Differences in BSL-2, BSL-3, BSL-3Ag and BSL-4 Labs and Animal Facilities Michael C. Connor, PE Principal Earl Walls Associates
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
*CDC-NIH Biosafety in Microbiology and Biomedical Laboratories, 4th edition
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
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
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.) ZOONOTIC DISEASES are diseases caused by infectious agents that can be transmitted between (or are shared by) animals and humans.
USDA-ARS and APHIS http://www.afm.ars.usda.gov/ppweb/242-01m.htm#H308
Wall Construction: Poured Concrete with Coating
Ceiling Construction: Exposed Structure with Coating
BSL-3ag Laboratory
BSL-3ag Laboratory
BSL-3ag Laboratory • •
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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
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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)
BSL-4
WATER SHOWER CHEMICAL SHOWER
CHEMICAL TANK
BSL-4 • Same as BSL-3ag • Commissioning Breathing Air System • Commissioning Chemical Shower System
BSL - 4
BSL - 4
Information resources: American Biological Safety Association web site http://www.absa.org/resriskgroup.html CDC-NIH Biosafety in Microbiology and Biomedical Laboratories, 4th edition http://des.od.nih.gov/ NIH Design and Policy Guidelines http://des.od.nih.gov/eWeb/planning/htm/nihpol.htm USDA/ ARS/ APHIS http://www.afm.ars.usda.gov/ppweb/242-01m.htm#H308
