Transcript Biosafety Level 2: . Special Practices

General Biosafety
Tassanee Eamkamon
Biosafety Professional
Armed Forces Research Institute of Medical Sciences
[email protected]
23 March 2010
Objectives
•
•
•
Principle of Biosafety
Overview of Biosafety Levels (BSL) and Risk
Groups (RG)
Biosafty Level 2 Requirement
2
Principle of Biosafety
• A fundamental objective of any biosafety program is the
containment of potentially harmful biological agents.
• The purpose of containment is to reduce or eliminate
exposure of follows to potentially hazardous agents.
- Laboratory workers
- Other persons,
- Outside environment
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Principles of Biosafety
• The term "containment" is
used in describing:
- Safe methods (Practices)
- Safety Equipment (Primary Barriers and
Personal Protective Equipment)
- Facilities Design and Construction
(Secondary Barrier)
www.pandemicfluonline.com/%3Fp%3D1359
4
Overview of Biosafety Levels (BSL) and Risk Groups (RG)
Biosafety Level?
CDC-NIH
http://www.cdc.gov/OD/OHS/biosfty/bmbl5/BMBL_5th_Edition.pdf
Overview of Biosafety Levels (BSL) and Risk Groups (RG)
Biosafety Level (BSL)
Safe Practice
Safety
Equipment
Facility
Design
• Combination
• 4 Levels
• Each level is specifically appropriate
for:
- Operations performed
- Risk Group and documented or suspected
routes of transmission of the infectious
agents
6
BSL 1
Safe
Practice
Safety
Equipme
nt
Facility
Design
Appropriate for:
 Undergraduate and secondary educational training and
teaching laboratories
 Laboratories in which work is done with defined and
characterized strains of viable microorganisms not known
to consistently cause disease in normal, healthy humans
- Bacillus subtilis
- Vaccine strains that have undergone multiple in vivo passages
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BSL 2
Safe
Practice
Safety
Equipme
nt
Facility
Design
Appropriate for:
 Clinical, diagnostic, teaching, and other laboratories
 Lab work with a broad spectrum of indigenous moderaterisk agents that are present in the community and associated
with human disease of varying severity
 Lab work with any human-derived blood, body fluids,
tissues, or primary human cell lines where the presence of an
infectious agent may be unknown
-Hepatitis B virus
-Salmonellae
- P. faciparum
-H1N1
BSL 3
Safe
Practice
Safety
Equipme
nt
Facility
Design
Appropriate for:
 Clinical, diagnostic, teaching, research, or
production facilities
 Lab work with indigenous or exotic agents:
-with a known potential for aerosol transmission
-may cause serious and potentially lethal infection
-Mycobacterium tuberculosis
-Chikungunya
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BSL 4
Safe
Practice
Safety
Equipme
nt
Facility
Design
Appropriate for:
 Lab work with dangerous and exotic agents which:
-- pose a high individual risk of life-threatening disease
-- may be transmitted via the aerosol route
-- there is no available vaccine or therapy
-Ebola virus
- Hendra and Nipah virus
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Overview of Biosafety Levels (BSL) and Risk Groups (RG)
Risk Group?
Overview of Biosafety Levels (BSL) and Risk Groups (RG)
Risk Group (RG)
Risk classification based on:
Pathogenicity
Modes of transmission
Host range
Availability of effective preventive measures
Availability of effective treatment
Risk Group (RG)
1. WHO (2004)
2. NIH Recombinant DNA Guidelines (USA, 2002)
3. Australian/New Zealand Standard (2002)
4. Canadian Laboratory Safety Guidelines (2004)
5. European Economic Community (2000)
Overview of Biosafety Levels (BSL) and Risk Groups (RG)
RISK GROUP
NIH GUIDELINES FOR RESEARCH
CLASSIFICATION INVOLVING RECOMBINANT DNA
MOLECULES 2002
WORLD HEALTH ORGANIZATION
LABORATORY BIOSAFETY MANUAL
3RD EDITION 2004
Risk Group 1
A microorganism that is unlikely to cause
human or animal disease.
(No or low individual and community risk)
Agents that are not associated with
disease in healthy adult humans.
