Laboratory Health and Safety IMBB 2017 RAB, Kigali - Rwanda May 02 13, 2017 Roger Pelle
Laboratory Safety WHY? Reduce or eliminate exposure to potentially hazardous agents of Workers Products Co-workers, lab support personnel Environment HOW? By the combination of Laboratory practices and procedures Laboratory facilities Safety equipment These allow the <containment> of biohazards
Safety in the Laboratory
Know Your Environment and Manage Risks Outline the fundamentals of lab Practice. Describe lab operations (field and lab), that require lab specific safety guidelines. Describe the various types of lab hazards and their containment protocols. Describe the bio-safety levels to which pathogens are assigned. Describe the universal bio-safety precautions required during specimen collection, packaging and shipment. Describe the relevance of safety programs in institutions.
Control Measures Determine hazards and evaluate risks using relevant available data Determine and use control measures needed to minimise those risks; e.g.: 1. Use a less risky substance 2. Use a safer form of that substance (e.g. solution instead of powder; GelRed vs EtBr) 3. Totally enclose the process (e.g. a glove-box) 4. Partially enclose the process (e.g. with a fume cupboard) 5. Ensure good general ventilation
Control Measures (Cont ) 6. Safe systems of work 7. Reduce exposure times, increase distance, reduce volumes 8. Personal protective equipment Wear the clothing and protective wear identified in your risk assessment Laboratory coats must be kept fastened Don t wear sandals or open shoes Long hair must be tied back Use the correct protective gloves according to materials, substances or chemicals used Remove your gloves before using instruments, telephone, and leaving the laboratory
Laboratory Hygiene Never eat, drink or smoke in a laboratory Never apply cosmetics Never touch your face, mouth or eyes Never suck pens or chew pencils Always wash your hands before you leave and especially before eating
Protecting your health If you have an allergy to lab materials or suffer from a medical condition which may affect you in the laboratory (eg diabetes or epilepsy), ensure that your supervisor knows
First Aid All laboratory workers should undergo simple first aid training For ALL chemical splashes, wash with plenty of water for 10 minutes Control bleeding with direct pressure, avoiding any foreign bodies such as glass Report all accidents to your supervisor or departmental safety officer
Safety in the Laboratory 1. Eating, drinking, smoking and applying cosmetics prohibited. 2. Mouth pipetting is prohibited. 3. Staff must behalf in a safe and responsible manner at all times. 4. Appropriate protective clothing must be worn at all times. 5. The laboratory must be kept clean and tidy and should contain items necessary for the work carried out. 6. Appropriately decontaminate all working surface at the end of each working day and after any spillages. 7. Staff must wash hands when leaving the laboratory.
Safety in the Laboratory (2) 8. Avoid the formation of aerosols or the splashing of materials. 9. Appropriately decontaminate all contaminated waste or reusable materials before disposal or reuse. 10. Access must be restricted to authorized personnel only. 11. Report all incidents and/or accidents immediately and appropriate action taken to prevent further occurrences. 12. All staff must be adequately trained, both in the duties they perform and in all safety aspects of laboratory work.
