biosafety laboratories: containment vs exclusion · biosafety level 1 laboratory (bsl-1) • basic...

Biosafety Laboratories: Containment vs Exclusion Planning & Design Considerations

Janet Baum, AIA

Washington University in St. Louis

Containment Laboratories

• Maintain under negative pressure to ALL surroundings

• Treat effluent air •  Biosafety Labs •  High Toxicity Labs

Exclusion Laboratories

• Maintain under positive pressure to ALL surroundings

• Treat supply air •  Clean Room Laboratories •  Trace Analysis Laboratories

Neutral-Pressure Laboratories

• NO hazardous or toxic materials handled or stored

• NO volatile or malodorous materials handled or stored

• NO animals housed

Containment Laboratories

BIOSAFETY LABORATORY

•  Is a containment laboratory

• U.S.A. Biosafety Levels are from 1 to 4

• U.S.A. CDC / NIH designate hazard level of individual organisms on a scale of 1 to 4

• U.S.A. Dept. of Agriculture designates hazard level of organisms on a scale of 1 to 3

Centers for Disease Control with National Institutes of Health Biosafety in Microbiological and Biomedical Laboratories

U.S. Department of Agriculture

Agricultural Research Service (ARS) Facility Design Standards

Animal & Plant Health Inspection Service (APHIS) Quarantine Facility Guidelines for Microorganisms Quarantine Facility Guidelines for Weeds Quarantine Facility Guidelines for the Receipt and Containment Non-indigenous Arthropod Herbivores, Parasitoids, Predators

American Committee of Medical Entomology Arthropod Containment Guidelines

U.S.A. Standards and Guidelines

Biological Safety

National Institutes of Health Guidelines for Recombinant DNA Research Laboratory Safety Supplement to the Guidelines

Occupational Safety and Health Administration Bloodborne Pathogens Tuberculosis Standard

National Cancer Institute with NIH Guidelines for Oncogenic Viruses


Biological Safety

• World Health Organization Laboratory Safety Guidelines

• CEN (Comité Européen de Normalisation) CWA #15793 document, February, 2008, Laboratory Biorisk Management Standard

• International Union Against Tuberculosis and Lung Disease

1998 The Public Health Service National Tuberculosis Reference Laboratory Minimum Requirements, Role and Operation in a Low Income Country

International Standards & Guidelines

Biological Safety

AAALAC – Guide for Care and Use of Laboratory Animals Local Building Code National Building Code NFPA Standards Fire Protection Code Mechanical Code Plumbing Code Electrical Code Energy Code Nuclear Regulatory Commission OSHA Americans with Disabilities Act of 1990


Impacting Biosafety Laboratories

Public Law 107- 56: U.S.A. Patriot Act of 2001 (Uniting and Strengthening America by Providing Appropriate Tools Required to Intercept and Obstruct Terrorism)

Prohibits restricted persons from shipping, possessing, receiving select agents

Public Law 107- 188: U.S.A. Public Health Security and Bioterrorism Preparedness and Response Act of 2002 U.S.A. Agricultural Bioterrorism Protection Act of 2002

Requires institutions to notify DHHS or USDA of possession of specific pathogens or toxins

42 CFR Part 73 / Part 1003: U.S.A. Select Agent Act of 2002 (Possession, Use and Transfer of Select Agents and Toxins)

(Department of Health and Human Services)

U.S. Federal Laws

U.S.A. Regulations

U.S.A. Public Health Security and Bioterrorism Preparedness and Response Act of 2002 Requires DHHS to regulate the possession, use, and transfer of biological agents or toxins that could pose a severe threat to public health and safety. DHHS require entities to register with the U.S. Centers for Disease Control

U.S.A. Agricultural Bioterrorism Protection Act of 2002 Requires USDA to regulate the possession, use, and transfer of biological agents or toxins that could pose a severe threat to animal or plant health or to animal or plant products. USDA require entities to register with APHIS.