Bacillus subtilis or Bacillus licheniformis
Risk Group 2
Agents that are associated with human
disease which is rarely serious and for
which preventive or therapeutic
interventions are often available.
Campylobacter coli, Salmonella, P. falciparum,
Hepatitis viruses
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A pathogen that can cause human or
animal disease but is unlikely to be a
serious hazard to laboratory workers, the
community, livestock or the environment.
Laboratory exposures may cause serious
infection, but effective treatment and
preventive measures are available and the
risk of spread of infection is limited.
(Moderate individual risk; low community
risk)
Overview of Biosafety Levels (BSL) and Risk Groups (RG)
RISK GROUP
NIH GUIDELINES FOR RESEARCH
CLASSIFICATION INVOLVING RECOMBINANT DNA
MOLECULES 2002
WORLD HEALTH ORGANIZATION
LABORATORY BIOSAFETY MANUAL
3RD EDITION 2004
Risk Group 3
A pathogen that usually causes serious
human or animal disease but does not
ordinarily spread from one infected
individual to another. Effective treatment
and preventive measures are available.
(High individual risk; low community risk)
Agents that are associated with serious or
lethal human disease for which preventive
or therapeutic interventions may be
available
M. tuberculosis, R. tsutsugamushi, Japanese
encephalitis virus, HIV
Risk Group 4
Agents that are likely to cause serious or
lethal human disease for which preventive
or therapeutic interventions are not
usually available
Ebola virus, Tick-borne encephalitis virus
complex , Herpesvirus simiae (Herpes B or
Monkey B virus)
A pathogen that usually causes serious
human or animal disease and that can be
readily transmitted from one individual to
another, directly or indirectly. Effective
treatment and preventive measures are
not usually available.
(High individual and community risk)
available.
http://www.absa.org/riskgro
ups/index.html
Overview of Biosafety Levels (BSL) and Risk Groups (RG)
Biosafety Level
&
Risk Group
WHO 2004: Laboratory Biosafety Manual
Overview of Biosafety Levels (BSL) and Risk Groups (RG)
Overview of Biosafety Levels (BSL) and Risk Groups (RG)
CDC-NIH 2007: Biosafety in Microbiological and Biomedical Laboratories (BMBL)
Summary of Recommended Biosafety Levels for Infectious Agents
BSL
1
2
AGENTS
Not known to consistently
cause diseases in healthy
adults
• Agents associated with
human disease
• Routes of transmission
include percutaneous
injury, ingestion, mucous
membrane exposure
PRACTICES
PRIMARY BARRIERS
AND SAFETY
EQUIPMENT
FACILITIES
(SECONDARY
BARRIERS)
Standard
Microbiological
Practices
BSL-1 practice plus:
• Limited access
• Biohazard warning
signs
• “Sharps” precautions
• Biosafety manual
defining any needed
waste decontamination
or medical surveillance
policies
None required
Laboratory bench
and sink required
Primary barriers:
BSL-1 plus:
• Class I or II BSCs or
• Autoclave
other physical
available
containment devices
used for all
manipulations of
agents that cause
splashes or aerosols
of infectious materials
PPEs:
• Laboratory coats;
gloves;face protection
as needed
Overview of Biosafety Levels (BSL) and Risk Groups (RG)
CDC-NIH 2007: BMBL
Summary of Recommended Biosafety Levels for Infectious Agents
BSL
AGENTS
PRACTICES
3
• Indigenous or exotic
agents with potential for
aerosol transmission
• Disease may have
serious or lethal
consequences
BSL-2 practice plus:
• Controlled access
• Decontamination of
all waste
• Decontamination of
laboratory clothing
before laundering
• Baseline serum
PRIMARY
BARRIERS AND
SAFETY
EQUIPMENT
Primary barriers:
• Class I or II BSCs
or other physical
containment
devices used for all
open manipulation
of agents
PPEs:
• Protective
laboratory
clothing; gloves;
respiratory
protection as
needed
FACILITIES
(SECONDARY
BARRIERS)
BSL-2 plus:
• Physical
separation from
access corridors
• Self-closing,
double-door
access
• Exhaust air not
recirculated
• Negative airflow
into laboratory
Biosafty Level 2 Requirement
Summary of Recommended Biosafety Levels for Infectious Agents
BSL
4
AGENTS
PRACTICES
PRIMARY
BARRIERS AND
SAFETY
EQUIPMENT
• Dangerous/exotic agents BSL-3 practices plus:
