing - Remove PPE when task completed and moving to a clean area - Disinfect hands after contact with blood/body fluids/tissue, even if gloves are used - Disinfect equipment/surfaces per policy

  35. PPE/Engineering Controls • Gowns, gloves, face shields, splashguards etc. • Labeling • Biohazardous Waste Disposal • Safety Devices • Sharps Disposal • Sharps Containers

  36. Post Exposure Response • How can you be exposed? -Percutaneous -Mucous membrane -Broken Skin - Animal Bites • Immediate Response -First Aid- wash area • Reporting- Occ. Health (x2438) or call operator (x2000), available 24/7 -All animal bites must be reported to Occ. Health • Follow-up required

  37. Policies and Procedures • Bloodborne Pathogens Exposure Control Plan- located in the Infection Control Manual http://library.partners.org/MGH1/webserver/custom/trov • Departmental Policies • Annual Education

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