Basics of Biosafety
Working Safely with Biological Materials
Principles and practices employed to protect laboratory personnel and the environment from exposure or infection while working with living organisms, biological materials, or agents.
Included are any materials that may be potentially infectious.
Includes recombinant DNA research
What is Biosafety?
Hierarchy of Controls
Administrative Control: Rotating workers more often to reduce exposure times
Engineering Control: Installing guards on machinery, ventilation, sound proofing
Safe Work Practices: policies and procedures increasing safety either by changing the actual way the work is done or by adding a tool to help
Personal Protective Clothing or Equipment: respirators, ear plugs
The Hierarchy of Controls is the preferred method of controlling preventing and controlling hazards. You must document that you went through this process.
permissible exposure limit
Administrative Controls Authorization/Approval Written biosafety procedures required for the
experimental procedures and equipment including inventory of biological agents or materials
Laboratory personnel biosafety training Medical Surveillance (BSL 2 and above)
Health historyMedical screening ImmunizationSerum storagePost-exposure prophylaxis
Engineering Controls
Biological safety cabinets, glove boxes Animal containment caging systems Safety equipment (filtered or sealed
equipment) Ventilation system Containment facilities
Personal Protective Clothing and Equipment
Provides barrier against skin, mucous membrane or respiratory exposure to infectious agents during procedures
Prevent spread of contamination Integrity wanes with use (i.e., change
gloves frequently)
BSL are used in risk management
BSL are ways to control the agent facilities, safety equipment, practices, PPE, etc.
Once risk is assessed then the appropriate BSL is determined
Biosafety Levels (BSL)
Well characterized, non-pathogenic organisms or agents
Open bench- no containment
Use good laboratory practices, waste disposal, and aseptic techniques (i.e. performed under sterile conditions)
Example: E. coli K-12 strains
BioSafety Level 1
BSL 1: Work Practices and Procedures
ApplicationsNon-infectious agent and tissue culture,
media preparation Prevent Cross Contamination
Keep cultures coveredFlame instruments and containersUse sterile media and equipmentKeep hands or face away from cultures
Biosafety ProceduresWork with agents may be conducted on
open benchWash hands oftenNo mouth pipettingNo eating or drinking in labMinimize aerosol generationDecontaminate work surfacesWear applicable PPE
BSL 1: Work Practices and Procedures
Agents of moderate hazard to personnel or environment
Basic lab, but restricted access, containment during certain processes (i.e. aerosols, large volumes, etc.)
Autoclave and Biological Safety Cabinet desired
Use good laboratory practices, waste disposal, and aseptic techniques
Example: most non-respiratory, non lethal, agents
BioSafety Level 2
BSL 2: Work Practices and Procedures
Increasing emphasis on safety procedures and practices
Increasing need for staff training Increasing need for competent
supervision Biohazard sign posted at entry door Biohazard labels affixed on regulated
waste containers Use of personal protective equipment
as a barrier to exposure: lab coat, gloves, eye and face protection
Some work on open bench allowed
BSL 2: Work Practices and Procedures Aerosol generating procedures
performed in a biosafety cabinet: Homogenizing Vortexing Vigorous mixing Pipetting infectious liquids Sonication Pouring
If breach occurs: Evacuate lab, post spill sign With appropriate PPE and
disinfectant, decontaminate centrifuge, buckets, other items or areas
Agents of high hazard to personnel or environment
Respiratory exotic or indigenous agents which are easily transmissible causing serious or lethal disease
All work is contained, engineering controls and controlled environments
Example: Mycobacterium tuberculosis, SARS, etc.
BioSafety Level 3
FORGET ABOUT IT!!!
Hemorrhagic fever, deadly viruses, etc.
Total containment, airtight labs, “submarine” doors, air pumps, water treatment, etc.
Positive pressure “moonsuits”
BioSafety Level 4
Exposure Controls Careful Pipetting Techniques
Never blow out last drop in pipette Use pipetting aids with filters Never mix by suction and expulsion
(mix by sonication) Discharge liquid down side of
container, using tip-to-wall contact Deliver as close as possible to
contents Work over plastic-backed absorbent
matting (ensure it doesn’t slide forward or backward blocking air grill)
Exposure Controls Use Extreme Care with Sharps
Use sharps if only absolutely required as part of a process
Percutaneous exposure risk Employ safe work practices Utilize safe sharp devices
Aerosol exposure risk Use biosafety cabinet for
removal of air from needle Use mechanical methods for
needle removal Never bend, recap or manipulate
sharps by hand Keep hands away from needle
Exposure Controls
Safe Work Practices Wash hands after
each glove use and immediately or ASAP after exposure.