Equipment That Prevent Hazards Bio-safety cabinets Pipetting aids Centrifuge caps Plugged plastic pipettes Plugged pipette tips Hard top foot ware gum boots Wall shackles for gas cylinders and metal cabinets for corrosive materials Disinfectant spray bottles
Laboratory Design That Prevent Hazards Laboratory room - adequate Laboratory floor non polished/ nor carpeted Proper lighting and ventilation Benches standard height and depth Sink free flowing water and elbow tap openers Drawers and cupboards
Laboratory Hazards
What are the general hazards in a laboratory? 1. Fire 2. Breakage of glassware 3. Sharps 4. Spillages 5. Pressure equipment & gas cylinders 6. Extremes of heat & cold 7. Chemical hazards 8. Biological hazards 9. Radiation 10. Electrical 11. Mechanical
Laboratory Associated Hazards Acquired laboratory infections Burns Cuts Pricks. Use of infected sharps and needles Dangerous equipment Faulty electrical appliances Uncertified working facilities and equipment Falling freestanding appliances Inadequately trained personnel handling infectious materials
Sources of Biological Hazards How: Accidental pricks/cuts Contamination of open cuts Inhalation or splashes to eyes, mouth and onto clothing Ingestion Spills on bench tops, floors Contaminated utensils door knobs, spray bottles Specimens, cultures, storage and shipping equipment
Sources of Chemical Hazards Chemical disinfectants Laboratory reagents
Avoiding Fires Flammable substances Use minimum quantity Store in special storage cabinet Use temperature-controlled heating sources (e.g. water-bath rather than hotplate or Bunsen burner) Keep fire doors and Lab doors shut at all time Make sure that you know what to do: If you have a fire If you hear a fire alarm
Glassware Use correct techniques for the insertion of tubing onto glassware Never use glassware under pressure or vacuum unless it is designed for the job and suitably shielded Dispose of chipped or broken glassware it is a risk to you and others Always dispose of broken glass in a glass bin or sharps bin and not in a general waste bin
Spillages Clear up spillage promptly You will already have determined how to do this as part of your risk assessment Dispose of any hazardous material as toxic waste Messy workers are usually poor workers
Gas Cylinders Never use without formal training Minimise the number in a laboratory Store externally whenever possible Cylinders are heavy and can do serious damage to you if they fall Ensure that they are chained when in use Move only with a cylinder trolley Use regulators & control equipment suitable for the gas concerned Consider the consequences if your cylinder leaks
Cryogenics Liquid gasses are extremely cold and can cause burns Liquid gases evaporate and many can cause asphyxiation If you need to take cryogens in a lift, there are special procedures to follow You must have special training to use them
EQUIPMENT THAT MAY CAUSE HAZARDS Centrifuges Ultra-sonicators Refrigerators and freezers Cold storage tanks (including liquid nitrogen) Vortex mixers Gas cylinders Gels and power sources UV sources, microwave ovens, autoclaves Scalpel, Etc.
SOURCES OF ELECTRICAL HAZARDS Improper insulation Overloading power points Use of wrong gadgets Power surge
Use of Electrical Equipment Always do a visual check on electrical equipment before use, looking for obvious wear or defects All portable electrical equipment must have a current PAT test sticker Never use defective equipment NB: Portable appliance testing (PAT) is the term used to describe the examination of electrical appliances and equipment to ensure they are safe to use
Use of Laboratory Equipment Never use any laboratory equipment unless you are trained & have been authorised to do so As well as injuring yourself you may cause very costly damage
General Lab Tidiness Keep your workplace tidy Clear up waste, deal with washing up and put things away as you finish with them Make sure everything is safe before you leave things unattended A tidy laboratory avoids accidents to everyone X
SOURCES OF PHYSICAL/MECHANICAL HAZARDS Floor polished Over crowding Improper use of equipment Improper furniture Improper storage of chemicals and reagents and carry-on appliances, e.g. cool boxes so they fall out of their storage sites and injure operators
SOURCES OF RADIATION HAZARDS Use of radioisotopes Improper disposal Improper storage Improper use of ultra violet lamps
Waste Materials Part of your risk assessment will be to determine how to dispose of waste lab materials safely Solvents and oils must be segregated into the correct waste bottle or drum Your department will help you determine what to do with chemical or biological materials Do not put materials down the drain or in with normal waste unless authorised to do so
Working outside normal hours and at weekends You will need to attend training courses and have permission from your Head of Department before working outside normal hours Most experimental work is not permitted Your supervisor will explain the requirements in more detail
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!!!!
Biological Safety and Containment
Biosafety and dangerous goods Infectious biological agents classified as dangerous or hazardous goods: An agent of biological origin that has the capacity to produce deleterious effects on humans: i.e. microorganisms, toxins and allergens derived from these organisms
Biosafety levels Risk group 1: well characterized agents not known to cause disease in healthy human adults and of minimal potential hazard for personnel and environment. e.g.: Bacillus subtilis, lactobacillus, etc.. Risk group 2: agents of moderate potential hazard for personnel and environment, low probability of transmission, available treatment or prohylaxis ex: Klebsielle, measles virus, toxoplasma spp, hepatitis B, etc. Risk group 3: agents which may cause serious or potentially lethal disease but with no transmission by aerosol (HIV) or if with available treatment (Mycobacterium tuberculosis) Risk group 4: High risk for humans and environment, no treatment or prophylaxis eg Ebola, Marburg, etc.
Risk assessment Risk: probability of harm, injury of disease to occur In laboratory, assessment of risk focuses on prevention of laboratory acquired infections In order to reduce to absolute minimum the risk of exposure to an agent.
