BIOLOGICAL SAFETY IN

LABORATORIES
 BIOLOGICAL SAFETY IN LABORATORIES
LAMINAR FLOW CABINETS
LAMINAR OR UNI-DIRECTIONAL MEANS STRAIGHT UNINTERRUPTED
AIRFLOW
LAMINAR FLOW CABINETS ARE GENERALLY USED WHERE PROTECTION
IS REQUIRED FOR THE WORK OR PRODUCT ONLY
TWO TYPES ARE GENERALLY AVAILABLE
HORIZONTAL THE MAIN HEPA FILTER IS PLACED IMMEDIATELY
BEHIND THE WORKING AREA IN A VERTICAL POSITION, CLEAN AIR IS
BLOWN NORMALLY AT 0.45M/S ACROSS THE WORK SURFACE
TOWARDS THE OPERATOR.
VERTICAL THE MAIN HEPA FILTER IS PLACED IMMEDIATELY ABOVE
THE WORKING AREA, CLEAN AIR IS BLOWN NORMALLY AT 0.35M/S
VERTICALLY DOWN OVER THE WORK SURFACE.
BIOLOGICAL SAFETY IN LABORATORIES
BIOLOGICAL SAFETY CABINETS
THERE ARE THREE TYPES OF BIOLOGICAL SAFETY CABINETS

CHOOSING A CABINET SHOULD BE DONE TO ASSESS THE HAZARD

LEVELS OF MATERIAL BEING HANDLED IN CONJUNCTION WITH:-

A.C.D.P GUIDELINES ADVISORY COMMITTEE ON DANGEROUS

PATHOGENS

SAFETY OFFICER
BIOLOGICAL SAFETY IN LABORATORIES
CLASS 1
A CLASS 1 MICROBIOLOGICAL SAFETY CABINET HAS AN AIR FLOW
INTO THE CABINET IN ORDER TO PROTECT THE OPERATOR ONLY FROM
POTENTIAL AEROSOLS PENETRATING IN AN OUTWARD DIRECTION
FROM THE CABINET

A HEPA FILTER IS FITTED IMMEDIATELY ABOVE THE WORKING AREA

WHERE PARTICULATE IS RETAINED AS AIR PASSES THROUGH UNDER A
NEGATIVE PRESSURE

THE EXHAUST AIR IS EITHER RETURNED TO THE LAB IN A

RECIRCULATING MODE OR EXHAUSTED TO ATMOSPHERE
BIOLOGICAL SAFETY IN LABORATORIES
THIS CABINET WOULD BE USED TYPICALLY IN LABORATORIES WHERE
PATIENT SPECIMENS ARE HANDLED PRIOR TO IDENTIFICATION OF ANY
DISEASE

CLASS 1 CABINETS ARE GENERALLY EXHAUSTED TO ATMOSPHERE

MATERIALS IN HAZARD GROUP 2 AND 3 CATEGORIES ARE NORMALLY

USED IN CLASS 1 CABINETS, SUBJECT TO PROTECTION REQUIRED
BIOLOGICAL SAFETY IN LABORATORIES
Class I Biological Safety Cabinets
Provide operator and environmental protection but
gives no product protection.

Class I cabinets are

open-fronted safety
cabinets

The air is drawn from

the room into the
work opening and it
passes the operator and
the product before it
leaves the cabinet via a
HEPA filter. Normally
ducted to atmosphere.
BIOLOGICAL SAFETY IN LABORATORIES
CLASS 2

A CLASS 2 BIOLOGICAL SAFETY CABINET HAS TWO PROTECTION

FACTORS

PRIMARILY OPERATOR PROTECTION

SECONDARY PRODUCT LAMINAR DOWN FLOW PROTECTION

THIS CABINET WOULD BE USED TYPICALLY FOR TISSUE CULTURE, MEDIA

PREPARATION ETC WHERE A STERILE WORKING AREA IS REQUIRED
FOR PATIENT TRANSFER
BIOLOGICAL SAFETY IN LABORATORIES
Class II Safety Cabinets
Give both Operator and Work protection with Laminar downflow for
work and inward airflow for operator.
Must not be confused with simple
Laminar flow cabinets.
BS EN 12469 states they can be
installed with either:
Single exhaust filter, or
Direct duct connection to extract system.
UK ACDP regulations ask for double HEPA
exhaust filter (AEF) only when working
with material of Hazard Groups higher than 2.
(Do not use Group 4!)
BIOLOGICAL SAFETY IN LABORATORIES- CLASS 2
CABINET
 BIOLOGICAL SAFETY IN LABORATORIES
CLASS 3