These Acts require establishment of a national database of registered entities Set criminal penalties for failing to comply with requirements of these Acts. CDC and APHIS coordinate regulatory activities for agents that would be Registered by both Agencies. The F.B.I. has been delegated authority for conducting security risk assessments.

U.S.A. Regulations Select Agents

U.S.A. Regulations

Viruses Bacteria Fungi & Rickettsae Toxins •  Hendra Virus -Equine

Morbillivirus •  Herpesvirus Simiae (B) •  HPAI Influenza •  1918 Influenza

• Arboviruse & Related Zoonotic Virus

•  Venezuelan Equine Encephalitis

•  Eastern Equine Encephalitis

•  Rift Valley Fever

•  Bacillus Anthracis •  Burcella Abortus •  Burkholderia Mallei •  Burkholderia Pseudomallei •  Neurotoxin Clostridium

(Botulinum) •  Francisella Tularensis

•  Coccidioides Immitis •  Coxiella Burnetii •  Rickettsia Prowazekii •  Rickettsia Rickettsii

•  Abrin •  Aflatoxins •  Botulinum •  Clostridium Perfringens Epsilon •  Conotoxin •  Diacetoxyscirpenol •  Ricin •  Saxitoxin •  Sigatoxin •  Staphylococcal Enterotoxin •  Tetrodotoxin •  T-2 Mycotoxin

CDC List of Select Agents

U.S.A. Regulations CDC List of Bacterial Agents

U.S.A. Regulations CDC List of Viral Agents

U.S.A. Regulations CDC List of Other Agents

Biosafety Level 1 Laboratory (BSL-1)

• Basic lab • Open bench operations • Standard lab practices • Operable windows have insect screens • Biosafety Cabinets (BSCs) not required

•  Biosafety cabinets should be used in introductory microbiology courses to teach students how to use BSCs.


in addition to all requirements for BSL-1

• Basic lab • Limited access into lab • Biohazard warning signs • Lab coats and protective gloves used • Decontamination facility needed in building • Biosafety Cabinets are required when operations may produce aerosols

• Negative pressure is achievable

Lab Planning Issues – BSL-2

Agents Practices Safety Equipment (Primary Barriers)

Facilities (Secondary Barriers)

Suitable for work involving agents of moderate potential hazard to personnel and the environment

BSL-1 plus:

• Controlled access

• Hand washing

• Keep lab clothing within lab, do not wear outside lab

• Baseline serum levels of staff

• Needles / sharps precautions

• Class II BSC’s or other physical containment devices used for all open manipulations of agents

• PPE: protective gear as needed

• Eyewash available

• Sink for hand wash

• Autoclave available

BSL-1 plus:

• Physical separation, & locked, self-closing doors from access corridors

• Sturdy, easily cleaned furniture and work surfaces

• Exhausted air not recirculated

• Negative airflow into laboratory

• Insect screens on windows

Goals of a Containment Facility

•  Protect personnel in the laboratory

•  Provide productive scientific environment –  Protect research, samples, and products

•  Protect support staff who operate and maintain containment and ventilation systems/ controls

•  Protect environment –  Surrounding personnel and functions –  Outside environment, adjacent communities

Liquid decon pass-thru

UV decon pass-thru pass-thru autoclave HEPA filter bank

Definition of a BSL-3 Facility

Suitable for work with infectious agents which may cause serious or potentially lethal disease as a result of exposure by the inhalation route •  Exposure potential to pathogens spread by aerosol •  Infection serious, possibly lethal

Canada National Biosafety Laboratory


in addition to all requirements for BSL-1 and BSL-2

• Designed specifically for biological containment

• Special protective clothing worn • All work with biological materials is in

biosafety cabinets (BSCs) • Access is through two doors separated by an

airlock that may be used for clothing change.

