General Requirements General Requirements for a BSL-3/4 Laboratoryfor a BSL-3/4 Laboratory

Mircea Panait, Ovidiu Mircea Panait, Ovidiu Ciungradi, Adrian Onu, Dorel Ciungradi, Adrian Onu, Dorel

Lucian RaduLucian Radu

Classification of infective Classification of infective microorganisms by risk groupmicroorganisms by risk group

► Risk group 1Risk group 1 ( (no or very low individual and community riskno or very low individual and community risk)) A microorganism that is unlikely to cause human or animal disease.A microorganism that is unlikely to cause human or animal disease.

► Risk group 2Risk group 2 ( (moderate individual risk, low community riskmoderate individual risk, low community risk)) A pathogen that can cause human or animal disease but is unlikely to be 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 a serious hazard to laboratory workers, the community, livestock or the environment.environment.

Laboratory exposures may cause serious infection, but effective Laboratory exposures may cause serious infection, but effective treatment and preventive measures are available and the risk of spread treatment and preventive measures are available and the risk of spread of infection is limited.of infection is limited.

► Risk group 3Risk group 3 ( (high individual risk, low community riskhigh individual risk, low community risk)) A pathogen that usually causes serious human or animal disease but A pathogen that usually causes serious human or animal disease but

does not ordinarily spread from one infected individual to another.does not ordinarily spread from one infected individual to another. Effective treatment and preventive measures are available.Effective treatment and preventive measures are available.

► Risk group 4Risk group 4 ( (high individual and community riskhigh individual and community risk)) A pathogen that usually causes serious human or animal disease and A pathogen that usually causes serious human or animal disease and

that can be readily transmitted from one individual to another, directly that can be readily transmitted from one individual to another, directly or indirectly.or indirectly.

Effective treatment and preventive measures are not usually available.Effective treatment and preventive measures are not usually available.

National or regional classification of National or regional classification of microorganisms by risk groupmicroorganisms by risk group

► Pathogenicity of the organismPathogenicity of the organism► Mode of transmission and host range of the organismMode of transmission and host range of the organism

Existing levels of immunity in the local populationExisting levels of immunity in the local population Density and movement of the host populationDensity and movement of the host population Presence of appropriate vectorsPresence of appropriate vectors Standards of environmental hygieneStandards of environmental hygiene

► Local availability of effective preventive measuresLocal availability of effective preventive measures Prophylaxis by immunization or administration of antiseraProphylaxis by immunization or administration of antisera Sanitary measures (food and water hygiene)Sanitary measures (food and water hygiene) Control of animal reservoirs or arthropod vectorsControl of animal reservoirs or arthropod vectors

► Local availability of effective treatmentLocal availability of effective treatment Passive immunizationPassive immunization Post exposure vaccinationPost exposure vaccination Use of antimicrobial, antiviral and chemotherapeutic agentsUse of antimicrobial, antiviral and chemotherapeutic agents

Relation of risk groups to biosafety Relation of risk groups to biosafety levels, practices and equipmentlevels, practices and equipment

Risk Risk GrouGrou

pp

Biosafety Biosafety LevelLevel

Laboratory Laboratory TypeType

Laboratory Laboratory PracticesPractices

Safety Safety EquipmentEquipment

11Basic – Basic –

Biosafety Level Biosafety Level 11

Basic teaching, Basic teaching, researchresearch GMTGMT None; open bench None; open bench

workwork

22Basic – Basic –

Biosafety Level Biosafety Level 22

Primary health Primary health services; services;

diagnostic, diagnostic, researchresearch

GMT plus GMT plus protective protective clothing, clothing,

biohazard signbiohazard sign

Open bench plus Open bench plus BSC for potential BSC for potential

aerosolsaerosols

33Containment – Containment – Biosafety Level Biosafety Level

33

Special Special diagnostic, diagnostic, researchresearch

As Level 2 plus As Level 2 plus special clothing, special clothing,

controlled controlled access, access,

directional air directional air flowflow

BSC and/or other BSC and/or other primary devices for primary devices for

all activitiesall activities

44

Maximum Maximum containment – containment – Biosafety Level Biosafety Level

44

Dangerous Dangerous pathogen unitspathogen units

As Level 3 plus As Level 3 plus airlock entry, airlock entry, shower exit, shower exit,

special waste special waste disposaldisposal

Class III BSC, or Class III BSC, or positive pressure positive pressure

suits in conjunction suits in conjunction with Class II BSCs, with Class II BSCs,

double-ended double-ended autoclave (through autoclave (through the wall), filtered airthe wall), filtered air

Summary of biosafety level Summary of biosafety level requirements (1)requirements (1)

Biosafety levelBiosafety level

11 22 33 44

Isolation of laboratoryIsolation of laboratory NoNo NoNo DesirablDesirablee

YesYes

Room sealable for decontaminationRoom sealable for decontamination NoNo NoNo YesYes YesYes

Ventilation:Ventilation:

► inward air flowinward air flow NoNo DesirablDesirablee

YesYes YesYes

► mechanical via building systemmechanical via building system NoNo DesirablDesirablee

YesYes NoNo

► mechanical, independentmechanical, independent NoNo DesirablDesirablee

YesYes YesYes

► HEPA filtered air exhaustHEPA filtered air exhaust NoNo NoNo DesirablDesirablee

YesYes

Double-door entryDouble-door entry NoNo NoNo YesYes YesYes

AirlockAirlock NoNo NoNo NoNo YesYes

Airlock with showerAirlock with shower NoNo NoNo NoNo YesYes

AnteroomAnteroom NoNo NoNo YesYes NoNo

Anteroom with showerAnteroom with shower NoNo NoNo DesirablDesirablee

NoNo

Effluent treatmentEffluent treatment NoNo NoNo DesirablDesirablee

YesYes

Summary of biosafety level Summary of biosafety level requirements (2)requirements (2)