A CLASS 3 BIOLOGICAL SAFETY CABINET UNLIKE THE CLASS 1 AND 2 IS

A FULLY SEALED SHELL FOR OPERATOR PROTECTION

FULL BARRIER PROTECTION IS MAINTAINED

MANIPULATION OF PRODUCT IS DONE BY USING GLOVED SLEEVED

PORTS

AIR IS DRAWN INTO THE CABINET VIA A HEPA FILTER AND EXITS VIA
A FURTHER HEPA
 BIOLOGICAL SAFETY IN LABORATORIES
CLASS 3 CABINETS ARE GENERALLY USED FOR THE HIGHEST HAZARD
CATEGORY OF PRODUCT AND PROTECTION UNDER THE ACDP
GUIDELINES.

A FURTHER CLASS 3 CABINET DERIVATIVE IS THE CLASS 3 1

A REMOVABLE NIGHT DOOR ALLOWS THE CLASS 1 MODE TO BE USED

CLASS 3 CABINETS ARE GENERALLY DUCTED TO ATMOSPHERE

BIOLOGICAL SAFETY IN LABORATORIES
CLASS III CABINETS
Fan
Anti blow-back
valve + alarm Pre-filter

Exhaust Inlet
HEPA HEPA
INSTALLATION OF BIOLOGICAL SAFETY CABINETS
BIOLOGICAL SAFETY IN LABORATORIES
SITING AND TESTING OF SAFETY CABINETS IS VERY IMPORTANT

BSEN 12469:2000 REPLACED BS5726

BS5726:2005 HAS BEEN RE-ISSUED TO ACCOMMODATE SITING

CLASS 1 AND 2 CABINETS MUST BE KI TESTED AS PART OF

COMMISSIONING PROTOCOL
 Recommendation for Cabinet Positioning
BIOLOGICAL SAFETY IN LABORATORIES
SEPARATION OF AN UNDISTURBED ZONE
AROUND A SAFETY CABINET FROM BSC
TRAFFIC ROUTES 1000

SPACING WHEN THE SAME OPERATOR

USES A SAFETY CABINET AND THE BENCH
Bench
TOP OPPOSITE, OR WHERE ONLY BSC
top

OCCASIONAL TRAFFIC IS ANTICIPATED 1500

SPACING DETERMINED BY AIRFLOW

REQUIREMENTS WITH AN OPPOSING
BSC
WALL
2000
SPACING DETERMINED BY AIRFLOW
REQUIREMENTS WHEN SAFETY CABINETS
ARE OPPOSITE EACH OTHER BSC BSC

3000
Recommendation for Cabinet Positioning
BIOLOGICAL SAFETY IN LABORATORIES
PACING DETERMINED BY AIRFLOW BSC

REQUIREMENTS WITH ADJACENT SIDE WALLS 300

SPACING THAT AVOID UNDUE DISTURBANCE TO

300
AIRFLOW. FACE OF COLUMN NOT IN FRONT OF
PLANE OF CABINET APERTURE BSC

SPACING TO AVOID UNDUE DISTURBANCE TO

AIRFLOW WHEN FACE OF COLUMN IS IN FRONT 300

OF PLANE OF CABINET APERTURE BSC

SPACING THAT AVOID UNDUE

DISTURBANCES TO AIRFLOW
IN RELATION TO DOOR OPENINGS 1000
BSC
BSC

1500
BIOLOGICAL SAFETY IN LABORATORIES
FUME CUPBOARDS

FUME CUPBOARDS ARE DESIGNED TO PROTECT THE OPERATOR

USUALLY FROM A GASEOUS CHEMICAL OR SOLVENT HAZARD

BIOLOGICAL PROTECTION !!!!!