Lab Planning Issues – BSL-3

Agents Practices Safety Equipment (Primary Barriers)

Facilities (Secondary Barriers)

Indigenous or exotic agents with potential for aerosol transmission; disease may have serious or lethal consequences

BSL-2 plus:

• Controlled access

• Work in certified BSC

• Decontamination of all waste

• Decontaminate lab clothing before laundering

• Baseline serum levels of staff

• Class II or III BSC’s or other physical containment devices used for all open manipulations of agents

• PPE: protective lab clothing, gloves, respiratory protection as needed

• Autoclave in lab suite

• HEPA filter vacuum lines

BSL-2 plus:

• Physical separation from access corridors

• Locked self-closing, double-door access

• Exhausted air not recirculated

• Negative airflow into laboratory

• Room penetrations are sealed

• Room surfaces are water resistant for cleaning

Biosafety Level 3 Lab – ABSL-3

in addition to all requirements for BSL-1, BSL-2, BSL-3

• Designed specifically for biological containment of small research animals

• Entrance is through a pressure-controlled airlock used for clothing change.

• Exit optional through an airlock used for personnel showers and clothing change.

• Supply and exhaust systems separate and independent of other building ventilation.

•  Interlocked controls to maintain pre-set pressures

Biosafety Level 3 Lab – BSL-3Ag


• Designed specifically for biological containment of plants and large animals


• Exit is through an airlock used for personnel showers and clothing change.



Biosafety Level 4 Laboratory (BSL-4) in addition to all requirements for BSL-1, BSL-2, BSL-3

• Designed specifically for biological containment.


• Exit is through an airlock used for personnel showers and clothing change.

• Total containment Class III biosafety cabinets or

• Partial containment Class II biosafety cabinets used by personnel clothed in positive-pressure, full-body, suits with separate and protected air supply





• HEPA filtered supply air • Dual HEPA filtered exhaust air, in sequence •  In-place filter testing facilities

Cost Premium-Containment Facilities

Cost Premium-Containment Facilities

Diagram of BSL-4 Containment Concept

Containment Planning – BSL-3 and 4 Breathing

Air

Air Supply Diffuser

Waste Management All waste rendered harmless   autoclave waste from lab   decontaminate effluent waste   HEPA filter air

Camera Bubble

Floor/Sink Drains

Window Frame

Primary/Secondary Barriers Biosafety cabinets used

Gas tight room tested and certified

Air Locks with air pressure doors

Electrical Outlets

Containment Planning – BSL-3 and 4 •  Design of containment barriers •  Quality research = containment •  Ventilation requirements

BSL-3 lab in Canada Sealed pipe penetrations

Containment Planning – BSL-3 and 4

Containment Barriers

•  Primary barrier

–  Biological safety cabinet –  Personal protective gear

•  Secondary barrier –  Room enclosure –  Engineering systems

•  Tertiary barrier

–  Building around lab –  Containment around

systems

BSL-3 lab in Canada

Concentrate/ Minimize – BSL-3 and 4

•  Limit number of individuals and processes that must work under BSL-3 and BSL-4 conditions

•  Provide appropriate service support outside containment zone

•  Provide adequate functional support inside containment zone

•  Estimate capital and life cycle costs for operating facility

Capital cost includes design, construction, commissioning, and equipment fit-out of the laboratory

Life cycle cost includes utility expense for ventilation and electric power, filter testing / replacement, inspections, insurance / liability

Plan for Flexibility

• Regulatory and research changes • New pathogens • New equipment

• Hazard zoning • Egress in direction of lower hazard • Zone from clean to dirty • Separate hazards that may interact •  Isolate highest hazard processes & agents

Planning Concepts – BSL-3 and 4

Prep Lab Exit

Highest Hazard Lowest Hazard

Air Flow Contaminated

Hot Zone Hazard

Enter

•  One-way passage •  Enter BSL-3 lab from one passage, exit from

another •  Sequential work flow, if possible •  Reduce intersections in circulation pathways

Process Path

ENTRY

EXIT

Process #1

Process #5

Process #4

Process #3

Process #2


•  Suites

•  Cluster support functions around basic work zone to reduce distances between functional zones

