Lab Safety: Everyone Is Responsible!

Safety In the Microbiology Lab

Rules and Symbols

Mr. S. Kashif Raza

Z. A. School of Medical Technology
2006-2011
Al-Quran
 Safety First
• In Microbiology lab many laboratory activities,
require the use of
– Hazardous chemicals
– Infectious microorganisms
– Expensive lab equipment.
• Safety is the #1 priority.
• A list of rules has been developed.
• These rules must be followed at all times.
 Safety First
• Common sense
• Each item in microbiology has potential danger.
• Restrict microorganisms in specimens or cultures
to the vessels.
• Prevent environmental microorganisms
– Hand
– Hair
– Clothing
– laboratory benches
– air
Why does it matter?
• Safe working
protects:
– You
– Other lab workers
– Cleaners
– Visitors
– Your work
– environment
What does the law say? (1)
• Health Safety at Work
etc Act 1974
– You must work safely
– You must not endanger
others
– You must not misuse
safety equipment

• Penalty – up to 2 year
in prison &/or an
unlimited fine
What does the law say? (2)
• The Management of
Health and Safety at
Work Regs 1999
• Control of Substances
Hazardous to Health
Regs 2004
• You must perform
RISK
ASSESSMENTS
 How to do a Risk Assessment?
• Determine
– Hazards
– Evaluate risks
• Use all relevant available data
• Determine controls needed to
minimise risks
• Document the assessment
• Agree it with your supervisor
• Use those control measures
 Routes of infection

• Oral

• Skin

• Conjunctiva

• Lungs
 ORGANISMS
• Risk Category
– All organisms are regarded as potential human
pathogens and treated accordingly.
– Organisms are categorized into
– Risk Group 1
– Risk Group 2
– Risk Group 3
– Risk Group 4
 Accidents in laboratory
• The important diseases/organisms are:
– Hepatitis B,C virus, HIV
– Shigella spp.
– Salmonella spp. including S typhi
– Brucella spp.
– Bacillus anthracis
– Leptospires
– Yersinia pestis
– Mycobacteria spp.
– Histoplasma
 Laboratory biosafety levels
• Bio-safety Level 1 to 4 for Risk Groups 1 to 4
microorganisms respectively.
• Biosafety Level -1 (BSL-1):
– Standard microbiological practices
– No special requirement
• Biosafety Level-2 (BSL-2):
– BSL-1
– Laboratory coats
– Decontamination of infectious wastes
– Limited access
– Protective gloves
– Display of biohazard sign
 Laboratory biosafety levels
• Biosafety Level-3 (BSL-3):
– BSL-2
– Special laboratory clothing
– Controlled access to laboratory
• Biosafety Level-4 (BSL-4):
– BSL-3
– Entrance through change room
– Shower on exit
– All wastes decontaminated
 Laboratory facilities in BSL-2
• Design
– Easily cleaned.
• Sink.
• Laboratory tops
– impervious to water
– resistant to acids, alkalies and organic solvents.
• Autoclave
• Illumination
• Adequate storage space.
 General Safety Guidelines
• Be Responsible at All Times.
• Follow all instructions carefully.
• No horseplay, practical jokes, pranks, etc.
• Do not play with lab equipment until
instructed to do so.
• Food, drink, smoke and gum are not
allowed in the microbiology lab .

Lab Safety: Everyone Is Responsible!

 General Safety Guidelines
• Don’t lick labels.
• Don’t drink from laboratory glassware
• Don’t wander about the laboratory; cause:
– Accidents
– Distract others
– Promote contamination
• Operate centrifuges, homogenizer and shakers
safely
• Immunize the laboratory workers
 General Safety Guidelines
• Keep the microbiology lab clean
and organized.
• Notify supervisor immediately
of any accidents or unsafe
conditions in the microbiology
lab !
• Wash your hands with soap and
water after work.

Lab Safety: Everyone Is Responsible!