BIOLOGICAL SAFETY IN LABORATORIES

DUCTED FUME CUPBOARDS

FILTRATION FUME CUPBOARDS

BIOLOGICAL SAFETY IN LABORATORIES
INSTALLATION OF FUME CUPBOARDS

SITING AND TESTING IS VERY IMPORTANT

BS7258 CONSISTS OF 4 SEPARATE PARTS FROM USAGE TO

INSTALLATION AND SITING
 BIOLOGICAL SAFETY IN LABORATORIES
SUMMARY OF BOTH BIOLOGICAL SAFETY CABINETS AND FUME CUPBOARDS

BEFORE CHOOSING A CABINET

ASSESS THE LEVEL OF RISK

DECIDE THE CONDITIONS YOU REQUIRE

CHEMICAL OR BIOLOGICAL HAZARD

PRODUCT PROTECTION FACTOR

 BIOLOGICAL SAFETY IN LABORATORIES
GLOSSARY OF TERMS IN GENERAL USE

PRE FILTER

HEPA FILTER

D.O.P TEST

K.I DISCUS TEST

ACDP
BIOLOGICAL SAFETY IN LABORATORIES
DEFINITIONS OF HAZARD GROUPS

GROUP 1

GROUP 2

GROUP 3

GROUP 4
BIOLOGICAL SAFETY IN LABORATORIES
PRINCIPLES OF BIOSAFETY - LAB. PRACTICES AND TECHNIQUES
THESE WILL VARY ACCORDING TO THE LEVEL OF HAZARD
CATEGORY BEING STUDIED, BUT AS A ROUTINE THE FOLLOWING
PRACTICES ARE GENERALLY RECOMMENDED FOR ALL
LABORATORIES
LAB. COATS TO BE WORN
LAB. DOORS TO BE CLOSED DURING WORK
NO EATING, DRINKING OR SMOKING
NO MOUTH PIPETTING
MINIMISE AEROSOL RISK
WASH HANDS BEFORE LEAVING LAB.
SAFE STORAGE OF WASTE MATERIAL
ETC.
BIOLOGICAL SAFETY IN LABORATORIES
ROUTES OF LABORATORY INFECTION
AEROSOL INHALATION
INGESTION
INJECTION BY SYRINGE
ABSORPTION THROUGH SKIN
ABSORPTION THROUGH CUTS/ ABRASIONS
SOURCES OF LABORATORY INFECTION
BIOLOGICAL SAFETY IN LABORATORIES
THESE CAN CONTAIN HAZARDOUS BACTERIA, VIRUSES, FUNGI OR
PARASITES AND ALL LAB. SAMPLES SHOULD BE TREATED AS
POTENTIALLY DANGEROUS IN ROUTINE CLINICAL LABS.
BLOOD SAMPLES
URINE SAMPLES
SPUTUM SAMPLES
TISSUE SAMPLES
FAECES SAMPLES
POND/ RIVER WATER SAMPLES

RESEARCH LABS. MUST BE AWARE OF THEIR WORK HAZARDS AND

RISK TO THE STAFF
 BIOLOGICAL SAFETY IN LABORATORIES
PRINCIPLES OF BIOSAFETY - LAB. FACILITY
THERE ARE BASICALLY 4 TYPES OF LABORATORY FACILITY
BASIC LABORATORY (NO RISK)
CONTAINMENT LABORATORY (MEDIUM RISK)
HIGH CONTAINMENT LAB. (SEVERE RISK)
BIOLOGICAL SAFETY IN LABORATORIES
CONTAINMENT LEVEL 1
INSTRUCTION IN LAB. PROCEDURES
EASY TO CLEAN
LAB. DOOR CLOSED DURING WORK
LAB. COATS TO BE WORN
NO EATING, DRINKING, SMOKING
NO MOUTH PIPETTING
MINIMISE AEROSOLS
WASH HANDS BEFORE LEAVING LAB.
SAFE STORAGE OF WASTE MATERIAL
REPORT ALL ACCIDENTS
 BIOLOGICAL SAFETY IN LABORATORIES
CONTAINMENT LEVEL 1
HAZARD GROUP 1 - P1
DOORS CLOSED DURING WORK
NORMAL LAB. PROCEDURES
BIOLOGICAL SAFETY IN LABORATORIES
CONTAINMENT LEVEL 2
KEY ISSUES - AS PER LEVEL 1 PLUS THE FOLLOWING
TRAINING IN HANDLING PATHOGENS
WORK SUPERVISION
ADEQUATE SPACE FOR WORKER (24M3)
ELBOW TAPS
AUTOCLAVE IN SAME BUILDING
SPECIAL LAB. COATS
BIOHAZARD CABINETS SHOULD BE USED
DISINFECTING AFTER USE
BIOLOGICAL SAFETY IN LABORATORIES
CONTAINMENT LEVEL 2
HAZARD GROUP 1 - 2 / P1 -P2
USE CLASS II SAFETY CABINET
USE AUTOCLAVE
PROTECT AGAINST AEROSOL GENERATION