•  Good for repetitive functions and processes


BSL-3

Culture Labs

Entry Vestibule

Specimen Drop-off

Decon

Equipment

Solutions/ Media Prep

BSL-3

Decon

Degown

AL BSL-2

AL

Gown

Cell Culture

BSL-3

Decon

Degown / Shower

AL

BSL-2 AL

Gown

BSL-3 Cell Culture

Micro Test AL

Graphic Program Blocking Diagram

Planning Concepts – BSL-3

BSL-3

Decon

Degown

AL

BSL-2 AL

Gown

BSL-3 Cell Culture

Micro Test AL

Flow Diagram

Personnel Supplies

Infectious Agent / Waste Material Animals


• Locate biosafety cabinets and fume hoods out of high traffic zones

• Develop understanding on how to deal with emergencies within containment zone

• Understand how to manage emergencies from outside zone

•  Investigate how containment zone can expand in future, if needed



Decon

Air Locks

Air Locks

Cell Culture

Micro Test AL

BS

C

FH

BS

C

BS

C

BS

C BS

C

BS

C

BS

C

AC

AC

Primary Entry and Exit

Emergency Exit Only

Shower

BSL-3 BSL-2

Degown & Gown

BS

C

Bench

EQ

EQ EQ

EQ

EQ EQ

Bench

Phones Phones


AL

Air Lock

Cell Culture AL Cell

Culture Decon Micro Test

Micro Test Decon

Air Lock

AL

BS

C

FH

BS

C

BS

C

BS

C

AC

AC

Shower

BSL-3 BSL-2

BS

C

Bench

EQ

EQ EQ

EQ

EQ EQ

Bench or

Equip

Phones Phones

Shower

BS

C

AC

AC

BSL-3

Bench or Equip

BSC BSC

BSC BSC

BS

C

EQ

Phones

Degown & Gown

Degown & Gown


Emergency Exit Only

Emergency Exit Only

Expansion Options


Expansion Options

Equip Decon

Cell Culture

BSL-3

Phones

BSL-2

Phones


Emergency Exit Only

Decon

Air Lock

AL

Micro Test AL

BS

C

FH

BS

C

BS

C

BS

C

AC

AC

Shower

BSC

Bench

EQ EQ

EQ

EQ EQ

Bench Equip

Or BSC

EQ

BSC

BSC

BSC BSC

BSC

EQ EQ

EQ

EQ

EQ

EQ Degown & Gown

• Understand how laboratory can be cleaned – Shape of each space is simple (minimize corners)

– Finishes are seamless, smooth and coved

– Seal ALL penetrations through interior surfaces, walls, floors, and ceilings

– Conduct Integrated Pest Management Program

– Equipment is located so floor can be cleaned; provide clearances above, below and beside them enough for mop and head of shop-vacuum machine

– Provide cleaning tools and supplies within lab for daily as well as emergency cleaning procedures

Design Concepts – BSL-3 and 4

• Understand how laboratory and equipment can be maintained – Provide tools for simple repairs

– Locate laboratory HVAC controls exterior to laboratory so maintenance staff do not have to enter containment zone to do tests and repairs

– Locate HVAC dampers and actuators exterior to laboratory so maintenance staff do not enter

– Clearances above, below and beside adequate for access to controls motors, fans, filters.


• Understand overall health & safety issues – Perform periodic risk assessments – Personnel Medical Surveillance Program – Ergonomics – Visibility to all parts of lab – Communications – internal and external – Alarm systems – internal and external – Safety equipment located in critical areas – Back-up systems for PPE, critical supplies


•  In developing countries 3 types of labs supported by W.H.O. and other NGOs

– Peripheral Laboratories in rural regions, villages for initial TB screening from sputum samples

