Transcript MSSM Biosafety Training 1 CDC Slides Based on BMBL 4th Edition  Part  Part 2 MSSM specific training and over view of practices.

MSSM Biosafety Training
1 CDC Slides Based on BMBL 4th
Edition
 Part
 Part
2 MSSM specific training and over
view of practices
Biosafety Training
1 CDC Slides Based on BMBL 4th
Edition
 Part
 More
Biosafety Information is found at:
http://www.cdc.gov/od/ohs/default.htm
Biosafety in Microbiological
and Biomedical Laboratories
4th Edition
JY Richmond
&
RW McKinney (eds.)
BMBL
Introduction
 1941 - Meyer and Eddie
 74 lab associated brucellosis infections in
US
 1949 - Sulkin and Pike
 222 viral infections (21 fatal)
 Only 27% related to known accidents
BMBL
Introduction
 1951,




1965, 1976 - Sulkin and Pike
Surveys for lab-associated infections
More than 5,000 labs
Cumulative total of 3,921 cases cited
Most commonly reported:
• Hepatitis
• Tuberculosis
• Typhoid
• Venezuelan Equine Encephalitis
• Brucellosis
• Tularemia
BMBL
Introduction
 1951,1965, 1976 - Sulkin and Pike
 Fewer than 20% associated with known
accidents
 Exposure to infectious aerosols plausible (but
unconfirmed) for >80% of reported cases
Principles of Biosafety
Introduction
 Biosafety
Levels (BSLs)
 Laboratory Practice and Technique


Standard Practices
Special Practices
 Safety
Equipment (Primary Barriers)
 Facility Design and Construction
(Secondary Barriers)
Principles of Biosafety
Biosafety Levels
Biosafety Levels 1-3 provide:


Increasing levels of personnel and
environmental protection
Guidelines for working safely in
microbiological and biomedical laboratories
Principles of Biosafety
Laboratory Practice and Technique
Requirements:



Knowledgeable supervisor
Personnel
• Aware of potential hazards
• Proficient in practices & techniques
Biosafety manual specific to lab
Principles of Biosafety
Safety Equipment (Primary Barriers)
 Biosafety
cabinets (BSCs) - BSL 2/3
 Personal protective clothing


Gloves
Gowns
 Pipetting
Devices
 Safety centrifuge cups and rotors
 Eye and face protection
 Respiratory protection - BSL 3
Biosafety Level 1
Introduction
Suitable for work involving wellcharacterized agents not known to cause
disease in healthy adult humans and of
minimal potential hazard to laboratory
personnel and the environment.
Biosafety Level 1
Introduction
Examples:




Bacillus subtilis
Naegleria gruberi
Infectious canine hepatitis virus
E. coli
Biosafety Level 1
Facility Design (Secondary Barrier)
Biosafety Level 1
Facility Design (Secondary Barrier)
Requirements:






Laboratories have doors
Sink for hand washing
Work surfaces easily cleaned
Bench tops are impervious to water
Sturdy furniture
Windows fitted with flyscreens
Biosafety Level 1
Facility Design (Secondary Barrier)
Easily cleaned and
decontaminated
Biosafety Level 1
Facility Construction (Secondary Barrier)
Requirements:

Location - not separated
Structure - normal construction

Ventilation - none

Biosafety Level 1
Standard Microbiological Practices

Use mechanical pipetting devices

Wash hands

Restrict or limit access when working

Prohibit eating, drinking and smoking
Biosafety Level 1
Standard Microbiological Practices
Use
mechanical
pipetting
devices
Biosafety Level 1
Standard Microbiological Practices
Wash hands
Biosafety Level 1
Standard Microbiological Practices (cont.)

Minimize splashes and aerosols

Decontaminate work surfaces daily

Decontaminate wastes

Maintain insect & rodent control program
Biosafety Level 1
Safety Equipment (Primary Barriers)
Protective clothing


Lab coat
Gloves
Biosafety Level 1
Safety Equipment (Primary Barriers)
Personal protective equipment


Face protection
Eye protection
Biosafety Level 1
Special Practices
None required
Biosafety Level 1
Training Requirements
Supervisor

Scientist with general training in microbiology or
related science
Lab

Personnel
Specific training in lab procedures
Biosafety Level 2
Introduction
Suitable for work involving agents
of moderate potential hazard to personnel
and the environment.
Biosafety Level 2
Introduction
Examples:




Measles virus
Salmonellae
Toxoplasma spp.
Hepatitis B virus
* Immunization or antibiotic treatment is
available
Biosafety Level 2
Introduction
Examples:


Bloodborne pathogens
Human body fluids/particularly when visibly
contaminated with blood
* Extreme precaution with contaminated
needles or sharp instruments
Biosafety Level 2
Facility Design (Secondary Barriers)
Biosafety Level 2
Facility Design (Secondary Barriers)
Requirements:






Laboratories have lockable doors
Sink for hand washing
Work surfaces easily cleaned
Bench tops are impervious to water
Sturdy furniture
Biological safety cabinets installed as needed
Biosafety Level 2
Facility Design (Secondary Barriers)
Requirements (cont.):




Adequate illumination
Eyewash readily available
Air flows into lab without re-circulation to non-lab
areas
Windows fitted with flyscreens
Biosafety Level 2
Facility Design (Secondary Barrier)
Restricted access
when work in
progress
Biosafety Level 2
Facility Construction (Secondary Barrier)
Requirements:

Location - separated from public
areas

Structure - normal construction

Ventilation - directional
Biosafety Level 2
Standard Microbiological Practices

As in BSL-1
Biosafety Level 2
Special Practices
Needles & Sharps Precautions


Use sharps containers
DON’T break, bend, re-sheath or reuse
syringes or needles
Biosafety Level 2
Special Practices
Needles & Sharps Precautions (cont.)

DON’T place needles or sharps in office waste
containers
Biosafety Level 2
Special Practices
Needles and Sharps Precautions (cont.)

DON’T touch broken glass with hands
Biosafety Level 2
Special Practices
Needles and Sharps Precautions (cont.)

Use plasticware
Biosafety Level 2
Special Practices
Policies
and procedures for entry
Biohazard warning signs
Biosafety manual specific to lab
Training with annual updates
Biosafety Level 2
Special Practices
 Use
leak-proof transport containers
Biosafety Level 2
Special Practices
 Immunizations
 Baseline
serum samples
Biosafety Level 2
Special Practices
 Decontaminate
 Report
 No
work surfaces
spills and accidents
animals in laboratories
Biosafety Level 2
Safety Equipment (Primary Barriers)
In addition to BSL-1:

Use biosafety cabinets (class II) for
work with infectious agents involving:
• Aerosols and splashes
• Large volumes
• High concentrations
Biosafety Level 2
Safety Equipment (Primary Barriers)
Class
II Biosafety
Cabinet

Airflow
Biosafety Level 2
Safety Equipment (Primary Barriers)
Class

II Biosafety Cabinet
Equipment layout
Biosafety Level 2
Safety Equipment (Primary Barriers)
Class

II Biosafety Cabinet
Technique
Biosafety Level 2
Laboratory Facilities (Secondary Barriers)
BSL-1
Facilities PLUS:


Autoclave available
Eyewash station
available
Biosafety Level 2
Special Practices
Supervision

Supervisor is a competent scientist with increased
responsibilities
• Limits access if immunocompromised
• Restricts access to immunized

Lab Personnel


Aware of potential hazards
Proficient in practices/techniques
Biosafety Level 3
Introduction
Suitable
for work with infectious
agents which may cause serious or
potentially lethal disease as a result
of exposure by the inhalation route.
Biosafety Level 3
Introduction



Exposure potential to pathogens spread
by aerosol
Infection serious, possibly lethal
Examples:
• M. tuberculosis
• St. Louis encephalitis virus
• Coxiella burnetii
Biosafety Level 3
Laboratory Facilities (Secondary Barriers)
 BSL-1
and 2 Facilities PLUS:

Separate building or isolated zone

Double door entry

Directional inward airflow

Single-pass air
Facility Design
(Tertiary Barriers)

CDC’s MCL
Facility Design
(Tertiary Barriers)

Lab structure
 Lab ventilation
Biosafety Level 3
Laboratory Facilities (Secondary Barriers)
Biosafety Level 3
Laboratory Facilities (Secondary Barriers)
 BSL-1



and 2 Facilities PLUS (cont.):
Enclosures for aerosol generating
equipment
Room penetrations sealed
Walls, floors and ceilings are water
resistant for easy cleaning
Biosafety Level 3
Special Practices
BSL-2