Class II

Auto
BIOLOGICAL SAFETY IN LABORATORIES
CONTAINMENT LEVEL 3
KEY ISSUES - AS PER LEVEL 2 PLUS THE FOLLOWING
FULL TRAINING
HIGH WORK SUPERVISION
RESTRICTED ACCESS TO LAB.
BIOHAZARD LABELS
NEGATIVE PRESSURE ROOM
LABORATORY AUTOCLAVE
GLOVES MUST BE WORN
BIOHAZARD CABINETS MUST BE USED
LAB. SPECIFIC EQUIPMENT
 BIOLOGICAL SAFETY IN LABORATORIES
CONTAINMENT LEVEL 3
HAZARD GROUP P1 -P3
KEEP ROOMS AT NEGATIVE PRESSURE SO AIR FLOWS FROM OUTDOORS
TO INDOORS
USE CLASS II SAFETY CABINET

Entry Change room

( ve) ( ve)

Class II
Minus
pressure
Auto
BIOLOGICAL SAFETY IN LABORATORIES
CONTAINMENT LEVEL 4
KEY ISSUES - AS PER LEVEL 3 PLUS THE FOLLOWING
CODE OF PRACTICE/ SAFETY OFFICER
SPECIFIC TRAINING/ FULL SUPERVISION
LAB. MUST BE SEALABLE
SEPARATE BUILDING
NEGATIVE PRESSURE VIA DOUBLE HEPA
ENTRY VIA AIRLOCK
INTERLOCKED AIR INLET/ EXHAUST
COMPLETE CLOTHING CHANGE
DOUBLE ENDED AUTOCLAVE
ALL WASTE TO BE MADE SAVE
BIOLOGICAL SAFETY IN LABORATORIES
CONTAINMENT LEVEL 4
HAZARD GROUP 4, P1 -P4
KEEP ROOMS AT NEGATIVE PRESSURE
USE CLASS III SAFETY CABINET Optional
air inlet
INSTALL AIRLOCK AND SHOWER

Restricted Change room

entry Minus
pressure

Shower

Class III
BIOLOGICAL SAFETY IN LABORATORIES
Filter Technology and Airflow Principle

Pre-filter Fan Plenum HEPA Work Area

Filter
BIOLOGICAL SAFETY IN LABORATORIES
Filter Technology and Airflow Principle

What is a HEPA Filter?

HEPA stands for High Efficiency Particulate Air filter.
They are normally 99.997% efficient against a particle size of
0.3m. (Eurovent EN 13091:1999, EN1822-1 Class H14 or
higher.)
What is an ULPA filter?
ULPA stands for Ultra Low Penetration Air filter.
They are normally 99.999% efficient against a particle size of
0.3m. (Eurovent Class U 14 or higher.)
BIOLOLOGICAL SAFETY IN LABORATORIES
Filter Technology and Airflow Principle
Filter construction
Air flow

Filter Medium

Gasket
Filter Frame
BIOLOGICAL SAFETY IN LABORATORIES
Filter Technology and Airflow Principle
Laminar Air Flow (Unidirectional)
c
v

c
v

Filter Media

Laminar Sheet
 BIOLOGICAL SAFETY IN LABORATORIES
Filter Technology and Airflow Principle
Diagram of Particle Filtration

1. Small Particle Impaction 2

2. Electrostatic Attraction
3. Brownian Motion
(very small particles)
3
4. Large Particle Impaction
4
BIOLOGICAL SAFETY IN LABORATORIES
DECONTAMINATION

FULL DECONTAMINATION IS ONLY NECESSARY INFREQUENTLY

IT IS NORMALLY CARRIED OUT BY USING FORMALDEHYDE VAPOUR
TO ENSURE THE FILTERS AND THE WHOLE CABINET IS STERILISED
IT MUST BE CARRIED OUT PRIOR TO ANY SERVICING ON THE CABINET
OR FILTERS
FORMALDEHYDE IS DUE TO BE REPLACED IN EARLY 2009 UNDER NEW
COSHH DIRECTIVE AND WILL BE REPLACED WITH VAPOROUS
HYDROGEN PEROXIDE (VHP)
BASIC BIOSAFETY PRINCIPLES

EH&S ACADEMY
BRENDA J. WONG, UCSD BIOSAFETY OFFICER
OCTOBER 2009
 WHAT IS BIOSAFETY?

