Bio-containment

A subsidiary of GRG Inc.

Differences inBSL-2, BSL-3, BSL-3Ag and

BSL-4Labs and Animal Facilities

Michael C. Connor, PEPrincipal

Earl Walls Associates

*CDC-NIH Biosafety in Microbiology and Biomedical Laboratories, 4th edition

Biosafety Level 2 (BSL-2)*. Used with agents of moderate potential hazard to personnel, animals, and the environment, with minimal economic loss to the animal industries

Wall Construction: Gypsum Board or Masonry

Ceiling Construction: Lay-in acoustic tile with vinyl surface

BSL-2 Laboratory

BSL-2 Laboratory

• Design commissioning – verify designed access to various commissioning points of measurement –Volume Control Boxes don’t conflict with cable trays, etc.

• Verify air flows in accordance with air flow schedules.

• Verify space pressurization.

• Verify access to critical components – Pressure and air flow transmitters.

• Verify labeling of all components requiring maintenance.

BSL-2 Laboratory

• Verify HEPA filter integrity in BSC’s by particle count within cabinet.

• Verify installation of any and all other scientific equipment like point of use DI water systems and centrifuges.

• Perform ASHRAE 110 testing on chemical fume hoods.

• Upset test control system – input set point at edge of range and verify ability to return to normal set point within reasonable time period.

• Verify electrical outlets – voltage, polarity, labeling outlets to panel board circuits.

• Verify shunt trip of lab electrical panels.

• Calibrate all HVAC sensors – flow, pressure, temperature, humidity – don’t depend on factory calibration.

• TRAINING – have mechanics responsible for maintenance participate in commissioning process – most important step and most overlooked.

• Document – video tape training sessions, preserve commissioning baseline.

Biosafety Level 3 (BSL-3)* is applicable to clinical,

diagnostic, teaching, research or production facilities

In which work is done with indigenous or exotic

agents which may cause serious or potentially lethal

disease as a result of exposure by the inhalation route.

*CDC-NIH Biosafety in Microbiology and Biomedical Laboratories, 4th edition

Wall Construction: Clean Room panel (BSL-3), Masonry (BSL-3+) or Poured Concrete (BSL-3++)

Ceiling Construction: Clean Room Gasketed Panel System (BSL-3) or Exposed Structure (BSL-3+) or Coated Structure (BSL-3++)

Example of Panel System

Wall Construction: Masonry

BSL-3 Laboratory

BSL-3 Laboratory

BSL-3 Laboratory

• All of the items covered in BSL-2.

• Autoclave qualification.

• Inspect room penetrations and seals – smoke test.

• Verify VHP effectiveness.

• Calibrate local pressure indicators

• Security system qualification

• NSF 49 - 2004 test of BSC’s cabinets

http://www.afm.ars.usda.gov/ppweb/242-01m.htm#H308

USDA-ARS and APHIS

ZOONOTIC DISEASES are diseases caused by infectious agentsthat can be transmitted between (or are shared by) animals and humans.

Biosafety Level 3 Agriculture (BSL-3Ag). Used with pathogens that present a risk of causing infections of animals and plants and causing a great economic harm. (Foot and Mouth Disease is the premier example.)

Wall Construction: Poured Concrete with Coating

Ceiling Construction: Exposed Structure with Coating

BSL-3ag Laboratory

BSL-3ag Laboratory

BSL-3ag Laboratory• All of the requirements for BSL-2 and BSL-3• Autoclave temperature profile.

– Place typical load in autoclave– Thermocouple mapping of chamber with multi channel recorder.

• Air Pressure Resistant (APR) door verification.– Air Compressor– Air Quality

• Wall Coating Testing– Pull-off testing– Spark Testing

• Inspect utility penetrations– Visually inspect BOTH sides of penetrations.

• Helium leak testing of Ducts and HEPA housings– Outboard test– 1 x 10 –5 cc/sec acceptance criteria. 1 x 10-6 cc/sec atmospheric

background level – requires mass spectrometer NOT thermal conductivity equipment to sense at this level.

BSL-3ag Laboratory

• Space Pressure Decay Testing

– Room must hold 500 Pa (+/-) with a maximum decay to 250 Pa in 20 minutes.

– Epoxy Seal conduits with wiring.

– Impervious wall surfaces.

• HEPA Filter challenges per BSL-3.

• Helium leak testing of ductwork between laboratory and HEPA filters.

– All welded ductwork required – increase wall thickness.

– Outboard testing.

BSL-3ag Laboratory

• Pressure decay testing of double wall plumbing drain pipe

– Test Annular space

– Hold negative pressure – limit to 5 psi MAX

• Waste treatment plant endotoxin challenge – 6 log reduction of Colony Forming Units (CFU), heat resistant bacteria (Bacillus Steareothermophillus)used for challenge.

POWERED AIR PURIFYING RESPIRATOR (PAPR)

WATER SHOWER

CHEMICAL SHOWER

BSL-4

CHEMICAL TANK

BSL-4

• Same as BSL-3ag

• Commissioning Breathing Air System

• Commissioning Chemical Shower System

BSL - 4

BSL - 4

American Biological Safety Association web sitehttp://www.absa.org/resriskgroup.html

Information resources:

CDC-NIH Biosafety in Microbiology and Biomedical Laboratories, 4th editionhttp://des.od.nih.gov/

NIH Design and Policy Guidelineshttp://des.od.nih.gov/eWeb/planning/htm/nihpol.htm

USDA/ ARS/ APHIShttp://www.afm.ars.usda.gov/ppweb/242-01m.htm#H308