 General Safety Guidelines
• Listen to or read instructions
carefully before attempting
anything.
• Wear safety goggles to protect
your eyes from chemicals,
heated materials, or things that
might be able to shatter.
• Notify your teacher if any spills
occur.
 General Safety Guidelines
• Keep your hands away from your face.
• Tie back long hair away from shoulders .
• Wear glasses rather than contact lenses.
• Never put into your mouth
– Pencils
– Pens
– fingers etc
• Clean up your lab area at the conclusion
of the laboratory period.
 General Safety Guidelines
• Roll up loose sleeves.
• Button-up your Lab Coat
• Lab coat thickening.
• Know the location of the
fire extinguisher, Gas
isolation switch, fire
blanket, eyewash station,
first aid kit & Exits.
• Keep your work area
uncluttered.
 General Safety Guidelines
• Take to the lab station what is
necessary, Left outside
– Coats
– Jackets
– Bags and books not required
for the laboratory session.
• Cover any open cuts on hands
and other exposed skin surfaces
 General Safety Guidelines
• Wear mask N95 4 mycobacterium.
• Do not wear any jewellery.
• Carry out procedures to minimize risks of :
– Spills
– Splashes
– Production of aerosols.
• No slides or cultures are to be taken from,
or brought into the lab without permission.
 General Safety Guidelines
• Don’t use your Mobile.
– Medical devices
– Chemicals
– With gloves
– RF interference
• Do not attend calls with gloves.
• Writing
– Without gloves
– Working area
 General Safety Guidelines
• Avoid wearing artificial nails
• Keep natural nails <1/4 inch
• Foot wear that completely
covers the foot is required
• No Open shoes
• No rings or hand accessories
• Do not wear loose clothing that
could catch on fire
• Do not open doors with glows
 ACCESS
• Understanding of the safety practices.
• The laboratory doors, closed at all times
• Outer door must be locked unless a staff member
is on the premises.
• Students not permitted to work without a staff
member and direct supervision when authorized
by a Microbiology Safety Officer.
– Written form
– Sighted by the staff member designated to be
present in the laboratory.
•  
 ACCESS
• No visitors into the laboratories without
permission.
• Only authorized visitors shall enter the laboratory
showing universal biohazard sign
• Appropriate signs should be located at points of
access to laboratory areas directing to
– Receptionist or receiving officer
 ACCESS
• The universal biohazard symbol displayed
at specific laboratories in which
manipulations of organisms with moderate
and heavy risk.
• Doors displaying biohazard symbol shall
not be propped open, but shall remain
closed when in use.
 PERSONAL PROTECTIVE
EQUIPMENT and PROCEDURES
• Gowns
• Safety Glasses and Eye Protection
• Gloves
• Mask
• Hand washing
• High Risk Individuals /Antenatal
Considerations
 Gowns\Coats
• Rear opening wrap around gowns.
• Laundered every fortnight during semester.
• Soiled gowns in excess of "normal use" are to
be removed from circulation.
• Students wear gowns at all times in the
laboratory.
• Laboratory gowns must be worn inside the
laboratory but neat & clean.
 Gowns\Coats
• Not to be worn outside the
laboratory for any reason.
• At the end of the laboratory
session
– Return the gown to the
hook
– Neatly folded
– Inside out
 Gloves
• For routine work, not essential.
• Worn under the following circumstances;
– Mopping up a spill.
– Where there is a high risk of contaminating
hands.
– If open cuts or skin conditions.
– When instructed by the demonstrator
– Adequate supplies of gloves. If you use the
last gloves, please notify so that stocks can be
replenished.
 Safety Glasses and Eye Protection
• “Safety glasses" for all practical sessions,
– Risk of splashing with infectious or corrosive
liquids.
– When opening the autoclave.
• An eyewash station.
– “Fresh" sterile distilled water
– Replace every month
– Record date of "renewal"
 Hand washing
• The standard hand washing procedure is to
use running water and "Hibiclens".
– Must before leaving the laboratory.
