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


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 .

C virus.Accidents in laboratory ‡ The important diseases/organisms are: ± Hepatitis B. HIV ± Shigella spp. including S typhi ± Brucella spp. ± Bacillus anthracis ± Leptospires ± Yersinia pestis ± Mycobacteria spp. ± Salmonella spp. ± Histoplasma .

‡ 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 ‡ Bio-safety Level 1 to 4 for Risk Groups 1 to 4 microorganisms respectively.

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 .

‡ Autoclave ‡ Illumination ‡ Adequate storage space. . ‡ Sink. alkalies and organic solvents. ‡ Laboratory tops ± impervious to water ± resistant to acids.Laboratory facilities in BSL-2 ‡ Design ± Easily cleaned.

smoke and gum are not allowed in the microbiology lab . ‡ Follow all instructions carefully. ‡ Do not play with lab equipment until instructed to do so.General Safety Guidelines ‡ Be Responsible at All Times. practical jokes. Lab Safety: Everyone Is Responsible! . drink. ‡ No horseplay. etc. ‡ Food. pranks.

cause: ± Accidents ± Distract others ± Promote contamination ‡ Operate centrifuges. homogenizer and shakers safely ‡ Immunize the laboratory workers . ‡ Don¶t drink from laboratory glassware ‡ Don¶t wander about the laboratory.General Safety Guidelines ‡ Don¶t lick labels.

Lab Safety: Everyone Is Responsible! . ‡ Notify supervisor immediately of any accidents or unsafe conditions in the microbiology lab ! ‡ Wash your hands with soap and water after work.General Safety Guidelines ‡ Keep the microbiology lab clean and organized.

‡ Wear safety goggles to protect your eyes from chemicals. or things that might be able to shatter. ‡ Notify your teacher if any spills occur. heated materials.General Safety Guidelines ‡ Listen to or read instructions carefully before attempting anything. .

Never put into your mouth ± Pencils ± Pens ± fingers etc ‡ Clean up your lab area at the conclusion of the laboratory period.General Safety Guidelines ‡ ‡ ‡ ‡ Keep your hands away from your face. Wear glasses rather than contact lenses. . Tie back long hair away from shoulders .

General Safety Guidelines Roll up loose sleeves. ‡ Keep your work area uncluttered. eyewash station. fire blanket. first aid kit & Exits. Know the location of the fire extinguisher. Button-up your Lab Coat Lab coat thickening. ‡ ‡ ‡ ‡ . Gas isolation switch.

General Safety Guidelines ‡ Take to the lab station what is necessary. ‡ Cover any open cuts on hands and other exposed skin surfaces . Left outside ± Coats ± Jackets ± Bags and books not required for the laboratory session.

General Safety Guidelines ‡ Wear mask N95 4 mycobacterium. ‡ No slides or cultures are to be taken from. ‡ Do not wear any jewellery. ‡ Carry out procedures to minimize risks of : ± Spills ± Splashes ± Production of aerosols. or brought into the lab without permission. .

± Medical devices ± Chemicals ± With gloves ± RF interference ‡ Do not attend calls with gloves.General Safety Guidelines ‡ Don¶t use your Mobile. ‡ 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 .

‡ . ‡ Students not permitted to work without a staff member and direct supervision when authorized by a Microbiology Safety Officer. ‡ The laboratory doors. closed at all times ‡ Outer door must be locked unless a staff member is on the premises.ACCESS ‡ Understanding of the safety practices. ± Written form ± Sighted by the staff member designated to be present in the laboratory.

‡ 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 ‡ No visitors into the laboratories without permission.

but shall remain closed when in use. ‡ Doors displaying biohazard symbol shall not be propped open. .ACCESS ‡ The universal biohazard symbol displayed at specific laboratories in which manipulations of organisms with moderate and heavy risk.

PERSONAL PROTECTIVE EQUIPMENT and PROCEDURES ‡ ‡ ‡ ‡ ‡ ‡ Gowns Safety Glasses and Eye Protection Gloves Mask Hand washing High Risk Individuals /Antenatal Considerations .

‡ Soiled gowns in excess of "normal use" are to be removed from circulation. ‡ Students wear gowns at all times in the laboratory. . ‡ Laundered every fortnight during semester.Gowns\Coats ‡ Rear opening wrap around gowns. ‡ 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 .

. ± If open cuts or skin conditions. ± Mopping up a spill. please notify so that stocks can be replenished. not essential. If you use the last gloves. ± Where there is a high risk of contaminating hands. ± When instructed by the demonstrator ± Adequate supplies of gloves. ‡ Worn under the following circumstances.Gloves ‡ For routine work.

