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Biocontainment Practices Inside the Animal Lab Guglielmo Vismara

Biocontainment Practices Inside the Animal Lab Guglielmo Vismara. Topics Biological Risk Primary and Secondary Barriers Biosafety Cabinets and Decon Layouts. Biocontainment. Biological risk : risk associated to biological agents Microorganisms , Animals manipulation , Allergens…

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Biocontainment Practices Inside the Animal Lab Guglielmo Vismara

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  1. Biocontainment Practices Inside the Animal Lab Guglielmo Vismara

  2. Topics • BiologicalRisk • Primary and SecondaryBarriers • BiosafetyCabinetsand Decon • Layouts

  3. Biocontainment • Biologicalrisk: riskassociatedtobiologicalagents • Microorganisms, Animalsmanipulation, Allergens… • Invisiblerisk, itcouldbeunderestimated • Itmustbeassessedbyexperienced and responsible people (biosafetyofficer), forevaluationof the optimalworkingconditions • Risk = probabilityofaccident x consequenceofaccident • The riskisalwayspresent. It can beminimizedbutnevereliminated at all • Pathogensassociatedwithliving animalsincrease the levelofbiologicalrisk

  4. (Animal) Biological Safety Levels ABSL-1 Defined organisms Not known to cause disease in healthy adults Basic Facility ABSL-2 Moderate-risk agents present Disease of varying severity ABSL-3 Indigenous or exotic agents, aerosol transmission Serious and potentially lethal infection Containment Facility ABSL-4 Dangerous or exotic high risk agents, Aerosol transmission Serious and potentially lethal infection

  5. The importance to know what pathogen is in use • MicobacteriumTuberculosis: level 3 (CDC-NIH) • AvianInfluenzaeH5N1: level 2/3 (CDC), level 3 (NIH) • Humanadherentcancercells: generallylevel 2 • - Herpes, Papilloma, Citomegalovirus: level 2

  6. Risk Assesment and Management Evaluationof the risk Multiple Factors Managmentof the risk Risk assessment Risk management Validation and surveillance Correctiveactions

  7. Risk Assesment – examples of factors Evaluationofexposedpersonnel: whoisexposedtoanimals, whoenters the vivarium, who do the service, whoclean the rooms. P.I. – Animal care takers – Technicians, ….. Expositionto the biologicalagent: skincontact, eyescontact, inhalation, ingestion, inoculation. Splashes – Aerosols – Puntures – Animalbites and scratches,… Evaluationof the vivariumrooms layout and all the availableequipment (barriers)

  8. Assesment/Management - examples • Example: contaminatedferret urine spilled on the floor • Is the microorganism in the urine? • Can the urine beinfective? • Whatis the volume ofcontaminant? • Choiceof the absorbent material • Whatwaste container tobeused • What personal protectiveequipmentto wear • Whattoolstousetoremove the waste • Howto decontaminate the floorsurface

  9. Biological Agent Host Risk Environment Managing the Biological Risk • To involve all the aspects in the labactivities • Equipmentchoice, and best use • Organizationof the roomlayouts • Organizationoflogistic in the room • Evaluate the people and material flow in the vivarium • Write and “publish” the SOPs • Trainof people • AllthisprocessisDYNAMIC and subjectedtorevisions

  10. Enclosure for the Biological Agents • Primarybarrier: • first protectivebarrierfrombiohazardto the operators • toprotect people and environmentcloseto the source ofcontamination • physicalshield • Biosafety cabinet, animalventilated/sealedcage, tube, centrifuge… • EveryequipmentmustbeassociatedwithSOPstobeaneffectivebarrier

  11. Primary barriers When working with an infected animal, it is the cage

  12. Features of a Good Primary Barrier • Tight Sealed, no air leakage • FiltrationbyHEPAfilters – high efficiency, biologicalfilters • Autoclavable • Compatiblewithdisinfectants and chemicals • Compatiblewith the sizeof a Biosafety Cabinet • Ventilationcontrolledbypressure • Air proof – Water proof

