General Requirements for a BSL-3/4 Laboratory Mircea Panait, Ovidiu Ciungradi, Adrian Onu, Dorel Lucian Radu

Classification of infective microorganisms by risk group Risk group 1 (no or very low individual and community risk) A microorganism that is unlikely to cause human or animal disease. Risk group 2 (moderate individual risk, low community risk) A pathogen that can cause human or animal disease but is unlikely to be a serious hazard to laboratory workers, the community, livestock or the environment. Laboratory exposures may cause serious infection, but effective treatment and preventive measures are available and the risk of spread of infection is limited. Risk group 3 (high individual risk, low community risk) A pathogen that usually causes serious human or animal disease but does not ordinarily spread from one infected individual to another. Effective treatment and preventive measures are available. Risk group 4 (high individual and community risk) A pathogen that usually causes serious human or animal disease and that can be readily transmitted from one individual to another, directly or indirectly. Effective treatment and preventive measures are not usually available.

National or regional classification of microorganisms by risk group Pathogenicity of the organism Mode of transmission and host range of the organism Existing levels of immunity in the local population Density and movement of the host population Presence of appropriate vectors Standards of environmental hygiene Local availability of effective preventive measures Prophylaxis by immunization or administration of antisera Sanitary measures (food and water hygiene) Control of animal reservoirs or arthropod vectors Local availability of effective treatment Passive immunization Post exposure vaccination Use of antimicrobial, antiviral and chemotherapeutic agents

Relation of risk groups to biosafety levels, practices and equipment Laboratory Type Laboratory Practices Safety Equipment 1 Basic – Biosafety Level 1 Basic teaching, research GMT None; open bench work 2 Basic – Biosafety Level 2 Primary health services; diagnostic, research GMT plus protective clothing, biohazard sign Open bench plus BSC for potential aerosols 3 Containment – Biosafety Level 3 Special diagnostic, research As Level 2 plus special clothing, controlled access, directional air flow BSC and/or other primary devices for all activities 4 Maximum containment – Biosafety Level 4 Dangerous pathogen units As Level 3 plus airlock entry, shower exit, special waste disposal Class III BSC, or positive pressure suits in conjunction with Class II BSCs, double-ended autoclave (through the wall), filtered air

Summary of biosafety level requirements (1) 2 3 4 Isolation of laboratory No Desirable Yes Room sealable for decontamination Ventilation: inward air flow mechanical via building system mechanical, independent HEPA filtered air exhaust Double-door entry Airlock Airlock with shower Anteroom Anteroom with shower Effluent treatment

Summary of biosafety level requirements (2) 1 2 3 4 Autoclave: on site Yes in laboratory room No Desirable double-ended Biological safety cabinets: Class I Optional Class II Yes, in conjunction with suit laboratories Class III Yes, in conjunction with cabinet laboratories

BSL-4 project BSL-4 project or facility can operate correctly and in safe condition if we have: A good design, engineering, construction and validation of the installation. A trained personnel and good procedure system. A good maintenance team which ensure the well state of the installation.

General considerations regarding BSL-4 laboratory – from the outside BSL-4 design features include the following: All seams, joints and doors are sealed to make the building airtight. You can not open a window. Air does not flow in or out under the doors. Air is pumped in and out of the building through a filtration system that catches the tiniest microscopic particles, including bacteria and viruses. All air ducts are welded stainless steel and tested to be airtight. Inside many BSL-4 facilities, laboratories are surrounded by buffer corridors that help protect the labs in the event of bombing attack from outside. Features such airlocks, fumigation chambers, disinfectant “dunk tanks” and waste water treatment systems ensure that absolutely everything that leaves a BSL-4 laboratory is decontaminated.

A security checkpoint at the entry to the site A secured perimeter around the building A buffer between the building and parking areas, drop-off zones On-site 24-hour surveillance An additional vehicle checkpoint for lab employees, visitors A secured entryway, including security guards, surveillance cameras, an intercom system and card access readers A delivery vehicle inspection area The building also contains numerous safety and security measures throughout

General considerations regarding BSL-4 laboratory – interior security Once pass outside security, BSL-4 lab workers must pass an interior security station Security measures will include combinations of the following: Extensive background checks prior to employment for everyone who works in the building Guards who check everyone’s identification and credentials Special identification cards that are scanned to provide an electronic record of who goes in and out, and when Identifications devices such as retinal or fingerprint scanners

Scouring the air – HEPA filter and filtration inspection verification

Changing room

Inside the lab Entries and exits have double-door airlocks Work surfaces are regularly decontaminated Researchers are vaccinated against the pathogen they are studying All solid and liquid waste is decontaminated by heat sterilization, gaseous sterilization or liquid disinfectant

Biocontainment Suits and Biosafety Cabinets The airtight, pressurized suits have dedicated life support systems that include air compressors, alarms and emergency backup air tanks and a HEPA air filtration system Work stations called biosafety cabinets serve as additional barriers Different type of biosafety cabinet can be used (Class II or III)

Decontaminating Shower

Community and Environment