BIOLOGICAL SAFETY Azian Harun
MBBS, M.Path (Microbiology), PhD
Department of Medical Microbiology & Parasitology School of Medical Sciences
Universiti Sains Malaysia
Content
• Introduction & definitions
• Legislation matter
• Biological materials & hazards
• Principles of containment
• Standard microbiological practice
• Guidelines & responsibilities
• Resources
Introduction
• Bio-related research activities may involve manipulation of microbial, animal or plant cells.
• The risks associated with these activities arise from the samples and /or the procedural requirements.
• Adherence to standard microbiological techniques and using facilities suitable to the risk level of the pathogen helps to protect the researcher from laboratory-acquired infections.
Definition
• Biosafety - the containment principles, technologies and practices that are implemented to prevent the unintentional exposure to biological agents and toxins, or their accidental release
(WHO/CDS/EPR/2006.6) Keeping you and others safe from biological hazards and
meeting statutory requirements
To protect:
• Workers & co-workers,
• Products
• Environment
• Students
• Visitors
Why biosafety practices?
What legislation applies?
LAW OF MALAYSIA
ACT 678
Biosafety Act 2007
Hazard Categories
• Chemical: corrosive, toxins, carcinogens, mutagens, flammables, explosive
• Physical: ionizing and non-ionizing radiation, noise, temperature extremes, electrical mechanical
• Biological: pathogens, organism, recombinant DNA, toxins produced by organisms and human or animal tissue, body secretions, & blood
Biological materials & hazards • Hazards related to bio research
can be classified into : – hazards related with the
pathogen or human/animal cells being used in research.
– related with the procedures and practices followed in the lab.
All organisms (especially animals and humans) are incubators for disease-causing organisms.
Biological hazards
Virus Bacteria /
Toxins Fungi Parasites Prions
Adapted from: Barry Byrne, UCD
Organism must get onto/into body in sufficient amount and begin to grow.
Mechanisms: Ingestion
Inhalation – aerosol
Puncture wounds –needles/glass ware.
Direct contact
Mucous membranes – especially eyes and nose
Biological materials & hazards - mechanisms of introduction
Biological materials & hazards
Hazardous Characteristics of microorganisms
• Risk group I
• Risk group 2
• Risk group 3
• Risk group 4
Classification of Infectious Microorganisms by Risk Group
https://www.cdc.gov/biosafety/publications/bmbl5/BMBL5_sect_II.pdf
Biological materials & hazards
• Other hazardous characteristics to consider: – probable routes of transmission of laboratory
infection
– infective dose
– stability in the environment
– host range
– endemic nature
Principles of containment
• The term containment is used to describe the safe work practices in handling infectious agents to reduce exposure to laboratory personnel and others.
Principles of containment – Biosafety levels
• It consists of a combination of laboratory practices,equipment and facilities suitable to the procedures being performed and hazards of the pathogen.
• Biosafety levels corresponds to organism risk groups (RG).
• A lower risk group can be assigned a higher biosafety level, if required based on biological risk assessment.
Principles of containment
• 4 Biosafety Levels (BSL) – BSL 1 – BSL 2 – BSL 3 – BSL 4
• Biosafety levels define the lab requirements, protective clothing, and work practices.
Biosafety Level 1
BSL 1 – Organisms not known to cause disease in healthy
human adults
– Pose minimal hazards under ordinary conditions of handling
– E.g. S.cerevisiae, non-pathogenic E.coli
CDC BMBL 5th Edition 2009
Biosafety Level 1 • Standard practices
– Use mechanical pipetting devices – No eating, drinking, smoking in lab – Minimize splashes and aerosols – Decontaminate work surfaces – Safe handling of sharps – Wash hands before leaving lab – Biohazard sign is posted
Biosafety Level 1
• Protective Clothing – Lab Coat
– Gloves
– Eye Protection (if splashes are likely)
BioSafety Level 2
• BSL 2 – Work with well-characterized agents not known to
cause disease in healthy adult humans; minimal hazard to laboratory personnel and the environment
– Examples: Salmonella Hepatitis B Virus Measles Virus (not aerosolized)
CDC BMBL 5th Edition 2009
Biosafety level 2
• Other materials handled at BSL 2 (Requires compliance with OSHA Standard)
– Human blood – Human tissues – Human cell lines – Biotoxins – Viral vectors
Biosafety Level 2
• Standard practices – All requirements for BSL 1 plus:
• Access to laboratory is limited or restricted when work is being conducted; door is closed
• Personnel have specific training in handling pathogenic agents
• Extreme precautions are taken with contaminated sharp items
Biosafety Level 2
• Standard practices cont. • Written Biosafety Procedures
• Report spills
• Minimize aerosol generation
• Personnel receive immunizations or testing (e.g. Hep B vaccine or TB skin Testing)
Biosafety Level 2
• Personal Protective Equipment
– Lab Coats
– Gloves
• Double Gloving when appropriate
– Eye Protection - Safety Glasses/Shield
– Biosafety cabinets or sealed safety centrifuge
BioSafety Level 3
• BSL 3 – Use of indigenous or exotic agents which may
cause serious or potentially lethal disease from exposure by the inhalation route.