– Intermediate Laboratories in towns and cities for initial TB screening and confirmation of

samples brought from Peripheral Laboratories

– Central Laboratories capitol of nation or region for initial TB screening, confirmation of samples

brought from Intermediate Laboratories, and testing for MDR and XDR TB

Design Concepts – TB Testing Labs

Design Concepts – WHO Level 3 PERIPHERAL LABORATORY 1


STORAGE CABINETS

REF

MICROSCOPE & WORK TABLES

WET BENCH LOG-IN

TABLE

HAND WASH SINK


LAB ENTRY

STORAGE CABINETS

REF MICROSCOPE & WORK TABLES &

STORAGE CABINET

WET BENCH

LOG-IN TABLE

HAND WASH SINK

WORK TABLE & STORAGE CABINET

Design Concepts – WHO Level 3 INTERMEDIATE LABORATORY 1


PPE ANTEROOM

SPUTUM RECEIVING & LOG-IN


PPE ANTEROOM

SPUTUM RECEIVING & LOG-IN

LAB ENTRY

MAIN ENTRY PASS-THRU

BOX

PASS-THRU BOX

LOG-IN TABLE

STORAGE CABINET

HAND WASH SINK


CULTURE LAB

INCUBATION LAB


CULTURE LAB

INCUBATION LAB

STORAGE SHELVES

BSC

HAND WASH SINK

CENTRI -FUGE

STORAGE SHELVES REF

MICROSCOPE TABLES

WET BENCH

PASS- THRU

BOX & TABLE

LAB ENTRY LAB

ENTRY


STERILIZATION LAB

MEDIA & REAGENT PREP LAB


STERILIZATION LAB

MEDIA & REAGENT PREP LAB

HAND WASH SINK

PASS-THRU AUTOCLAVE

WET BENCH PASS-

THRU BOX & TABLE

WET BENCH

PASS-THRU AUTOCLAVE

STORAGE CABINET

STORAGE CABINET

REF

LAB ENTRY


Mechanical Ventilation Zone 51 SM or 550 SF


Mechanical Ventilation Zone 51 SM or 550 SF

Natural Ventilation Zone 24 SM or 258.0 SF

Prevailing Winds

Design Concepts – WHO Level 3 CENTRAL LABORATORY 1

For COLD CLIMATES


For WARM CLIMATES


CULTURE LAB

MDR TESTING LAB


CULTURE LAB

MDR TESTING LAB

BSC

BSC

HAND WASH SINK

MICROSCOPE TABLES

WET BENCH

BSC

HAND WASH SINK

MICROSCOPE TABLES

WET BENCH

PASS- THRU

BOX & TABLE


WAITING ROOM

RECEIVING ROOM


WAITING ROOM

RECEIVING LAB

DESKS or TABLES

STORAGE CABINETS PASS-

THRU BOX & TABLE

PASS- THRU

BOX

HAND WASH SINK

HAND WASH SINK

LAB ENTRY

LAB ENTRY


MDR TESTING LAB


MDR TESTING LAB

WET BENCH

LAB TABLES

MICROSCOPE TABLE

BSC

BSC

PASS- THRU BOX

LAB ENTRY

HAND WASH SINK

LAB SINK

Design Concepts – WHO Level 3

DNA SEQUENCING LAB

DNA EXTRACTION LAB

CENTRAL LABORATORY 2 PCR AMPLIFICATION LAB

STEP 1

STEP 2

STEP 3

Design Concepts – WHO Level 3

DNA SEQUENCING LAB

DNA EXTRACTION LAB

CENTRAL LABORATORY 2 PCR AMPLIFICATION LAB

2 PASS-THRU TRANSFER BOXES

HAND WASH SINK

HAND WASH SINK HAND

WASH SINK

WET BENCH WET

BENCH

EQUIPMENT BENCH

BSC

REF

EQUIPMENT TABLES

PASS- THRU BOX

PASS- THRU BOX

COMPUTER TABLE

LAB ENTRY

LAB ENTRY

Biosafety Laboratories: Containment vs Exclusion Planning & Design Considerations

Janet Baum, AIA

Washington University in St. Louis

biosafety laboratories: containment vs exclusion · biosafety level 1 laboratory (bsl-1) • basic...

Documents