Biosafety Training - McMaster Faculty of Health Sciences

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Transcript Biosafety Training - McMaster Faculty of Health Sciences

BSL 1 Biosafety
FHSc. Safety Office
Health Sciences Center 1J11, ext. 24956
Print out the accompanying test and return
(in handwriting) to 1J11 HSC or
fax to 905 528 8539
How to find us
HSC 1J11
Information for undergrad courses
• You will be contacted only if you have
failed your quiz, <75%. Your instructional
assistant will be contacted at the same
• You will not be permitted to attend the
biological labs of your course until you
Biosafety in Canada: Standards
Public Health Agency of Canada (PHAC)
through the “Laboratory Biosafety
Canadian Food Inspection Agency (CFIA)
has “Veterinary Standards for Animal
Human Pathogens and Toxins Act C-11
Ministry of Environment Canada (MOE) for
waste disposal
PHAC has “Laboratory Biosafety
• Sets the standards for work with biological
agents such as bacteria, viruses, fungi
parasites, prions and hazardous cell lines.
• The health risk associated with an
organism will determine the protocols
which must be followed to work with it.
Level of Hazard: BSL 1,2,3,4
Internationally and in Canada, there are four
levels of biosafety, each associated with a
type of lab and specific equipment.
The personal protection required increases
with the level of hazard.
Labs, techniques and methods required
increased safety with the level of hazard.
Risk evaluation to determine
level of use
Risk evaluation of an agent is part of the
responsibility of the supervisor for the
project and is reviewed by the University
Presidential Biosafety Advisory Committee.
All individuals working with an agent must
comply with the level of risk.
Factors affecting risk
• agent in use eg. specific bacterium
• pathogenicity eg. degree of disease
• infectious dose eg. # organisms to cause
• mode of transmission eg. directly by blood
or indirectly through the air
Factors affecting risk
• techniques in use
• host range eg. does it infect animals as
well as humans
• effective treatment or prevention eg. is
there a vaccine?
• Potential for aerosol creation eg.
sonication, vortexing or homogenization
Factors affecting risk
Volume of material eg > 10 liters
• Concentration of material
• Stability in the environment eg. will it
survive on a benchtop?
• Use of re-combinant DNA eg.
oncogenicity, replication ability, host
range, ability to revert to wild type
Biosafety Level 1 (BSL 1)
Low individual
Low community
• unlikely to cause disease in healthy
workers or animals
• treatment / prevention is available
Biosafety Level 2 (BSL 2)
community risk
• causes human or animal disease
• under appropriate use, is unlikely to cause death
to healthy laboratory workers or animals
• treatment / prevention is available
Biosafety Level 3 (BSL 3)
community risk
• usually causes serious human or animal disease
including death. Usually spreads by direct
contact of blood or body fluids from one
individual to another
• generally, no cure / prevention available
Biosafety Level 4 (BSL 4)
• human disease including death and may be
readily transmitted through casual contact
• no treatment / prevention available
Which level of organisms are
being used in your lab ?
• Ask to see the biological equivalent of an
“MSDS” for the agents you will be using.
• Or look up the information on the Internet
site for the supplier of the product.
• Or reference a large database such as
Basic Safety Practices - BSL 1
These requirements must be followed in all
labs working with biological agents.
Know and understand them.
Basic Safety Practices
Longer hair is to be restrained so that it
cannot come into contact with hands,
specimens, containers or equipment.
Oral pipetting of any substance is prohibited
in any lab, use only mechanical fluid transfer
Basic Safety Practices
No eating or storage of food or drink in
the labs…dispose of any food wrapping or
containers BEFORE you enter the lab,
including water bottles and gum.
Keep personal items out of the working area
of the lab.
Basic Safety Practices
It is wise to report any increased
susceptibility to infection to your supervisor
or medical advisor.
Ask the lab supervisor if you are unclear
about any hazard or process.
Dispose of all biological waste in the
designated container only.
Waste disposal
• Biological waste is to be disposed of into
the yellow or red bags in the biohazard
labeled cardboard boxes
• Sharps (scalpel blades, razor blades,
needles on syringes) must be disposed
into a sharps container and then disposed
as biological waste when ¾ full
Basic Safety Practices
Open wounds, cuts, scratches and grazes
should be covered with waterproof
dressings BEFORE working with any
biological material.
Limit use of sharps and glass.
Basic Safety Practices
Minimize aerosol creation
Label all biological material
Where possible and appropriate, protect
yourself by immunization esp Hep B when
working with blood or body fluids.
Personal protection for BSL 1
Protective laboratory clothing must be
worn in the lab but not worn out of the lab
- closed toed footwear
- wear lab coats at all times
- goggles – if using contact lenses
- if required by the protocol
Personal protection for BSL 1
Gloves…latex for water based materials
…nitrile or vinyl for organics
Change gloves when contaminated.
Wash your hands frequently, particularly
when leaving the laboratory.
Staff must understand all hazards with which
they work, including emergency response.
read and know the emergency and spill
response protocol posted in your lab
report any spills or injuries to your supervisor
know the location of spill kits and first aid kits
know the location of eyewash and shower
personnel must receive training on
biohazards and safe handling protocols
- pass for biotraining is 75%
- local lab training to be signed by trainee
and trainer
- retraining program includes annual update
Physical lab requirements for
Physical requirements are a functional and
well designed lab
Doors to the laboratory must not be left
Working requirements for BSL 1
• Basic microbiology lab practices, keep the
lab organized and clean
• Open bench is suitable
• Routine decontamination techniques to
maintain sterility - contamination of
cultures is a risk in an unorganized lab
Supervisor responsibilities for
Access to the laboratory is restricted to
authorized personnel
ie. those who have been advised of the
potential hazards, trained and approved
by the supervisor
Supervisor responsibilities for
Post the lab with a biohazard sign
indicating the biosafety Level and
organism(s) in use
Contaminated materials or equipment
leaving the lab must be appropriately
decontaminated first
Supervisor responsibilities for
All materials must be contained so that they
will not be released to the environment.