Special Practices PLUS:
Work in certified BSC
Use bioaerosolcontaining equipment
Decontaminate spills
promptly
Biosafety Level 3
Laboratory Facilities ( Secondary Barriers)
 BSL-1

and 2 Facilities PLUS:
Vacuum lines protected
Biosafety Level 3
Standard Microbiological Practices
 As
in BSL-1 and -2
Biosafety Level 3
Safety Equipment (Primary Barriers)
BSL-1

and 2 Safety Equipment PLUS:
BSC Class
II or III to
manipulate
infectious
material
Biosafety Level 3
Safety Equipment (Primary Barriers)
BSL-1

and 2 Safety Equipment PLUS:
Respiratory protection
may be indicated
Biosafety Level 3
Special Practices
Supervision

Supervisor is a competent scientist experienced
working with agents
•
•
•
•
Establishes criteria for entry
Restricts access
Develops policies/procedures
Trains lab personnel
Biosafety Level 3
Special Practices

Lab Personnel





Strictly follow guidelines
Demonstrate proficiency
Receive appropriate training
Report incidents
Participate in medical surveillance
Principles of Biosafety
Summary
 BSL 1-3
 Standard Practices
 Special Practices
 Safety Equipment (Primary Barriers)
 Laboratory Facilities (Secondary Barriers)
 Building (Tertiary Barriers)
END of CDC Slides
BIOSAFETY TRAINING
 Part
2 MSSM specific training and over
view of practices
Biosafety Principles
Philip G. Hauck, MS, MSHS, CBSP, SM(NRM)
Biosafety Principles
 What
is Biosafety?
 History
 Principles
 Microorganisms
 Risks
 Regulatory Agencies
Biosafety Principles
 Primary
Containment
 Secondary Containment
 Biological Containment
 Health and Safety Practices
 Applying Biosafety Principles
 Emergency Procedures
Biosafety Principles

Biosafety has an old and a new beginning. The “old”
beginning was initiated with the first studies of
microorganisms, disease and pathology. Aseptic
technique, disinfectants and culturing methods and
rudimentary safety devices were developed during the
late 1800’s-early 1900’s.

The “new” beginning came in the 1970’s at the time
when researchers developed the abilities to “cut-and splice” DNA, and transfect microorganisms with these
novel recombinants, requiring the development of a new
biosafety on the framework of the old principles.
Biosafety Principles
Good Microbiological Technique - there is no
substitute; good practices are the foundation
Possess beforehand the knowledge, skill and
ability to work with specific organisms safely and
in a manner that does not present risk either to
self or to others
Handwashing after handling cultures or
infectious materials is mandatory!!
Biosafety Principles
Good “Housekeeping”- keeping work surfaces and
equipment clean and disinfected; safe storage of
infectious materials at all times
Decontamination of stocks, cultures and wastes by
autoclave before removal /disposal from the lab
Controlling aerosol - generating practices and splash
hazards by using Biosafety Cabinets and safety
devices
Needles, syringes, glass-items and other sharps are
handled with caution and disposed of accordingly in
the Biosystems Sharps Container
Biosafety Principles
Microorganisms of interest
-Bacteria
-Chlamydiae
-Rickettsiae
-Fungi
-Viruses
-Protozoans(Parasites)
Biosafety Principles
a HAZARD is a characteristic that is
inherent within a particular microbe that
allows it to cause disease or illness
RISK is the chance that an exposure or
circumstance will occur that will bring this
microbe and its inherent hazard into contact
with you as the recipient
Biosafety Principles
Biosafety:
Attempts to clarify the hazards and the risks of working
with specific microorganisms
Seeks to provide an operating framework in which
research can proceed safely with that microbe
A framework is used instead of rigid rules to provide
flexibility in dealing with different groups, and sometimes,
within individual genera of microorganisms which may
have different hazards than a wild-type or other strain
Biosafety Principles
Types of exposures:
 Blood*
 Aerosol/airborne*
 Food-borne
 Water-borne
 Vector-borne
*common laboratory exposures
Biosafety Principles
Blood exposures:
direct contact with blood
indirect contact with blood-contaminated items
droplets and splashes from spills/releases
aerosols formed by laboratory manipulations
Biosafety Principles
Aerosol exposures:
1-5 micron diameter mists, particles, dusts or
spores – these can be suspended in air around
a spill or process that creates aerosols
can remain suspended for long periods of time
> 30-45 minutes
can be inhaled deep into alveolar regions of the
lung and establish an infection if the organism
causes respiratory diseases
Biosafety Principles
Other Routes of Exposure:
Needle(sticks)
Animal / Human Bites
Ingestion (Mouth Pipetting)
Ingestion (Secondary Fomite
Contamination)
Biosafety Principles
The “Chain” of Infection
organism is capable of causing disease
host (susceptible) for that given organism
dose routinely capable of infecting 50%
of a population and causing disease
route of exposure known to cause disease
virulence, the ability to overwhelm host
defences (i.e. strength of organism)
Biosafety Principles

