Microbiology Laboratory Safety Rules

Safety In the Microbiology LabRules and Symbols

Lab Safety: Everyone Is Responsible!

Mr. S. Kashif RazaZ. 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 Health Safety at Work etc Act 1974etc 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 RISK ASSESSMENTSASSESSMENTS

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.

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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 AidInjury: EyesWhat 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!!!!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