• The "Hibiclens" dispensers at all hand
washing sinks within the laboratory must
always be sufficiently full.
Hand washing
 High Risk Individuals /Antenatal
Considerations
• Immunocompromised or
particularly susceptible to
infection need to take additional
precautions.
– Pregnant women, so any
female student or staff
member should discuss the
matter with the academic in
charge.
• Student with medical condition. 
• Children.
 Chemical Safety
• Do not inhale the fumes.
• Never pour water into a
concentrated acid.
• Wash hands.
• Wear goggles
• Lab apron
• Never taste any chemicals
(never taste anything).
 Electrical Safety
• Lay electrical cords where no one can
trip on them or get caught in them.
• hands dry before using electrical
equipment.
• Never poke anything into electrical
outlets.
• Unplug cords by pulling the plug and
not the cord.
• Unplug all electrical equipment at the
end of the lab period.
 Heating Safety
• Let burners and hotplates cool down before
touching.
• Use tongs and/or protective gloves.
• Never reach across an open flame.
• Wait until the striker is in place before you
turn on the gas.
• Heating a test tube, move it around slowly.
• Dry glassware should be heated.
• Do not hold it in your hand
• Never leave a burner or hotplate unattended.
LABORATORY EQUIPMENT
• Autoclave
• Biohazard Cabinet
• Gas Supply
 Autoclave
• To avoid Noxious fumes turn on the extraction
fan.
• Safety glasses
• De-pressurized before opening.
• Heat safety gloves.
• Students under the direct supervision.
• Students must be given instruction before using.
 Biohazard Cabinet
• Class II cabinet to protect the operator.
– Close doors to minimize the disruption of the
airflow.
– allow it to run motor for 30 sec.
– Wipe down the interior work surfaces.
– Either install the shield or leave the room if
the UV lamp is required.
– Check annually by Laminar Airflow Services.
– Notify about any major spills.
 Pipetting
• The significant and consistent cause of
occupational infections.
– Reduce creating aerosols.
– Plug pipettes with cotton.
– Avoid rapid mixing of liquids by alternate
suction and expulsion.
– Don’t forcibly expel material from a pipette.
– Don’t bubble air through liquids.
– Prefer pipettes that do not require expulsion of
last drop of liquid.
 Pipetting
• Drop material close to the fluid or agar
level.
• Place contaminated pipettes in a container
having suitable disinfectant immersed.
• Selection depend upon ease of operation
and type of work.
• Do not place contaminated pipettes on the
bench top.
 Gas Supply
• Each room has an isolation switch.
• Incase of a spill of flammable liquid, fire or back
burning of an individual Bunsen burner, de-
activate the isolation switch immediately.
• A master isolation switch to the entire laboratory,
should be de-activated at the end of each day.
• The gas supply is to remain off until a staff
member declares it safe to switch it back on.
 Transport
• When viable organisms
– Cultures
– Specimens
– stock organisms are transported to or from the
laboratory, the "primary" container 
"secondary" container.
• The secondary container sealable and non
breakable.
 Storage and Identification
• Cultures should be suitably identified
– degree of risk can be ascertained.
• For "unknown" cultures at least two people
shall be aware of the true identity.
– Practical demonstrator
– Laboratory Technician
  Hypodermic syringes and needles
• Common causes of occupational infections.
– Collection
– Processing
– 25%.
• Avoid quick & unnecessary hand movements.
• Examine for chips and cracks, and needles for barbs
and plugs.
• Use needle locking syringes.
• Wear gloves.
• Fill syringes carefully to minimize air bubbles and
frothing.
 Hypodermic syringes and needles
• Expel air, liquid & bubbles vertically into a
disinfected cotton swab.
• Don’t forcefully expel into an open vial for mixing.
• Mixing if the tip is held below the surface of the fluid.
• Don’t manipulate by hand.
• Place directly into a puncture-resistant container.