SAFETY FROM EXPOSURE TO

INFECTIOUS AGENTS

SMALLPOX
 WHAT IS BIOSAFETY?

SUNDAY, SEP. 20, 2009

DID THE PLAGUE KILL ILLINOIS SCIENTIST?
BY AP

(AP / CHICAGO) THE UNIVERSITY OF CHICAGO MEDICAL CENTER SAYS THE INFECTION THAT
KILLED A SCIENTIST MAY BE CONNECTED TO BACTERIA HE RESEARCHED THAT CAUSES THE PLAGUE.

THE UNIVERSITY SAID SATURDAY THAT ITS RESEARCHER STUDIED THE GENETICS OF HARMFUL
BACTERIA INCLUDING YERSINIA PESTIS, WHICH CAUSES THE ILLNESS. HE DIED SEPT. 13. HIS NAME
AND AGE HAVEN'T BEEN RELEASED

THE MEDICAL CENTER SAYS THE BACTERIA HE WORKED WITH WAS A WEAKENED STRAIN THAT ISN'T
KNOWN TO CAUSE ILLNESS IN HEALTHY ADULTS. THE STRAIN WAS APPROVED BY THE CENTERS
FOR DISEASE CONTROL AND PREVENTION FOR LABORATORY STUDIES.

AN AUTOPSY FOUND NO OBVIOUS CAUSE OF DEATH BUT DID FIND THE PRESENCE OF THE BACTERIA.
MORE TESTS ARE PLANNED. NO OTHER ILLNESSES HAVE BEEN REPORTED.
 BIOSAFETY IN VARIOUS DISCIPLINES
BIOSAFETY IS RELATED TO SEVERAL
FIELDS
ECOLOGY: REFERRING TO IMPORTED
LIFE FORMS NOT INDIGENOUS TO
THE REGION (REGGIE THE
ALLIGATOR)
AGRICULTURE: REDUCING THE RISK
OF ALIEN VIRAL OR TRANSGENIC
GENES, OR PRIONS SUCH AS
BSE/"MADCOW; REDUCING THE
RISK OF FOOD BACTERIAL
CONTAMINATION
MEDICINE: REFERRING TO
ORGANS OR TISSUES FROM
BIOLOGICAL ORIGIN, OR GENETIC
THERAPY PRODUCTS, VIRUS;
LEVELS OF LAB CONTAINMENT
PROTOCOLS BSL-1, 2, 3, 4 IN
RISING ORDER OF DANGER
CHEMISTRY: I.E., NITRATES IN
WATER, PCB LEVELS AFFECTING
FERTILITY
EXOBIOLOGY: I.E., NASA'S POLICY
FOR CONTAINING ALIEN
MICROBES THAT MAY EXIST ON
SPACE SAMPLES - SOMETIMES
CALLED "BIOSAFETY LEVEL 5"
 BIOSAFETY IN ACADEMIC RESEARCH
RESEARCH UNIVERSITIES:
PROMOTING SAFE
LABORATORY PRACTICES, AND
PROCEDURES; PROPER USE OF
CONTAINMENT EQUIPMENT
AND FACILITIES; PROVIDES
ADVICE ON LABORATORY
DESIGN AND RISK ASSESMENT
OF EXPERIMENTS INVOLVING
INFECTIOUS AGENTS, RDNA
IN-VITRO AND IN-VIVO.

Bottom Line: Risk & Containment

 BIOHAZARD SYMBOL
CHARLES BALDWIN AT
NATIONAL CANCER
INSTITUTE AT NIH.
SYMBOL TO BE
MEMORABLE BUT
MEANINGLESS SO IT
COULD BE LEARNED.
BLAZE ORANGE
MOST VISIBLE UNDER
HARSH CONDITIONS
 BIOSAFETY ISSUES

LABORATORY SAFETY
BLOODBORNE PATHOGENS (BBP)
RECOMBINANT DNA (RDNA)
BIOLOGICAL WASTE DISPOSAL
INFECTIOUS SUBSTANCE AND
DIAGNOSTIC SPECIMEN SHIPPING
 BIOSAFETY ISSUES (CONT.)

