1 biosafety in research laboratories. definitions biohazard an agent of biological origin that has...
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Biosafety in Research Laboratories
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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
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What is Biohazardous Material?
BacteriaVirusesParasitesFungiHuman and Non-Human Primate MaterialRecombinant DNAAnimalsBiological Toxins
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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
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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 SortersPersonal Protective Equipment
Lab Coat / gown, gloves, goggles, facial protection Close toed shoes
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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)
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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
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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
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Biological Safety CabinetsMust 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
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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
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Centrifuges
Safety Cups and RotorsLoad and unload safety cups inside the BSC
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Vacuum Line HEPA Filter Protection
Placed in between the vacuum system and the aspiration flasks
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Biosafety LevelsBSL-1: agents not known to cause disease (in
healthy adults)BSL-2: agents associated with diseaseBSL-3: indigenous/exotic agents
associated with human disease and with potential for aerosol
transmissionBSL-4: dangerous/exotic agents of life
threatening nature
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Biosafety LevelsBSL-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)
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Biosafety Level 1Immunocompromised 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)
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Biosafety Level 1 Laboratory Facilities (Secondary
Barriers)
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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
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Biosafety Level 2
Biosafety Level 2 Signs• Policies and
procedures for entry• Biohazard warning
signs Specific hazard training with
annual updates
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Biosafety Level 2Laboratory Facilities (Secondary Barriers)
Hand washing sink located near laboratory exit
Autoclave available Eyewash/Shower station
available Biosafety Cabinets
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Biosafety Level 2 Laboratory Facilities (Secondary Barriers)
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Special Practices
Use leak-proof transport containers
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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
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Biohazardous Waste Includes...
Blood and Blood ProductsCultures/Stocks of Infectious Agents and
Associated BiologicalsAnimal Carcasses and BeddingSharpsGlass / Volumetric PipettesPrimary and Established Human and Animal
Cell Lines
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BL1 and BL2 Waste Disposal Procedures
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
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BL2S Waste Disposal
• Liquid• Inactivate all liquid waste with Vesphene then
autoclave
• Solid• Autoclave all solid waste
Avoid using Sharps and Glass if possible
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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)
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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/
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OSHA Bloodborne Pathogens Training
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Bloodborne Pathogens
What are the three bloodborne
pathogens you need to be
concerned about and what is the
risk of transmission each?
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BLOODBORNE PATHOGENS
HEPATITIS B (HBV) 30%
HEPATITIS C (HCV) 3 %
HIV 0.3%
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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
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Exposure Prevention strategies
VaccinationWork Practices/Standard PrecautionsPPE/Engineering ControlsExposure responseEducation/polices
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Hepatitis B vaccineAvailable from Occupational Health Service
(6-2217)Must be offered to all employees with
exposure riskA 3 shot series- must completeAntibody status checked after series
completedNo need for routine “boosters”
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Work Practice ControlsStandard 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
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PPE/Engineering Controls
Gowns, gloves, face shields, splashguards etc.
LabelingBiohazardous Waste DisposalSafety DevicesSharps Disposal
Sharps Containers
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Post Exposure ResponseHow can you be exposed? -Percutaneous -Mucous membrane -Broken Skin
- Animal Bites Immediate Response -First Aid- wash areaReporting- Occ. Health (x2438) or call operator
(x2000), available 24/7-All animal bites must be reported to Occ. Health
Follow-up required
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Policies and Procedures
Bloodborne Pathogens Exposure Control Plan- located in the Infection Control Manual
http://library.partners.org/MGH1/webserver/custom/trov
Departmental PoliciesAnnual Education