Factors of interest in risk assessment for biosafety consideration in laboratory work. 1. Pathogenicity 2. Route of transmission 3. Agent stability 4. Infectious dose 5. Concentration 6. Origin 7. Available data 8. Potential of secondary spread in the community 9. Availability of prophylatic or therapeutic agents. 10. Experience and skill of at risk personnel.
Guidelines for waste disposal All potentially contaminated materials must be treated to render them non-infectious prior to disposal. Biological waste management program should include the following elements:- (i) Segregation of infectious waste from other waste (ii) Packaging of infectious waste to prevent leaks and spills. (iii) Decontamination of infectious waste (iv)disposal of infectious waste (v) Contingency plans for emergency.
DISPOSAL PROTOCOLS Contaminated air Bio-safety cabinets Contaminated solid waste Autoclaving and incineration. Contaminated sharps Disinfection and incineration. Non-contaminated solid waste Burning Contaminated liquid waste Disinfection and autoclaving. Chemical liquids Neutralization, dilution
BSL-1 Practices (1) 1. Eating, drinking, smoking and applying cosmetics prohibited. 2. Mouth pipetting is prohibited. 3. Staff must behave in a safe and responsible manner at all times. 4. Appropriate protective clothing must be worn at all times. 5. The laboratory must be kept clean and tidy and should contain items necessary for the work carried out. 6. Appropriately decontaminate all working surfaces at the end of each working day and after any spillages. 7. Staff must wash hands when leaving the laboratory.
BSL-1 Practices (2) 8. Avoid the formation of aerosols or the splashing of materials. 9. Appropriately decontaminate all contaminated waste or reusable materials before disposal or reuse. 10.Access must be restricted to authorized personnel only. 11. Report all incidents and/or accidents immediately and appropriately action taken to prevent further occurrences. 12. All staff must be adequately trained, both in the duties they perform and in all safety aspects of laboratory work. 13. Maintain an insect/rodent control program.
BSL-1: Primary containment Personnel protective gear Lab coat, gowns, gloves, Eye and face protection, Safety equipment (primary barriers) Pipetting devices
BSL-1: Secondary barriers (facility design and construction) Laboratories with doors Windows with flyscreen Sink for handwashing Work surface easily cleaned Bench tops impervious to water Robust furnitures
BSL-2: Microbiological practices Standard Microbiologic practices: As in BSL-1 Special Microbiologic practices: Needles and sharps precautions+++ Don t break, bend or re-sheath or re-use syringe and needle Use sharp containers, with disinfectant, then incinerate. Don t handle broken glass with bare hands Use polypropylene/plastic wares. Use leak proof autoclavable waste containers for discard. Follow policy on lab entry procedures Limit lab access to immunocompromised individuals Provide bio-safety manual and training updates to lab operators.
BSL-2: Primary containment As for BSL-1 + Personnel protective equipment Mask, respiratory protection Safety equipment Use of class II Biosafety cabinets for work with infectious agents involving: Aerosols and splashes Large volumes High concentrations Safe centrifuge and rotors Eyewash available
BSL-2: Secondary barriers: (facility design and construction) Normal construction Lockable doors with restricted access Bench top impervious to water Adequate illumination Foot or elbow operated sink for hand-washing Biosafety cabinet Eyewash available Autoclave available
BSL-3: Microbiological practices Standard Microbiologic practices: as in BSL-1 and 2 Special Microbiologic practices: as in BSL-1 and 2 + Restricted access upon authorisation of the supervisor, limit entry to immuno-compromised persons. Appropriate training Strict following of guidelines and procedures Medical surveillance, including vaccinations. Report incidents/accidents Work controlled in Biosafety Cabinet, 2x2 rule. Use of bio-aerosol containing equipment (centrifuge, Prompt decontamination of spills Treatment of effluents and waste
BSL-3: Primary containment Personnel protective equipment as in BSL-1 and 2 + Special lab gowns Double gloves, Mask, respiratory protection, Safety equipment as in BSL-1 and 2 + Use of certified class IIB Biosafety cabinets Air filtration and unidirectional flow
BSL-3 Secondary containment or barriers: facility design and construction As in BSL-1 and 2+ Separated building or isolated zone Double door entry Directional inward airflow Single way pass air Negative pressure gradient to most risk areas Room penetration sealed Wall, floors and ceilings are impervious and water resistant for easy cleaning Autoclave double door
BSL-4:Microbiological practices Standard Microbiologic practices: As in BSL-1-3 Special Microbiologic practices: As in BSL-1-3 + Controlled access as per guidelines and supervisor instructions through a changing room (s). Complete change of clothing before entry in the laboratory and on exit; clothing autoclaved after single use. Decontamination shower at each exit Personnel shower at each exit Chemical decontamination, autoclaving and incineration of all waste. Chemical decontamination and autoclaving of all effluents Enter all materials through double door autoclave or fumigation pass box chamber which is air-locked.