RESPIRATORY PROTECTION
BIOTERRORISM AND SELECT AGENTS
MOLD AND INDOOR AIR QUALITY
OCCUPATIONAL SAFETY AND HEALTH IN THE USE OF RESEARCH
ANIMALS
BIOHAZARDS USED IN ANIMAL MODELS
 BIOHAZARDOUS MATERIALS

VIRUSES
BACTERIA
FUNGI
CHLAMYDIAE/RICKETTSIAE
 BIOHAZARDOUS MATERIALS
TRANSGENIC PLANTS, ANIMALS AND INSECTS
TRANSGENIC INSECTS
 BIOHAZARDOUS MATERIALS

HUMAN AND PRIMATE CELLS, TISSUES, AND BODY FLUIDS

BRAIN TISSUE FROM DEMENTED PATIENTS
VIRAL VECTORS
REPLICATION DEFICIENT VIRUSES
 Biosafety Concepts
Biosafety In Microbiological
and Biomedical Laboratories
BMBL (acronym)

CDC/NIH Publication
Safety Guidelines
Regulations of Institution receives
NIH funding
Code of Practice and Gold
Standard in Industry anl
Gold Standard
HHS Publication No. (CDC) 93-8395
Clinical & Research Lab.
 THE NEW BMBL

EARLY PRINT EDITION.

EMPHASIS ON RISK
& CONTAINMENT
 Biosafety Concepts
The BMBL
The BMBL continues
to be published by
the
CDC and the NIH

5th edition is now at

the printers

http://www.cdc.gov/od/ohs/bio
sfty/bmbl5/bmbl5toc.htm
ARE THE NIH GUIDELINES OPTIONAL?

GUIDELINES DOES NOT MEAN

OPTIONAL
THEY ARE A TERM AND
CONDITION OF NIH FUNDING FOR
RECOMBINANT DNA RESEARCH.
From Kathryn Harris, NIH, OBA
 BIOSAFETY CONCEPTS FROM
THE BMBL
PRINCIPLES OF BIOSAFETY
PRACTICE AND PROCEDURES
STANDARD PRACTICES
SPECIAL PRACTICES & CONSIDERATIONS

SAFETY EQUIPMENT
FACILITY DESIGN AND CONSTRUCTION
INCREASING LEVELS OF PROTECTION
 PRINCIPLES OF
BIOSAFETY
BIOSAFETY LEVELS 1-4 (BSL)
INCREASING LEVELS OF EMPLOYEE AND ENVIRONMENTAL
PROTECTION
GUIDELINES FOR WORKING SAFELY IN RESEARCH &
MEDICAL LABORATORY FACILITIES

ANIMAL BIOSAFETY LEVELS 1- 4

(ABSL)
LABORATORY ANIMAL FACILITIES
ANIMAL MODELS THAT SUPPORT RESEARCH
 BIOSAFETY
CONCEPTS
THE BMBL
(1) STANDARD MICROBIOLOGICAL
PRACTICES
MOST IMPORTANT CONCEPT / STRICT
ADHERENCE
AWARE OF POTENTIAL HAZARD
TRAINED & PROFICIENT IN TECHNIQUES
SUPERVISORS RESPONSIBLE FOR:
APPROPRIATE LABORATORY FACILITIES
PERSONNEL & TRAINING
SPECIAL PRACTICES & PRECAUTIONS
 BIOSAFETY
ISSUES
THE BMBL
(2) SAFETY EQUIPMENT
PRIMARY CONTAINMENT BARRIER
MINIMIZE EXPOSURE TO HAZARD
PREVENT CONTACT / CONTAIN AEROSOLS
ENGINEERING CONTROLS/ EQUIPMENT
PERSONAL PROTECTIVE EQUIPMENT (PPE)
GLOVES, GOWNS, RESPIRATOR, FACE SHIELD,
BOOTIES
BIOLOGICAL SAFETY CABINETS
COVERED OR VENTILATED ANIMAL CAGE
 BIOSAFETY
CONCEPTS
THE BMBL
(3) FACILITY DESIGN AND
CONSTRUCTION
SECONDARY BARRIER/ ENGINEERING
CONTROLS
CONTRIBUTES TO WORKER
PROTECTION
PROTECTS OUTSIDE THE LABORATORY
ENVIRONMENT & NEIGHBORHOOD
EX. BUILDING & LAB DESIGN,
VENTILATION, AUTOCLAVES, CAGE
WASH FACILITIES, ETC.
LABORATORY DESIGN