  13. Personal Protective Equipments - If animals are housed in open cages, PPEs are the primary barrier - In case of sealed primary enclosures, PPEs can be downgraded, anyway always represent an important protection PPEs: • Gloves • Coats/Gowns • Respirators • Goggles • Face shields • Shoe covers

  14. Secondary barriers Depend on the transmission risk of agents used. The design, engineering and construction of the facility provides protection for the laboratory workers’ & provides a barrier to protect persons outside the laboratory.

  15. ABSL3vivarium: what’s “HOT”??? HEPA AUTOCLAVE MICROISOLATORS? OPEN CAGES? IVCs’? Cages on Rack ClassicChanging station

  16. BSL3vivarium: what’s “HOT”??? HEPA AUTOCLAVE MICROISOLATORS? OPEN CAGES? IVCs’? • Area fully contaminated • PPE Risk assessment Cages on Rack Laminar Flow

  17. BSL3vivarium: what’s “HOT”??? HEPA AUTOCLAVE Biocontainment at cagelevel Laminar Flow

  18. BSL3vivarium: what’s “HOT”??? HEPA AUTOCLAVE Contamination at enclosures level PPE Butdifferentriskassessment

  19. Some peculiarities of a Biocontainment Lab • Equipment • Changing station = Biosafety Cabinet (ClassII, Biohazard…) • Animalcage = sealedcontainmentcage • PPE = verywelldefined, oftenrespirators • Rooms • Veryrestrictedaccess, lockeddoors • Negative pressure, constantmonitoring • Decontaminate allistaken out • Anteroom, gradientsofpressure, showers • Strong Procedures

  20. Biosafety Cabinets • Veryimportantequipmentforprocedures/animalcagechange/biologicalagentsmanipulation • MandatoryforBSL and ABSL > 1 • Certifiedequipment, requires training and abilitytouse • Laminar Flow, protectsproduct, people and environment under specificcertifications

  21. Biosafety Cabinets • ClassII Cabinet – EN12469, NSF49 • Frontal aperture, the operator can manipulate the product inside the cabinet • Frontal air barriertoprotect the operator • Laminar flow from the top toprotect the product • HEPAfilteredexhausted air toprotect the environment • Partialrecirculationof air (60-70%)

  22. Decontamination Different words around this topic Sanitization: reduction of the microorganisms up to a level considered safe for public health Disinfection: chemical or physical process to eliminate a biological agent Sterilization: eliminates-inactivates all the biological agents In a Biocontainment lab/facility the decon equipment is on the barrier

  23. Decontamination • What is the microorganism (virus, bacteria, spore) • Need effective decontamination • What is the media: surface/liquid waste/room • Right concentration and volume • Is the material resistant or not to the treatment • Chemical or physical treatment • Huge choice of chemicals available on the market • Don’t be disoriented!

  24. Decontamination by gas Cold sterilization: useful for what cannot be treated by heat or liquid Rooms: vaporization and ventilation, routine vivarium shutdown Equipment: bulky, not removable from the room, or sensitive (electrics) Performed in decon-locks: flexible system Peroxide – Formaldehide – Clorine Dioxide MUST KNOW: NOT EFFECTIVE BELOW THE SURFACE!

  25. Some BSL3 Layouts • Primary barriers: • Similar to BSL-2 personal protective equipment • Respiratory equipment if risk of infection through inhalation • Secondary barriers: • Autoclave in the barrier • Corridors separated from direct access to lab • Access through self-closing double doors, locked • Air handling systems to ensure negative air flow (air flows into the lab) • BSC II or BSC III

  26. Some BSL3 Layouts BSL 3

  27. Guidelines • BMBL 5th Edition (CDC/NIH)

  28. Guidelines

  29. Thanks for your attention!

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