Primary barriers:
which pose high risk of life • Clothing change
• All procedures
threatening disease
before entering
conducted in Class
• Aerosol-transmitted
• Shower on exit
III BSCs or Class I or
laboratory infections have • All material
II BSCs in
occurred; or related agents decontaminated on exit combination with
with unknown risk of
from facility
full-body, airtransmission
supplied, positive
pressure personnel
suit
FACILITIES
(SECONDARY
BARRIERS)
BSL-3 plus:
• Separate building
or isolated zone
• Dedicated supply
and exhaust,
vacuum, and
decontamination
systems
• Other
requirements
outlined in the text
Overview of Biosafety Levels (BSL) and Risk Groups (RG)
Biological risk assessments
• Provide a guide for the selection of appropriate biosafety
levels and microbiological practices, safety equipment, and
facility safeguards that can prevent LAIs
• Responsibility: Directors and principal investigators of,
Institutional biosafety committees (IBC), Animal care and use
committees, Biological safety professionals, and Laboratory
animal veterinarians
Overview of Biosafety Levels (BSL) and Risk Groups (RG)
Risk Criteria for Establishing Ascending Levels
of Containment
•
•
•
•
Safe
Practice
Infectivity
Safety
Severity of disease
Equipme
nt
Transmissibility
Nature of the work being
conducted
• Origin of the agent:
Indigenous or Exotic
Facility
Design
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Reduce/eliminate risk of infection
Exposure
1. Identify Hazards
2. Assess Hazards
Environmental and Procedural
5. Supervise
and
Evaluate
4. Implement
Controls
3. Develop
Controls
Make Decisions
BSL ?
Biosafty Level 2
Standard Microbiological Practices
Special Practices
Safety Equipment
Safe Practice
Laboratory Facilities
Safety
Equipment
Facility
Design
Biosafety Level 2: Standard Microbiological Practices
• Control access to the
laboratory.
•
Wash their hands after
working and before leaving
the laboratory.
•
Safely handle of sharps.
•
Perform all procedures to
minimize the creation of
splashes and/or aerosols.
Biosafety Level 2: Standard Microbiological Practices
•
Decontaminate work
surfaces after completion of
work and after any spill or
splash.
Disinfectant
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• Decontaminate all cultures,
stocks, and other potentially
infectious materials before
disposal.
•
Implement an effective
integrated pest management
program.
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2009/...hem.html
Biosafety Level 2: Standard Microbiological Practices
• Laboratory supervisor must
ensure that laboratory
personnel receive
appropriate training:
 Duties
 Necessary precautions to
prevent exposures
 Exposure evaluation
procedures.
• Personnel must receive
annual updates or additional
training.
Biosafety Level 2: Standard Microbiological Practices
•
Personal health status may
impact an individual’s
susceptibility to infection,
ability to receive
immunizations or
prophylactic interventions.
Biosafety Level 2: Standard Microbiological Practices
• Post a sign incorporating the
universal biohazard symbol
at the entrance to the
laboratory. Information must
include:
 Biosafety level
 Supervisor’s name (or other
responsible personnel)
 Telephone number,
 Required procedures for
entering and exiting the
laboratory
 Agent information
Biosafety Level 2: Standard Microbiological Practices
Do Not
• Eat, drink, smoke, handle
contact lenses, apply
cosmetics, and store food for
human consumption in
laboratory areas.
• Store food outside the
laboratory area.
• Mouth pipette.
Biosafety Level 2: . Special Practices
• Advised all persons entering
the laboratory of the
potential hazards and meet
specific entry/exit
requirements.
•
Prepare and adopt a
laboratory-specific biosafety
manual as policy. The manual
is available and accessible.
Biological Safety Manual Goals
1. Protect employees, the surrounding community and the
environment during activities involving potentially
hazardous biological agents.
2. Provide minimum technical safety and occupational
health requirements and guidelines for developing and
control process for operation involving etiologic agents.