Warehouse Type Lab

 DISCUSSION

WHAT ARE SOME OF

THE NEGATIVES AND
POSITIVES OF THIS
OPEN LAB CONCEPT?
 BIOSAFETY LEVEL-1
CONCEPTS OF BIOSAFETY
BIOSAFETY LEVEL-1 (BSL-1 OR ABSL-1)
WELL CHARACTERIZED AGENTS
AGENTS NOT KNOWN TO CAUSE DISEASE (IN
HEALTHY HUMAN ADULTS; NOW HEALTHY IMMUNOCOMPETENT
ADULTS)
PROPHYLACTIC TREATMENT AVAILABLE
OPEN BENCH PROCEDURES
ANIMALS IN OPEN CAGE SYSTEM OR OPEN
ENVIRONMENT (OUTDOORS)
GOOD LABORATORY PRACTICES
 RISK GROUP 1 AGENTS

E.COLI K-12
TRANSGENIC PLANTS
 BSL-1 PRACTICES

BENCH-TOP WORK ALLOWED

DAILY DECONTAMINATION
MANUAL PIPETTING
REQUIRED HANDWASHING
RED BAG WASTE
BIO CABINET NOT REQUIRED
(UNLESS CREATING AEROSOLS)
2 CONTAINMENT
 RISK GROUP 2 AGENTS
HUMAN OR PRIMATE
CELLS
HERPES SIMPLEX
VIRUS
REPLICATION
INCOMPETENT
ATTENUATED HUMAN
IMMUNODEFICIENCY
VIRUS
PATIENT SPECIMENS
 BSL-2 PRACTICES
CONCEPTS OF BIOSAFETY
PRACTICES & PROCEDURES
AGENTS ASSOCIATED W/ HUMAN DISEASE
TREATMENT FOR DISEASE AVAILABLE
AGENT POSES MODERATE HAZARD TO
PERSONNEL AND ENVIRONMENT
DIRECT CONTACT OR EXPOSURE
PERCUTANEOUS EXPOSURE
SCRATCH, PUNCTURE, NEEDLE STICK
MUCUS MEMBRANE EXPOSURE
 BSL-2 PRACTICES

LIMITED ACCESS TO LAB

WHEN WORK IN PROGRESS
DAILY DECONTAMINATION
MECHANICAL PIPETTING
LABCOAT, SAFETY GLASSES
AND GLOVES REQUIRED
RED BAG & SHARPS
CONTAINERS REQUIRED
 BSL-2 PRACTICES (CONT)

BIOHAZ. SIGN POSTED AT

ENTRANCE TO LAB
LABEL ALL EQUIPMENT
(INCUBATORS, FREEZERS, ETC.)
TC ROOM NEGATIVE AIR FLOW
DOCUMENTED TRAINING
BASELINE SEROLOGY OR PRE-
VACCINATION MAY BE REQUIRED
 RISK GROUP 3 AGENTS

HUMAN
IMMUNODEFICIENCY
VIRUS
MYCOBACTERIUM
TUBERCULOSIS
COXIELLA BURNETII
 BIOSAFETY LEVEL 3
WORKING IN HIGH
CONTAINMENT

BIOSAFETY LEVEL-3 (BSL-3 OR

ABSL-3)
INDIGENOUS OR EXOTIC AGENTS
AEROSOL TRANSMISSION
SERIOUS HEALTH EFFECTS
TREATMENT MAY OR MAY NOT EXIST
 BSL-3 PRACTICES
PUBLIC ACCESS NOT PERMITTED
DAILY DECONTAMINATION AFTER SPILL AND
UPON COMPLETION OF EXPERIMENT
AUTOCLAVE REQUIRED AND WASTE IS
DISPOSED AT THE END OF DAY
REQUIRED FOOT ACTIVATED HANDWASHING
SINK AND CONTROLS
NO SHARPS UNLESS ABSOLUTELY NECESSARY
 BSL-3 PRACTICES (CONT)

AEROSOL MINIMIZATION PROCEDURES REQUIRED

WRAP AROUND DISPOSABLE CLOTHING IS REQUIRED.
SPECIALIZED EQUIPMENT MAY BE REQUIRED DEPENDING UPON
PROCEDURES
 BSL-3 PRACTICES (CONT)

BENCH TOP WORK NOT PERMITTED

DOCUMENTED TRAINING AND PERSONNEL
COMPETENCY CERTIFICATION (FOR BSL-3
PROCEDURES)
UCSDS BSL-3
 BIOSAFETY LEVEL-4
WORKING IN HIGH CONTAINMENT