Remove PPE before leaving work area.
Flush body parts with water after contact with blood or OPIM
OPIM = Other Potentially Infectious Material
Exposure ControlsSafe Work Practices
Clean-up of spills and broken glassware/sharps contaminated with blood or OPIM
• Wear protective eyewear and mask if splashing is anticipated
• Remove glass and other sharps materials using a brush and dust pan, forceps, etc. Do not use your hands
• Properly discard all materials into a sharps or puncture-resistant biohazardous waste container
• Use paper/absorbent towels to soak up the spilled materials
Exposure ControlsPersonal Protective Equipment (PPE)
• Gloves
- Latex
- Nitrile
- Vinyl
- UtilityNitrile and vinyl gloves
Exposure ControlsPersonal Protective Equipment (PPE)
• Protective clothing
- Lab coat- Gown- Apron- Shoe cover or
boot
Exposure ControlsPersonal Protective Equipment (PPE)
• Eye-Face Protection and Masks
- Safety glasses with side shields
- Splash goggles
- Face shield- Mask
Biohazardous/Medical Waste Waste that is potentially infectious to humans, animals or plants
Biohazard Labels and Signs:
• Must have biohazard symbol• Labels attached securely to any
containers Biohazardous /Medical waste
Predominantly fluorescent orange
or orange/red background
Lettering and symbol in contrasting color
to background
Biohazardous Waste Categories
Cultures and stocks of infectious agents and associated biologicals
laboratory wastebiological production wastediscarded live and attenuated vaccinesculture dishes and related materialscontaminated PPE
Biohazardous Waste Categories
Liquid human and animal waste liquid or semi-liquid blood and blood
products and body fluidscontaminated items that would release
blood or items that are caked with blood or other potentially infectious materials; NOT including urine or materials stained with blood or body fluids
infectious animal waste (research)
Biohazardous Waste Categories
Pathological waste tissuesbody parts other than teethproducts of conception fluids removed by trauma or during
surgery or autopsy/necropsy or other medical procedure and not chemically fixed.
…And More Biohazardous Waste Categories
Animal and plant pathogen waste Recombinant DNA waste Sharps
Managing Liquid Biohazardous Waste
Storage: Label and secure bulk
vessels if not disposed of immediately
Treatment: Chemical disinfection OR AutoclaveDisposal: THEN Flush to sewer Use proper PPE!
Managing Non-Sharp Biohazardous Waste
labeled container
lined with a biohazardous waste bag
equipped with a lid.
Managing Non-Sharp Biohazardous Waste
Securely tie bags for transport to treatment/collection site.
When moving wastes, use secondary containment; avoid using public halls and elevators.
“Breakable” Non-sharps Biowaste
Store in labeled containersthat are puncture-resistant, closable and will capture leakage, BUT….
…Do NOT use SHARPS containers!
#3Is it a sharps hazard?
Examples:– needles– syringes– Scalpels مشرط– all biologically contaminated objects that
can easily penetrate skin (Pasteur pipettes,razor blades, etc.)
Place sharps in approved sharps container for disposal!
…Syringes in research settings should be disposed of as a sharp to avoid public relations concerns!
Sharps Containers Containers must be leak-proof,
puncture-resistant, closable & labeled with the biohazard symbol.
Proper sharps containersmust be used forboth clinic andfield work.
Proper Use of Sharps Containers
Place tops on containers before use on lab bench
Don’t forget to date the container when first put into use
Remember: sharps containers are aone-way disposal system
Proper Use of Sharps Containers
Use sharps containers for sharps ONLY!• No solid biohazardous waste (i.e.
gauze, un-broken pipettes, gloves)
• No mercurythermometers
What’s wrong with this picture?
Sharps Container Disposal Containers must be permanently closed
and disposed:Within 90 days
of first useWhen ¾ full
Disposal methods:Landfill مكب Incineration الحرق We use waste hauler للنفايات متعهد
Safety Notes on Sharps Use Do not re-cap sharps Keep sharps container in
close proximity to point of use (i.e. limit handling) for easy disposal
Do not leave needles in pockets of coveralls or smocks
Managing All That Other Waste…
Do NOT discard medications in the trash.
Return to source for disposal or seek assistance.
Recordkeeping
Medical Records• For employees with
exposure• Confidential• Hepatitis B vaccination
status • Post-exposure evaluations• HCP’s written opinions• Information provided to HCP
as required
HCP = Health Care Property Investors
Recordkeeping
Training Records
• Dates• Content summary• Trainer name &
qualifications• Attendee’s names & job
titles