3. Provide an environment for high quality research while
maintaining a safe work place.
4. Comply with applicable federal, state and local
requirements .
Biosafety Level 2: . Special Practices
•
Laboratory supervisor must
ensure that laboratory
personnel demonstrate
proficiency in standard and
special microbiological
practices before working.
AFRIMS Department of Enteric Disease Training 2005
Biosafety Level 2: . Special Practices
• Provided Laboratory
personnel medical
surveillance and offer
appropriate immunizations.
•
Store a baseline serum
sample, when appropriate.
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Biosafety Level 2: . Special Practices
• Place potentially infectious
materials in a durable, leak
proof container during
collection, handling,
processing, storage, or
transport within a facility.
• Decontaminate laboratory
equipment routinely.
• Decontaminate equipment
before repair, maintenance,
or removal from the
laboratory.
Biosafety Level 2: . Special Practices
• Contain, decontaminate, and
clean up spills by staff
properly trained.
Biosafety Level 2: . Special Practices
• Immediately evaluated and
treated incidents.
• Report all such incidents to
the laboratory supervisor.
• Provide medical evaluation,
surveillance, and treatment
and maintain appropriate
records.
Biosafety Level 2: . Special Practices
•
Conduct all procedures that
may generate an aerosol in a
BSC or other physical
containment devices.
• Do not permit animals and
plants not associated with
the work being performed in
the laboratory.
Biosafety Level 2: Safety Equipment (Primary Barriers
and Personal Protective Equipment)
• Use properly maintained
BSCs (preferably Class II)
• Appropriate PPEs
• Other physical containment.
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otinc.com
• Laminar Flow (LF):
- Product protection (no personnel protection)
- Not for biohazard agents or chemical fumes
• Fume Hood:
- Removes toxic chemical (ducting
sys./ductless)
- No HEPA filter -> not for biohazard agents
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• Biohazard Safety Cabinet (BSC)
- Product and personnel protection (not Class
1)
web.princeton.edu/.../labsafetymanual/bsc1.jpg
BSC Class
I
Airflow pattern
In at front
Exhausted through
HEPA to the outside*
or into the room.
Airflow pattern
Supply air inlets and
hard-duct exhausted
to outside through
two HEPA filters in
series.
III
Protection
Sample
Χ
Operator
√
Environment
√
Application with Chemicals
Nonvolatile Toxic Chem
Radionuclide
√
Volatile Toxic
Radionuclide
√*
Protection
Sample
√
Operator
√
Environment
√
Application with Chemicals
Nonvolatile Toxic Chem
Radionuclide
Volatile Toxic Chem
Radionuclide
BMBL 5th
√
√
(Small
amount)
BSC Class
Airflow pattern
70% recirculated to the
cabinet work area
through HEPA
II A1
Exhaust air recirculate
to the laboratory or
discharged from the
building via a canopy
connection.
Sample
√
Operator
√
Environment
√
Application with Chemicals
Nonvolatile Toxic Chem
Radionuclide
√
Volatile Toxic Chem
Radionuclide
Χ
Protection
II A2 (A/B3)
Airflow pattern
Same as II, A1
Exhaust air recirculate
to the laboratory or
discharged from the
building via a canopy
connection*.
BMBL 5th
Protection
Sample
√
Operator
√
Environment
√
Application with Chemicals
Nonvolatile Toxic Chem
Radionuclide
Volatile Toxic Chem
Radionuclide
√
√*
(minute
amounts
)
Biosafety Level 2: Safety Equipment (Primary Barriers
and Personal Protective Equipment)
• Remove protective clothing
before leaving for nonlaboratory
• Dispose of or deposit for
laundering protective
clothing appropriately.
• Persons who wear contact
lenses in laboratories should
wear eye protection.
Biosafety Level 2: Safety Equipment (Primary Barriers
and Personal Protective Equipment)
• Select glove base on an
appropriate risk assessment.
Gloves must not be worn
outside the laboratory.
 Remove gloves and wash hands
when work with hazardous
materials has been completed
and before leaving the
laboratory.
 Do not wash or reuse
disposable gloves.
 Dispose of used gloves with
other contaminated laboratory
waste.