• Decontaminate before
– Disassembly
– Reuse
– Disposal.
 Opening containers          
• Opening of vials, flasks, Petri dishes, culture tubes
& other containers poses potential risks of creating
– Droplets
– Aerosols
– Contamination of the skin or the immediate
work area
• The most common is the removal of stoppers from
containers of clinical materials.
• Received & opened only by personnel with
knowledge risks.
 Opening containers
• Well-lighted and designated areas.
• Wear coat & gloves.
• absorbent paper towel to:
– facilitate clean-up
– reduce generation of aerosols
• Leaking or broken Specimens in BSC.
• Tubes containing bacterial cultures, handle with care.
• Vigorous shaking of liquids & when sealed ampoule
containing a lyophilized or liquid culture is opened,
aerosols may be created
CLEANING & Waste Disposal
• Separate the non-infectious from the
infectious waste.
• Dispose the infectious waste to
– Minimizes the risk to both staff and students
– Facilitates the recycling of reusable material.
 Sharps
• “Yellow" sharps container at the back of each
laboratory.
– Needles
– scalpel blades
– other sharp materials
• Always move the "sharps container" to your
work place to dispose of such items.
• Do not wander around the laboratory carrying
sharps.
 Biogram Buckets
• Containers of general purpose phenolic,
hypochlorite (10 gm/L) laboratory disinfectant
• Used for disposal of small contaminated items
– used swabs
– capillary tubes
– wet slides
– Pipettes
– inoculated reagent strips
– glass culture tubes.
 Biogram Buckets
• Disposing pipettes, place tip first into the
biogram
• Not for Gram stains, non contaminated paper or
matches.
• 1/40 dilution of the commercial product 18%
phenol equivalent.
• Change weekly; or if grossly soiled.
 Biogram Buckets
• autoclaved prior to removing or disposing of the
contents.
• Allow the buckets to cool before emptying the
contents (sieve) down the drain.
– Solid materials collected can then be bagged
and discarded.
– Glass and other sharp objects should be
disposed of in the bin for "broken glass".
 Biohazard Bin
• Centre of each laboratory.
• For the disposal of contaminated waste, e.g.
– Used culture plates
– Contaminated paper towel.
• Not to be used sharps & Any non contaminated
paper wastes.
• Do NOT use these bins for
– Paper towel discarded after hand washing
– Blotting paper discarded after blotting slides.
• autoclaved before disposal.
• Left open to allow penetration of the steam
 Billy Cans
• Two stainless steel tins at the front of each
laboratory.
• Container 1:
– Contaminated and non contaminated
recyclable glass or plastic tubes.
– Before placing items ensure that they are
capped and that sticky labels are removed.
• Container 2:
– Fixed and stained slides
– Not to be used for "wet slides" & emdash;
 Paper Bin
• To be used for non contaminated paper
waste only, e.g..
– Paper towel from hand washing (but not bench
wiping)
– Blotting paper from blotting Gram stains.
 Broken Glass
• When any glassware is broken, notify the
instructor immediately for assistance with
disposal.
– Non contaminated can be disposed of in the
Billy cans
– Contaminated should be placed into a stainless
steel Billy and autoclaved prior to disposal in
the glass bin.
 Sink
• Don’t place any hazardous or infectious
materials.
• Don’t dispose of any solid material.
 CLEAN-UP PROCEDURE FOR
BIOHAZARD SPILLS
• Spills involve infectious materials, are complex.
• No set of instructions
• sound microbiological judgment in their
management.
• All must be familiar with the guidelines.
• The order of priorities is as follows:
– Protection of personnel
– Confinement of contamination
– Decontamination of personnel
– Decontamination of area involved
 General considerations for
biohazard spills
• Dispersed into three spill fractions:
– Puddle.
– Splashes.
– airborne particles (aerosols)
• airborne particles pose the greatest risk, remain
airborne and be dispersed to other areas.
• General purpose laboratory disinfectant
(Biogram) can be used
 Diversol