BIOSAFETY LEVEL-4
BUILDS ON BSL-3/ ABSL-3 PRACTICES
MAXIMUM CONTAINMENT FACILITIES
PRESSURIZED CONTAINMENT SUITE
 BIOSAFETY LEVEL 4

LASSA FEVER VIRUS

EBOLA
HEMMORRHAGIC
FEVER VIRUS
MARBURG VIRUS
HERPES B VIRUS
 BIOSAFETY CONCEPTS
WORKING IN HIGH
CONTAINMENT

BIOSAFETY LEVEL-4 (BSL-4 OR ABSL-4)

DANGEROUS/EXOTIC AGENTS
LIFE THREATENING DISEASE
AEROSOL TRANSMISSION
AGENTS OF UNKNOWN RISK
OF TRANSMISSION OR HEALTH AFFECTS
NO KNOWN TREATMENT
ANIMAL BIOSAFETY LEVEL-4
WORKING IN HIGH CONTAINMENT
 GENERAL GOOD LAB TECHNIQUE

HYGIENIC PRACTICES
NO SMOKING, EATING, APPLYING COSMETICS, LIP BALM, CONTACTS
 GENERAL OPERATIONAL PRACTICES
PROPER ATTIRE
MINIMUM LAB COAT, SAFETY GLASSES, GLOVES

PLAN YOUR WORK

KNOW IN ADVANCE WHAT YOU ARE WORKING WITH
READ AVAILABLE RESOURCES (MSDS)
ANIMAL CONTAINMENT POINTS

CDC - 1957
CDC & UCSD - 2005

Courtesy of Paul Vinson, CDC

 DISCUSSION # 2

BASED ON WHAT YOU KNOW ABOUT BIOSAFETY LEVELS,

PRACTICES AND OPERATIONAL CONTROLS,
WHAT ARE SOME DISCUSSION ISSUES FOR
CONDUCTING BIOHAZARD RISK ASSESSMENTS?
HOW DO YOU APPROACH RISKS WHEN ADDRESSING A PARTICULAR
 RISK ASSESSMENT

In-Vitro In-Vivo Human Clinical

Trial
 ADDRESSING RISK ASSESSMENTS

WHAT IS THE ORGANISM?

IS IT WILD-TYPE, ATTENUATED, IRRADIATED,
OR CHEMICALLY TREATED?
LOOK AT KILL DATA OR KILL CURVES.
WHAT IS THE
 RISK ASSESSMENT, CONT
ARE PERSONNEL TRAINED? DO
PERSONNEL UNDERSTAND THE
ORGANISM, INFECTIOUS DOSE
AND SYMPTOMS?
WHAT ARE THEIR EXPERIMENTAL Tom Pugh

PROCEDURES?
WILL THEY BE TRANSPORTING THE
MATERIAL? SHIPPING INTRA, INTER-
STATE OR INTERNATIONAL?
ARE THEY DOING TISSUE CULTURE?
DO THEY HAVE ADEQUATE
CONTAINMENT EQUIPMENT?
 RISK ASSESSMENT, CONT
ARE THEY DOING THIS
WORK IN-VIVO? HAVE
YOU CONSULTED AND
DISCUSSED THIS WITH
THE VETS AND IACUC TO
DETERMINE SPECIAL
NEEDS AND HOUSING?
WASTE ISSUES
ADDRESSED?
PREGNANCY ISSUES
WITH THE ORGANISMS?
 RISK ASSESSMENT, CONT

DO THEY SHARE THEIR

TISSUE CULTURE ROOM?
DO THEY HAVE MORE THAN
1 BIOSAFETY CABINET?
OCCUPATIONAL HEALTH
INFORMED AND SET UP TO
RECEIVE PATIENT OR OFFER
COUNSELING?
 ACCIDENTAL SPILLS

Evacuate area, alert personnel and

cordon off so that aerosols may settle
Don PPE; Cover with paper towels
and apply bleach (1 part bleach : 9
parts water
Allow 15 20 min contact time
Wipe up working towards center
Use tongs if broken glass is involved
 FIRST AID MEASURES

SPLASH TO EYE OR NEEDLESTICK INJURY

RINSE THOROUGHLY FOR 15 MINUTES AT THE
EYEWASH OR SINK
CALL OCCUPATIONAL MEDICINE 619 471-9210
CALL EH&S TO REPORT EXPOSURE 858 534-5366
WHAT HELPS?