Biosafety Level 2: Laboratory Facilities (Secondary
Barriers)
• Laboratory doors should be
self-closing and have locks.
•
Laboratories must have a
sink for hand washing.
•
The laboratory should be
designed so that it can be
easily cleaned and
decontaminated. Carpets
and rugs in laboratories are
not permitted.
Biosafety Level 2: Laboratory Facilities (Secondary
Barriers)
• Laboratory furniture must
be capable of supporting
anticipated loads and uses.
•
Bench tops must be
impervious to water and
resistant to heat, organic
solvents, acids, alkalis, and
other chemicals.
•
Chairs used in laboratory
work must be covered with a
non-porous material.
Biosafety Level 2: Laboratory Facilities (Secondary
Barriers)
• Laboratory windows that
open to the exterior are not
recommended. Otherwise,
must be fitted with screens.
•
Vacuum lines should be
protected with HEPA filters,
or their equivalent. Liquid
disinfectant traps may be
required.
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Biosafety Level 2: Laboratory Facilities (Secondary
Barriers)
• BSCs must be installed
so that fluctuations of
the room air supply and
exhaust do not interfere.
Biosafety Level 2: Laboratory Facilities (Secondary
Barriers)
• An eyewash station must be
readily available.
• Mechanical ventilation
systems provide an inward
flow of air without
recirculation to spaces
outside of the laboratory.
Biosafety Level 2: Laboratory Facilities (Secondary
Barriers)
•
HEPA filtered exhaust air
from a Class II BSC can be
safely re-circulated back into
the laboratory environment
if the cabinet is tested and
certified at least annually.
web.princeton.edu/.../labsafetymanual/bsc1.jpg
Biosafety Level 2: Laboratory Facilities (Secondary
Barriers)
• A method for
decontaminating all
laboratory wastes should be
available in the facility
(e.g., autoclave, chemical
disinfection, incineration, or
other validated
decontamination method).
Disinfectant
http://i.ehow.com/images/GlobalPhoto/Articles/4492263/clorox2-main_Full.jpg
Autoclave
What should not be autoclaved?
•
•
•
•
•
Corrosive chemicals
Flammable chemicals
Combustible
Explosive
Radioactive materials
http://www.phac-aspc.gc.ca/msds-ftss/index-eng.php#s
Animal Biosafty Level 2
Standard Microbiological Practices
Special Practices
Safety Equipment
Safe Practice
Laboratory Facilities
Safety
Equipment
Facility
Design
Animal Biosafety Level 2: Standard Microbiological Practices
• Prior to beginning a study
animal protocols must be
reviewed and approved by
the IACUC and the
Institutional Biosafety
Committee.
• A safety manual specific to
the animal facility is
prepared or adopted
• The need for an animal
allergy prevention program
should be considered.
Animal Biosafety Level 2: Laboratory Facilities
(Secondary Barriers)
• The animal facility is
separated from areas that
are open to unrestricted
personnel traffic within the
building.
• External facility doors are
self-closing and self-locking.
www.dearborn-animals.com/New%252...plan.htm
• Doors to areas where
infectious materials and/or
animals are housed, open
inward, are self-closing.
Animal Biosafety Level 2: Laboratory Facilities
(Secondary Barriers)
• Sink traps are filled with
water, and/or appropriate
liquid to prevent the
migration of vermin and
gases.
• Floor drains must be
maintained and filled with
water, and/or appropriate
disinfectant to prevent the
migration of vermin and
gases.
home.howstuffworks.com/home-impr...ains.htm
www.diychatroom.com/f7/do-we-hav.../index3/
Animal Biosafety Level 2: Laboratory Facilities
(Secondary Barriers)
• External windows are not
recommended; if present,
windows should be sealed and
must be resistant to breakage.
• Cages should be autoclaved or
otherwise decontaminated
prior to washing. Mechanical
cage washer should have a
final rinse temperature of at
least 180°F.
www.detach.se/products%2520-%252...hing.htm
References
• American Biosafety Association
http://www.absa.org/riskgroups/index.html.
• CDC-NIH, 2007. Biosafety in Microbiological and
Biomedical Laboratories (Fifth Edition).
• WHO, 2004. Laboratory Biosafety Manual.
Thank You and Questions?