Safety Glasses and Eye Protection ‡ ³Safety glasses" for all practical sessions. ± ³Fresh" sterile distilled water ± Replace every month ± Record date of "renewal" . ± Risk of splashing with infectious or corrosive liquids. ± When opening the autoclave. ‡ An eyewash station.

‡ The "Hibiclens" dispensers at all hand washing sinks within the laboratory must always be sufficiently full. ± Must before leaving the laboratory. .Hand washing ‡ The standard hand washing procedure is to use running water and "Hibiclens".


Hand washing 35 .

‡ Student with medical condition.High Risk Individuals /Antenatal Considerations ‡ Immunocompromised or particularly susceptible to infection need to take additional precautions. ± Pregnant women. ‡ Children. so any female student or staff member should discuss the matter with the academic in charge. .

Chemical Safety ‡ ‡ ‡ ‡ ‡ ‡ Do not inhale the fumes. Wash hands. Wear goggles Lab apron Never taste any chemicals (never taste anything). Never pour water into a concentrated acid. .

‡ hands dry before using electrical equipment. . ‡ Unplug cords by pulling the plug and not the cord.Electrical Safety ‡ Lay electrical cords where no one can trip on them or get caught in them. ‡ Unplug all electrical equipment at the end of the lab period. ‡ Never poke anything into electrical outlets.

‡ Dry glassware should be heated. ‡ Heating a test tube. move it around slowly. . ‡ Use tongs and/or protective gloves. ‡ Never reach across an open flame. ‡ Do not hold it in your hand ‡ Never leave a burner or hotplate unattended.Heating Safety ‡ Let burners and hotplates cool down before touching. ‡ Wait until the striker is in place before you turn on the gas.

LABORATORY EQUIPMENT ‡ Autoclave ‡ Biohazard Cabinet ‡ Gas Supply .

‡ Heat safety gloves. ‡ Students under the direct supervision. ‡ Students must be given instruction before using. ‡ Safety glasses ‡ De-pressurized before opening. .Autoclave ‡ To avoid Noxious fumes turn on the extraction fan.

± Wipe down the interior work surfaces. ± Close doors to minimize the disruption of the airflow. . ± allow it to run motor for 30 sec. ± Check annually by Laminar Airflow Services. ± Either install the shield or leave the room if the UV lamp is required. ± Notify about any major spills.Biohazard Cabinet ‡ Class II cabinet to protect the operator.

Pipetting ‡ The significant and consistent cause of occupational infections. . ± Avoid rapid mixing of liquids by alternate suction and expulsion. ± Don¶t forcibly expel material from a pipette. ± Prefer pipettes that do not require expulsion of last drop of liquid. ± Don¶t bubble air through liquids. ± Reduce creating aerosols. ± Plug pipettes with cotton.

‡ Selection depend upon ease of operation and type of work.Pipetting ‡ Drop material close to the fluid or agar level. ‡ Place contaminated pipettes in a container having suitable disinfectant immersed. ‡ Do not place contaminated pipettes on the bench top. .

‡ The gas supply is to remain off until a staff member declares it safe to switch it back on. fire or back burning of an individual Bunsen burner. ‡ A master isolation switch to the entire laboratory. deactivate the isolation switch immediately. . should be de-activated at the end of each day. ‡ Incase of a spill of flammable liquid.Gas Supply ‡ Each room has an isolation switch.

. ‡ The secondary container sealable and non breakable.Transport ‡ When viable organisms ± Cultures ± Specimens ± stock organisms are transported to or from the laboratory. the "primary" container "secondary" container.

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.

‡ Vigorous shaking of liquids & when sealed ampoule containing a lyophilized or liquid culture is opened. handle with care. ‡ Wear coat & gloves.Opening containers ‡ Well-lighted and designated areas. ‡ absorbent paper towel to: ± facilitate clean-up ± reduce generation of aerosols ‡ Leaking or broken Specimens in BSC. ‡ Tubes containing bacterial cultures. 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 ‡ ³Yellow" sharps container at the back of each laboratory. ‡ Do not wander around the laboratory carrying sharps. ± Needles ± scalpel blades ± other sharp materials ‡ Always move the "sharps container" to your work place to dispose of such items. .

