Basics of Biosafety
Working Safely with Biological Materials
What is Biosafety?

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
Hierarchy of Controls
The Hierarchy of Controls is the preferred method of controlling preventing and
controlling hazards. You must document that you went through this process.
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Administrative Control: Rotating workers more often
to reduce exposure times
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Engineering Control: Installing guards on machinery,
ventilation, sound proofing
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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
permissible exposure limit
Administrative Controls
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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 history
 Medical screening
 Immunization
 Serum storage
 Post-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)

Biosafety Levels (BSL)
 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 Level 1

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
BSL 1: Work Practices and Procedures


Applications
 Non-infectious agent and tissue culture,
media preparation
Prevent Cross Contamination
 Keep cultures covered
 Flame instruments and containers
 Use sterile media and equipment
 Keep hands or face away from cultures
BSL 1: Work Practices and Procedures

Biosafety Procedures
 Work with agents may be conducted on
open bench
 Wash hands often
 No mouth pipetting
 No eating or drinking in lab
 Minimize aerosol generation
 Decontaminate work surfaces
 Wear applicable PPE
BioSafety Level 2

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
BSL 2: Work Practices and Procedures
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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
BioSafety Level 3

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 4

FORGET ABOUT IT!!!

Hemorrhagic fever, deadly viruses, etc.

Total containment, airtight labs, “submarine”
doors, air pumps, water treatment, etc.

Positive pressure “moonsuits”
Exposure Controls
Careful Pipetting Techniques
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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
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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 Controls
Safe 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 Controls
Personal Protective Equipment (PPE)
• Gloves
- Latex
- Nitrile
- Vinyl
- Utility
Nitrile and vinyl gloves
Exposure Controls
Personal Protective Equipment (PPE)
• Protective clothing
- Lab coat
- Gown
- Apron
- Shoe cover or boot
Exposure Controls
Personal 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
Lettering and symbol
fluorescent orange in contrasting color
or orange/red
to background
background
Biohazardous Waste Categories
 Cultures and stocks of infectious agents
and associated biologicals
 laboratory
waste
 biological production waste
 discarded live and attenuated vaccines
 culture dishes and related materials
 contaminated PPE
Biohazardous Waste Categories

Liquid human and animal waste
 liquid
or semi-liquid blood and blood
products and body fluids
 contaminated
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
 tissues
 body
parts other than teeth
 products 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
 Autoclave
Disposal: 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 containers
that are puncture-resistant,
closable and will capture
leakage, BUT….
…Do NOT use
SHARPS containers!
#3
Is it a sharps hazard?
Examples:
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–
–
–
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 containers
must be used for
both clinic and
field 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 a
one-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 mercury
thermometers
What’s wrong with this picture?
Sharps Container Disposal

Containers must be permanently closed
and disposed:
 Within
90 days
of first use
 When

¾ full
Disposal methods:
 Landfill
‫مكب‬
 Incineration
 We
‫الحرق‬
use waste hauler ‫متعهد للنفايات‬
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
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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