Biosafety levelBiosafety level

11 22 33 44

Autoclave:Autoclave:

► on siteon site YesYes YesYes YesYes YesYes

► in laboratory roomin laboratory room NoNo NoNo DesirableDesirable YesYes

► double-endeddouble-ended NoNo NoNo DesirableDesirable YesYes

Biological safety cabinets:Biological safety cabinets:

► Class IClass I NoNo OptionalOptional YesYes NoNo

► Class IIClass II NoNo DesirableDesirable YesYes

Yes, in Yes, in conjunctioconjunction with suit n with suit laboratorielaboratorie

ss

► Class IIIClass III NoNo NoNo DesirableDesirable

Yes, in Yes, in conjunctioconjunctio

n with n with cabinet cabinet

laboratorielaboratoriess

BSL-4 projectBSL-4 project

BSL-4 project or facility can operate correctly BSL-4 project or facility can operate correctly and in safe condition if we have:and in safe condition if we have:

► A good design, engineering, construction and A good design, engineering, construction and validation of the installation.validation of the installation.

► A trained personnel and good procedure system.A trained personnel and good procedure system.► A good maintenance team which ensure the well A good maintenance team which ensure the well

state of the installation.state of the installation.

General considerations General considerations regarding BSL-4 laboratory – regarding BSL-4 laboratory –

from the outsidefrom the outsideBSL-4 design features include the following:BSL-4 design features include the following:► All seams, joints and doors are sealed to make the building airtight. 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 You can not open a window. Air does not flow in or out under the doors.doors.

► Air is pumped in and out of the building through a filtration system Air is pumped in and out of the building through a filtration system that catches the tiniest microscopic particles, including bacteria and that catches the tiniest microscopic particles, including bacteria and viruses.viruses.

► All air ducts are welded stainless steel and tested to be airtight.All air ducts are welded stainless steel and tested to be airtight.► Inside many BSL-4 facilities, laboratories are surrounded by buffer Inside many BSL-4 facilities, laboratories are surrounded by buffer

corridors that help protect the labs in the event of bombing attack corridors that help protect the labs in the event of bombing attack from outside.from outside.

► Features such airlocks, fumigation chambers, disinfectant “dunk Features such airlocks, fumigation chambers, disinfectant “dunk tanks” and waste water treatment systems ensure that absolutely tanks” and waste water treatment systems ensure that absolutely everything that leaves a BSL-4 laboratory is decontaminated.everything that leaves a BSL-4 laboratory is decontaminated.

1.1. A security checkpoint at the entry to the siteA security checkpoint at the entry to the site2.2. A secured perimeter around the buildingA secured perimeter around the building3.3. A buffer between the building and parking areas, drop-off zonesA buffer between the building and parking areas, drop-off zones4.4. On-site 24-hour surveillanceOn-site 24-hour surveillance5.5. An additional vehicle checkpoint for lab employees, visitorsAn additional vehicle checkpoint for lab employees, visitors6.6. A secured entryway, including security guards, surveillance cameras, an A secured entryway, including security guards, surveillance cameras, an

intercom system and card access readersintercom system and card access readers7.7. A delivery vehicle inspection areaA delivery vehicle inspection area8.8. The building also contains numerous safety and security measures The building also contains numerous safety and security measures

throughoutthroughout

General considerations General considerations regarding BSL-4 laboratory – regarding BSL-4 laboratory –

interior securityinterior security► Once pass outside security, BSL-4 lab workers must Once pass outside security, BSL-4 lab workers must

pass an interior security stationpass an interior security station► Security measures will include combinations of the Security measures will include combinations of the

following:following: Extensive background checks prior to employment for Extensive background checks prior to employment for

everyone who works in the buildingeveryone who works in the building Guards who check everyone’s identification and credentialsGuards who check everyone’s identification and credentials Special identification cards that are scanned to provide an Special identification cards that are scanned to provide an

electronic record of who goes in and out, and whenelectronic record of who goes in and out, and when Identifications devices such as retinal or fingerprint scannersIdentifications devices such as retinal or fingerprint scanners

Scouring the air – Scouring the air – HEPA filter and filtration inspection HEPA filter and filtration inspection

verificationverification

Changing roomChanging room

Inside the labInside the lab► Entries and exits have double-door airlocksEntries and exits have double-door airlocks► Work surfaces are regularly decontaminatedWork surfaces are regularly decontaminated► Researchers are vaccinated against the pathogen they are studyingResearchers are vaccinated against the pathogen they are studying► All solid and liquid waste is decontaminated by heat sterilization, All solid and liquid waste is decontaminated by heat sterilization,

gaseous sterilization or liquid disinfectantgaseous sterilization or liquid disinfectant

Biocontainment Suits and Biosafety Biocontainment Suits and Biosafety CabinetsCabinets

► The airtight, pressurized suits have dedicated life support systems that The airtight, pressurized suits have dedicated life support systems that include air compressors, alarms and emergency backup air tanks and include air compressors, alarms and emergency backup air tanks and a HEPA air filtration systema HEPA air filtration system

► Work stations called biosafety cabinets serve as additional barriersWork stations called biosafety cabinets serve as additional barriers► Different type of biosafety cabinet can be used (Class II or III)Different type of biosafety cabinet can be used (Class II or III)

Decontaminating ShowerDecontaminating Shower

Community and EnvironmentCommunity and Environment