Disinfectants effective against the material in
use, must be available at all times.
Have available leak proof containers for
transporting materials between labs.
Supervisor responsibilities for
Maintain an effective insect and rodent
control program.
Investigate spills and accidents, keep
records, use results in future training.
Supervisor responsibilities for
Biological safety information, such as a
manual, must be available
- with procedures for the lab
- it must be read and followed by all
- it must be updated regularly
Supervisor responsibilities for
Provide MSDSs or equivalent.
Ensure compliance within lab.
Any use of animals must comply with the
CCAC and any training requirements of
the CAF and follow the SOP.
Additional biosafety lab
1) Bunsen burners
2) Ultraviolet light
3) Causes of contamination
4) airflow diagram of a biological cabinet
5) airflow diagram of a fume hood
6) airflow diagram of a clean air bench
Bunsen Burners
• Do not use combustibles or flammables in a
biological cabinet – 70% recirculated will allow
buildup of leaking gases
• Upward hot air flow disrupts downward laminar
• May create holes in the HEPA filters
• Splashes may occur (when using loops) and
could cause contamination
• Micro incinerator
• Alcohol burner
Ultra Violet Light
UV lights are not recommended:
• they provide a false sense of security - not giving
off UV light (just prior to the end life of the bulb)
• only disinfects the surface – could contact a
layer of media protein covering the pathogen
• casts a shadow, leaving areas that remain
unexposed to the UV wavelengths
• could cause burns / damage eyes – turn off
when working in the room
Causes of Contamination
1) YOU!
hands not clean, bacteria/spore under your nails
gloves – not sterile from the box
dirty lab coats
lab coat sleeves should be taped
place all materials (cleaned) required into hood
2) Incubator
Avoid dip in line from CO2 tank
Clean humidity source regularly
Clean all surfaces after spill, including holes on trays
If at ground level – tape bench paper
onto floor
Class II Biological Safety Cabinet
use with BSL 2 agents
yes - personnel protection
yes - product protection
yes - environmental
Chemical Fume Hood
do not use with biological agents
yes - personnel protection
no - product protection
no - environmental
Clean Air Bench
use only with BSL 1
no - personnel protection
yes - product protection
no - environmental
Information for McMaster
• Do not ship any samples, or take samples
with you on a trip. Check with Mike
McGuire to ensure it does not require
Transportation of Dangerous Goods
• File a Materials Transfer Agreement with
Bertha Monrose before you share material
with another institution
Information for McMaster
• Do not transform your own body cells.
The immune protection you would
normally have has now been bypassed
• Obtain a biosafety approval number from
the Biosafety office, HSC 1J11 ext 24956
before importing material or working with
biological material
Information for McMaster
• Cell lines can be BSL 1 or BSL 2. Check
with your supplier or your supervisor.
• If you are isolating primary cell lines from
ill humans or animals, they may be BSL 2
until they are well characterized. Check
with the Biosafety office first.
Use of a biological cabinet
• Some labs may chose to use Class II
biological cabinets to maintain sterility for
their BSL 1 agents.
• The following slides cover information for
the use of biological cabinets.
Before you begin
• Check to ensure you are using the right
cabinet for your work.
• Check to see it has been certified in the
last 12 months.
• Ensure the UV light is off anytime anyone
is in the room.
• To start, let the cabinet run for 3 minutes
to purge air.
Before you begin
• Use a disinfectant that will kill any
organism in the cabinet
• Check to see that the window is in the
correct position
• Do not place pens or paper inside the
• Never block the grilles
• Segregate clean items from dirty ones
Before you begin
• Wear a lab coat and tape the coat at the
• Pull the gloves over the taped coat
• Sterilize the gloves with a 70% alcohol
• As you place items into the hood, spray
them down with alcohol to sterilize
Before you begin
• Place sterile items on the left side
• Place items from the incubator at center
• You may place a work pad soaked in
disinfectant on the center front work area
• On the right side, place a waste bucket for
fluids, one for sharps, one for solids
Working in the cabinet
• Keep lab doors closed and keep others
out of the room
• Only one person working in the cabinet
• Use good aseptic technique
• Slow movements directly in or out
• Keep sterile and non-sterile items
• Do not use Bunsen burners in cabinet
Working in the cabinet
• Have posted spill protocol nearby
• No gum, food or drink
• See the two powerpoint presentations on
Good Culture Practices and
Misidentification and Contamination, to
learn more about good practices and why
they are important.
When you are finished
• Spray each item with a disinfectant as you
take it out of the cabinet
• Fill the liquid waste to the max fill line to
achieve appropriate concentration. Mark
on the time when it can be disposed
• Seal the top of the solid waste bag, spray
it, put it into the biohazard cardboard box
• Close the top of the sharps container,
leave it in the hood
When you are finished
• Spray all interior surfaces with the
disinfectant, remember to spray the plastic
window facing you.
• If possible, leave the cabinet running.
• If not, let it purge for 3-4 minutes.
• If there is anything unusual, tell your
supervisor and leave a note for the next
person who will be using the hood.
Suggested layout for working in a Class II biohood
new materials
to go back into
from incubator
F wet material
solution (bleach)
Clean ----------------------- Dirty
Contact information
For additional training videos, biosafety
approval, import permits, reporting of spills
or lab acquired infections, or general
FHSc. Safety Office
Health Sciences Center 1J11, ext. 24956
Carol Carte or Karin Cassidy