Any interdiction in the Chain of Infection, such
as:




Dose reduction through containment or PPE
Use of non-human pathogens / non-replicating strains
Vaccination (render host unsusceptible)
Interdict exposure route using BSC’s, safety
equipment
….Breaks the Chain of Infection and can minimize or
eliminate disease in lab workers
Biosafety Principles
Primary Containment: (Lab Practices)
good microbiological practices
self-sealing closures, screw caps on containers
Biological Safety Cabinets / Glove Boxes
lab coats, gloves, gowns, suits, respirators (PPE)
special safety caps or sealed centrifuge rotors
Biosafety Principles
Secondary Containment: (Facilities)
self-closing, self-sealing doors to rooms
double-door anterooms
nested labs (rooms within rooms) i.e. BSL-3’s in BSL-2’s
100% in /100% out, directional airflow systems with
zoned pressure differentials between rooms and corridors
isolated / contained plumbing and electrical services
Biosafety Principles
Biological Containment:
E. coli (K-12) Host-Vector systems
Bacillus subtilis Host-Vector Systems
Saccharomyces cerivisiae H-V systems
EK-1 Plasmid systems
EK-2 Plasmid systems
Biosafety Principles
Health and Safety Assesmment
Sources for Assistance:
ABSA
ASM
CDC
NIH
APIC
APHA
OSHA
http://absa.org/
http://www.asmusa.org/
http://www.cdc.gov/od/ohs/biosfty/biosfty.htm
http://www4.od.nih.gov/oba/rdna.htm
http://www.apic.org/
http://apha.org/
http://osha.gov
Public Health
http://www.phac-aspc.gc.ca/msds-ftss/index.html
Agency / Canada
Biosafety Principles
Applying Biosafety Principles
identify biological agents you will use
identify operations / manipulations used
identify possible releases, exposures encountered
consult the “BMBL” and “NIH Guidelines” to
identify the BSL needed for your protection for those
agents that have Agent Summary Statements
Biosafety Principles
Remember:
Source, Pathway, Receiver
Actions can be taken at any point in order to
intervene and reduce or eliminate a potential
exposure to a Biohazardous agent
Biosafety Principles
Source:
Pathway:
Receiver:
contain the microbes at the
point of release
Biosafety Principles
Source:
contain the microbes
Pathway: contain releases within
Class II or III cabinets; use
sealed centrifuge rotors;
plastic shields
Receiver:
Biosafety Principles
Source:
contain the microbes
Pathway:
contain releases within
Biosafety cabinets; sealed
centrifuge rotors
Receiver: contain YOU in protective
isolation, away from exposure in
Personal Protective Equipment,
such as respirators, gloves, waterimpervious garments
Biosafety Principles
Emergency Procedures
Have a:
Written Biosafety Manual or SOPs for hazardous work
Written Emergency procedures in case of exposure / spill
Contact numbers:
Biosafety Officer - 45169
Nurse- Call Page operator X41800 beep 4118 or X46023
Nights/wkends - Call Page operator X41800 and go to
ER!
ER - X47171
Security - ”60” and give information on the emergency
Biosafety Principles
References:
Chin, J; Control of Communicable Diseases Manual; 17th Edition 2000;
Am. Public Health Association, Washington, D.C.
Collins, C.H.; Laboratory-Acquired Infections; 2nd Edition, (1988)
Butterworths &Co. Ltd.London
U.S. D.H.H.S./P.H.S.; CDC-NIH Biosafety in Microbiological and
Biomedical Laboratories, May, 1999 ( 4th Ed.). Joint Publication by
NIH, Bethesda, MD and C.D.C. Atlanta Georgia. HHS No. (CDC)
93-8395
U.S. DHEW/P.H.S.: Guidelines for Research involving Recombinant DNA
Molecules; (January, 2001); National Institutes of Health.
U.S. D.H.H.S./P.H.S.;C.D.C. and NIH; Primary Containment for
Biohazards: Selection, Installation and Use of Biological Safety
Cabinets; 2nd Edition, Sept. 2000; CDC Atlanta, Ga.