• Spills involving human blood or body fluids,

• 5000 ppm of available chlorine.
• Limited shelf life.
• Sachets stored on the chemicals shelf in the
preparation room.
• 8.3 Squirt Bottles
• Sufficient "biogram squirt bottles".
• Clearly labelled.
 Assessing a Biohazard Spill
• The designation into
– "minor"
– "major"
• The supervising staff member responsible.
– The biological nature.
– The physical nature
– The volume. 
 Minor biohazard spills
• Minimally hazardous material.
• Low potential for generation of aerosols.
• If hands contaminated, first wash hands.
• Remove and replace any contaminated protective
clothing.
• Put on gloves.
• Lay down absorbent material wetted with
disinfectant over the spill and allow to sit for 10
minutes.
•  
 Minor biohazard spills
• Discontinue working
• After 10 minutes, mop up spill and place
contaminated materials into autoclave bag.
• Wipe over general area again with paper towel
dampened with disinfectant.
• Remove gloves and wash hands.
 Major biohazard spills
• Major risk with larger volume
• Considerable production of splashes and aerosols.
• Hold breath
• Warn others
• Leave the room immediately.
• Close doors and place a "DO NOT ENTER" sign
on the door.
• Remove any contaminated clothing and wash any
contaminated body surfaces.
• Notify MSO (preferably) or other senior.
 Major biohazard spills
• "Spill Clean Up Team"
– two to clean up
– one to supervise.
• "gowns, gloves, face masks and safety glasses" before
entering the spill area.
• Re-enter after 30 min.
• Do not pour disinfectant directly onto the spill
• Lay paper towels wetted with disinfectant onto the
spill for 30 min.
• Disinfectant areas around the spill, likely to have been
contaminated.
REPORTING OF INCIDENTS

• All accidents and major spills should be

documented on an "incident report form" through
the relevant Departmental Occupational Health
and Safety Officer
• Copies of any documentation relating to an
incident or safety hazard must be sent to both
MSO and a copy should also be retained.
 First Aid
Injury: Burns
What To Do: Immediately flush with
cold water until burning
sensation is lessened.
 First Aid
Injury: Cuts, bruises
What To Do:
• Do not touch without
gloves.
• Pressing directly on minor
cuts.
• Apply cold compress to
bruises to reduce swelling.
 First Aid
Injury: Fainting
To Do:
Provide fresh air and have
the person recline so that
their head is lower than the
rest of their body.
 First Aid
Injury: Eyes
What To Do:
• Flush eyes immediately
with plenty of water for
several minutes.
• If a foreign object is
lodged in the eye, do not
allow the eye to be rubbed.
 First Aid
Injury: Poisoning
What To Do: Find out what substance
was responsible for the
poisoning and alert the
teacher immediately.
 First Aid
Injury: Spills on the skin
What To Do: Flush with large
quantities of water. For
acid spills, apply baking
soda solution. For base
spills, apply vinegar or
boric acid.
 First Aid
Injury: Electrical shock
What To Do:
• Shut off the current at the
source.
• Remove wire with rubber
gloves.
• Alert the teacher immediately.
What’s Wrong With This
Picture?
What’s Wrong With This
Picture?
What’s Wrong With This
Picture?
What’s Wrong With This Picture?
 When in doubt – ASK!!!
• Do not carry out a
new or unfamiliar
procedure until you
have been fully
trained & understand
the precautions
necessary for safe
working
• DO NOT GUESS!!!!
 Reference
• Food Standards Agency - Microbiological safety. html
• Guidelines on Standard Operating Procedures for MICROBIOLOGY -
Safety in Laboratories. tm WHO
• Microbiology Rules for Using Disinfectants eHow com. htmL
• Microbiology Safety and Staff Induction Manual University of Tasmania -
Launceston Campus Version 2, February 2000
• Mrs. Page Kennedy Middle School 2002-2003
• Presented by: Leonard LaFazia