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 ‡ Containers of general purpose phenolic.

non contaminated paper or matches.Biogram Buckets ‡ Disposing pipettes. or if grossly soiled. ‡ 1/40 dilution of the commercial product 18% phenol equivalent. place tip first into the biogram ‡ Not for Gram stains. . ‡ Change weekly.

‡ 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".Biogram Buckets ‡ autoclaved prior to removing or disposing of the contents. .

e. ‡ Do NOT use these bins for ± Paper towel discarded after hand washing ± Blotting paper discarded after blotting slides. ± Used culture plates ± Contaminated paper towel. ‡ autoclaved before disposal.Biohazard Bin ‡ Centre of each laboratory. ‡ For the disposal of contaminated waste. ‡ Left open to allow penetration of the steam . ‡ Not to be used sharps & Any non contaminated paper wastes.g.

‡ Container 1: ± Contaminated and non contaminated recyclable glass or plastic tubes.Billy Cans ‡ Two stainless steel tins at the front of each laboratory. . ± 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 towel from hand washing (but not bench wiping) ± Blotting paper from blotting Gram stains.g. e..Paper Bin ‡ To be used for non contaminated paper waste only.

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.Broken Glass ‡ When any glassware is broken. .

‡ Don¶t dispose of any solid material. .Sink ‡ Don¶t place any hazardous or infectious materials.

‡ No set of instructions ‡ sound microbiological judgment in their management.CLEAN-UP PROCEDURE FOR BIOHAZARD SPILLS ‡ Spills involve infectious materials. are complex. ‡ 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. ‡ General purpose laboratory disinfectant (Biogram) can be used . ± airborne particles (aerosols) ‡ airborne particles pose the greatest risk. ± Splashes. remain airborne and be dispersed to other areas.

. ‡ 8. 5000 ppm of available chlorine.Diversol ‡ ‡ ‡ ‡ Spills involving human blood or body fluids. Limited shelf life. Sachets stored on the chemicals shelf in the preparation room.3 Squirt Bottles ‡ Sufficient "biogram squirt bottles". ‡ Clearly labelled.

. ± The biological nature. ± The physical nature ± The volume.Assessing a Biohazard Spill ‡ The designation into ± "minor" ± "major" ‡ The supervising staff member responsible.

first wash hands. ‡ Put on gloves. ‡ ‡ ‡ ‡ ‡ . Low potential for generation of aerosols. Remove and replace any contaminated protective clothing. If hands contaminated.Minor biohazard spills Minimally hazardous material. ‡ 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. ‡ Remove gloves and wash hands. . ‡ Wipe over general area again with paper towel dampened with disinfectant.

‡ Hold breath ‡ Warn others ‡ Leave the room immediately. ‡ Notify MSO (preferably) or other senior. ‡ Remove any contaminated clothing and wash any contaminated body surfaces. ‡ Close doors and place a "DO NOT ENTER" sign on the door.Major biohazard spills ‡ Major risk with larger volume ‡ Considerable production of splashes and aerosols. .

likely to have been contaminated. ‡ Disinfectant areas around the spill. face masks and safety glasses" before entering the spill area. gloves. . ‡ Do not pour disinfectant directly onto the spill ‡ Lay paper towels wetted with disinfectant onto the spill for 30 min.Major biohazard spills ‡ "Spill Clean Up Team" ± two to clean up ± one to supervise. ‡ "gowns. ‡ Re-enter after 30 min.

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: What To Do: Burns Immediately flush with cold water until burning sensation is lessened.

bruises What To Do: ‡ Do not touch without gloves.First Aid Injury: Cuts. . ‡ 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.

‡ If a foreign object is lodged in the eye.First Aid Injury: Eyes What To Do: ‡ Flush eyes immediately with plenty of water for several minutes. 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. .

apply vinegar or boric acid. For base spills.First Aid Injury: Spills on the skin What To Do: Flush with large quantities of water. apply baking soda solution. For acid spills. .

‡ Remove wire with rubber gloves. .First Aid Injury: Electrical shock What To Do: ‡ Shut off the current at the source. ‡ Alert the teacher immediately.

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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!!!! .

Microbiological safety. tm WHO Microbiology Rules for Using Disinfectants eHow com. Page Kennedy Middle School 2002-2003 Presented by: Leonard LaFazia .Reference ‡ ‡ ‡ ‡ ‡ ‡ Food Standards Agency . February 2000 Mrs. htmL Microbiology Safety and Staff Induction Manual University of Tasmania Launceston Campus Version 2. html Guidelines on Standard Operating Procedures for MICROBIOLOGY Safety in Laboratories.