• Examples of organisms: – Mycobacterium tuberculosis
– Hantavirus
– SARS
CDC BMBL 5th Edition 2009
Biosafety Level 3
• Standard Practices
–All requirements for BSL 2 plus: • Very limited lab access
• 2 doors in a series to access lab
• Able to decontaminate entire lab
• Special exhaust ventilation
(Not re-circulated, no airflow outward)
Biosafety Level 3
• Standard Practices –cont. – High level of training
– Personnel receive vaccinations if available
– Work in Biosafety cabinets and/or respirator used
Biosafety Level 4
• BSL 4 – dangerous and exotic agents with a high risk of
aerosol-transmitted laboratory infections and life-threatening disease.
• Examples – Ebola Virus
– Monkey B Virus
– Marburg Virus
CDC BMBL 5th Edition 2009
BioSafety Level 4
• Requirements – All requirements for BSL3 plus:
• Class III Biosafety cabinet or positive pressure suits
• Shower/Change rooms
• Clothing autoclaved before laundering
• Air locks
• All waste contained and decontaminated onsite
• Space suits
Space Suit
Standard microbiological practice
• Safe work practices for all levels
• Include all staff/researchers – NO EXEMPTION!
• Knowledge, attitude, practices, responsibilities
Safe Work Practices for all Levels
• Wash hands after work; when removing gloves; before leaving lab
• No eating, drinking, applying cosmetics, handling contact lenses in the lab.
• Maintain labs in clean, orderly fashion.
• Disinfectants & decontamination
Safe Work Practices for all Levels
• Limit access to lab when work with organisms is in progress
• Use good microbiological techniques
• No mouth pipetting
• When possible use plastic instead of glass
Sharps safety
• Sharps include needles, syringes, razor blades, lancets, slides, scalpels, pipettes, micropipettes, pipette tips, broken plastic or glassware, and other devices capable of cutting or piercing the skin.
Sharps safety
• Contaminated needles are not bent, recapped, or removed.
• If recapping is required, use a mechanical device or a one handed technique.
Physical containment
• BioSafety Cabinets (BSCs) – Protect personnel and the environment from
hazardous particulates and from infectious agents
• Laminar Flow Clean Benches (LFBs) – Non Hazardous work only
– Protect work from contamination
– Do not protect personnel
Laminar Flow Cabinets
• Laminar flow cabinets are designed to provide a clean-air environment to protect the work
• They DO NOT provide protection for the operator
BioSafety Cabinets (BSC)
• 3 BSC Classes
• All exhaust is HEPA filtered before leaving
the cabinet
Biological Safety Cabinets • Class 1
– Some protection to user & environment – No protection to work
• Class 2 – Good protection to user, the
environment and the work • Class 3
– Total enclosure You must choose the correct safety cabinet for the
job you are doing. Seek advice from your Biological Safety Officer
Blood & bodily fluids • Use screened blood • All blood and bodily fluids should
be considered potentially infectious and treated with due care
• Injuries involving body fluids must be reported to your supervisor IMMEDIATELY
Spillages • Report
• Wear protective clothing
• Clean up
• Decontaminate – ensure that you always have material available to do so
• Ensure waste is disposed of properly
You spill = you clean up!
Autoclaves
• Potentially dangerous equipment
• Require regular maintenance and an annual statutory examination
• Discard autoclaves must be calibrated every 6 months
• You must be trained to use them
Disposal of Waste
• Make sure you follow the University policy carefully
• Ask your supervisor or a senior member of technical staff if you do not know the correct disposal method for the materials with which you are working
Sharps Disposal
• Sharps containers for disposal of these items are conveniently located and easily accessible.
Sharps Disposal
• Syringes and syringes without a needle attached go into a sharps container
• Contaminated micropipettes, pipette tips, and Pasteur pipettes are discarded in a puncture-resistant container or a sharps container for disposal.
• Include broken glassware, glass slides & cover slips
Sharps Disposal
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!!!!
Guidelines & responsibilities
• Standard Operating Procedures (SOPs)
• Refer to available SOPs/guidelines in your lab or institution.
• Know your Safety Officer/Biosafety Officer in charge
• Responsibilities of employers • Responsibilities of
workers/staff/users
