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

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Presentation on theme: "1 Biosafety in Research Laboratories. Definitions Biohazard An agent of biological origin that has the capacity to produce negative effects on humans,"— Presentation transcript:

1 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 ) l Perform experiments in biological safety cabinets l Use respiratory protection Remaining 20% LAI are a combination of: §Percutaneous l Avoid sharps when possible, use plastic pipettes and disposable glassware §Mucous Membrane (eyes, nose and mouth) l Wash hands often, avoid touching face §Ingestion l 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 l Use attenuated or non-virulent strains, require restraint during animal injection, avoid sharps and glassware, require decontamination, Standard Operating Procedures, training §Engineering Controls l Biosafety Cabinets, Sealed Rotors/Safety Cups, Pipette Aids, Safety Enclosures for Cell Sorters §Personal Protective Equipment l Lab Coat / gown, gloves, goggles, facial protection l 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 lProtection against particulates only lNo gas or vapor protection lAir flow is easily disrupted Dont cover the grill Move arms slowly in and out of cabinet Walk slowly l70% 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 l Minimize splashes and aerosols l PPE l Decontaminate work surfaces daily l Decontaminate wastes l 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 1 Laboratory 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 l Biosafety Level 2 Signs Policies and procedures for entry Biohazard warning signs l Specific hazard training with annual updates

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



25 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

26 BL1 and BL2 Waste Disposal Procedures §Liquid: l chemically treated prior to drain disposal l 10% (total volume) sodium hypochlorite solution (Bleach; Hg free) for 20-30 minutes §Solid: l 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

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

28 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: lPlace in biohazard waste container lArrange for pick up from Building Services (x2656)

29 COMS Committee on Microbiological Safety The MGH Institutional Biosafety Committee (IBC) What research needs COMS approval l Infectious microorganisms, in-vitro or in-vivo l Recombinant DNA materials l Human studies involving rDNA (Human Gene Transfer studies) l Animal to human transplants (xenotransplantation) l Primate tissues l Dual use agents §All COMS approved research is subject to an annual biosafety inspection §The Forms:

30 OSHA Bloodborne Pathogens Training

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


33 Bloodborne Pathogens §Hepatitis B lthe most highly transmissible lbut greatly reduced by vaccine use!! §Hepatitis C lNo vaccine, but early identification and treatment offers some hope §HIV lEarly treatment greatly reduces the risk

34 Exposure Prevention strategies §Vaccination §Work Practices/Standard Precautions §PPE/Engineering Controls §Exposure response §Education/polices

35 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

36 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

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

38 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

39 Policies and Procedures §Bloodborne Pathogens Exposure Control Plan- located in the Infection Control Manual ov §Departmental Policies §Annual Education

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