BSL-4: Primary containment Personnel protective equipment as in BSL-1-3 + Specific one single use underclothing and one piece suit with gloves, hats Boot rims and sockets fixed by tapes. One piece positive pressure personnel suit ventilated by a life support system (BSL4) (or work in certified class 11B Biosafety cabinets) Safety equipment as in BSL-1-3 +
BSL-4 Secondary containment or barriers: facility design and construction (1) As in BSL-1-3 + Separate building or separate zone Double door entry Directional inward airflow Single pass air Double HEPA filter on supply and exhaust air Room penetration sealed Walls, floors and ceilings are sealed to form an internal seal
BSL-4 Secondary containment or barriers: facility design and construction (2) As in BSL-1-3 + Connecting inner and outer doors interlocked Double door autoclaves with validated cycles Validated double tank for thermic decontamination of effluents Communication system Emergency breathing air Emergency generator Emergency exist Alarm systems on all biosefy related devices
Biological safety cabinets (1) Protection is provided by: Directional airflow HEPA filters: high efficiency particulate air filters HEPA filters traps only particulates 0.3µ Chemicals, fumes, vapors passed through may damage HEPA filters.
Biological safety cabinets (2) Location: Isolated from other work areas Removed traffic ways and doors Far from airflow
Biological safety cabinets (3) Operating procedures Turn BSC Load BSC with all needed supplies: materials must be well within the cabinet Check inward flow Place discard plan within the cabinet After entering the hands in the cabinet wait 1 minute for stabilizing of the flow and move slowly the arms in the cabinet Avoid movements around the BSC At the end of the work decontaminate all the items before taking them out. Decontaminate interior of the BSC Let in run 10 to 15 minutes Shut off and switch on the UV lamp
Bio-Safety Cabinets (BSC) Classification Three general types available Class I Class II Class III
Class I Negative Pressure ventilated cabinet Designed for general microbiological low and moderate risk agents e.g. Staphylococcus Inward flow of unfiltered air from the laboratory
CLASS-1 BSC Protection of worker by inward airflow Protection of environment by exhaust to outside (with HEPA filter) No protection of the manipulation
CLASS II Designed with inward air flow to protect personnel. HEPA-filtered downward laminar flow for product protection. HEPA-filtered exhaust air for environment protection. Certified at installation, any time moved and annually. Designed for research laboratories. The cabinet should be used by trained personnel to ensure proper use. The cabinets should be located away from traffic patterns and doors.
Class III cabinet (1) Designed with Glove holes (3) and a left side glove hole to the materials entry port. The port has a pressurised sealed door for materials. It has another sealed door opened from inside using the leftmost glove hand to let in materials and is then sealed. HEPA filtered Air flows into the cabinet work area only inside cabinet. Operator works on materials through the gloves and has no contact with air inside the cabinet. Hole into the dank allows materials to be dropped into decontaminant in the dank and taken out through a sealed dank door without loosing the air-flow pressure.
Class III cabinet (2) HEPA-filtered exhaust air for environment protection. Certified at installation, any time moved and annually. Designed for use with level 4 pathogens that would infect man directly. The cabinet should be used only by trained personnel to ensure proper use. The cabinets should be located inside a level -3 laboratory.
Establishment of Bio-safety programmes Every laboratory must have a written biological safety program. It should include the following:- 1. A policy statement signed by the laboratory director, and appointment of a bio-safety officer. 2. Guidelines for appropriate biological safety practice, storage and shipment of materials in accordance with WHO regulations. 3. Waste disposal Protocols. 4. An employee education and training program.
Medical surveillance The requirements for medical surveillance must be developed and implemented for all employees potentially exposed to biological agents. This provides a means for early detection of laboratory acquired infections. It helps assess the efficacy of protective equipment and procedures.
Thank you IMBB 2017 RAB, Kigali - Rwanda May 02 13, 2017 Roger Pelle r.pelle@cgiar.org