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Biocontainment Practices Inside the Animal Lab
Guglielmo Vismara
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Topics
- Biological Risk
- Primary and Secondary Barriers
- Biosafety Cabinets and Decon
- Layouts
Biocontainment
- Biological risk: risk associated to biological agentsMicroorganisms, Animals manipulation, Allergens…
- Invisible risk, it could be underestimated
- It must be assessed by experienced and responsible people (biosafety officer), for evaluation of the optimal working conditions
- Risk = probability of accident x consequence of accident
- The risk is always present. It can be minimized but never eliminated at all
- Pathogens associated with living animals increase the level of biological risk
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ABSL-1 Defined organisms Not known to cause disease in healthy adults
ABSL-2 Moderate-risk agents present Disease of varying severity
ABSL-3 Indigenous or exotic agents, aerosol transmission Serious and potentially lethal infection
ABSL-4Dangerous or exotic high risk agents, Aerosol transmission Serious and potentially lethal infection
Basic Facility
Containment Facility
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(Animal) Biological Safety Levels
The importance to know what pathogen is in use
- Micobacterium Tuberculosis: level 3 (CDC-NIH) - Avian Influenzae H5N1: level 2/3 (CDC), level 3 (NIH)
- Human adherent cancer cells: generally level 2
- Herpes, Papilloma, Citomegalovirus: level 2
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Risk Assesment and Management
Risk assessment
Risk management
Evaluation of the risk
Managment of the risk
Validation and surveillance
Corrective actions
Multiple Factors
Risk Assesment – examples of factors
1) Evaluation of exposed personnel: who is exposed to animals, who enters the vivarium, who do the service, who clean the rooms. P.I. – Animal care takers – Technicians, …..
2) Exposition to the biological agent: skin contact, eyes contact, inhalation,
ingestion, inoculation. Splashes – Aerosols – Puntures – Animal bites and scratches,…
3) Evaluation of the vivarium rooms layout and all the available equipment(barriers)
Assesment/Management - examples
Example: contaminated ferret urine spilled on the floor
- Is the microorganism in the urine?
- Can the urine be infective?
- What is the volume of contaminant?
- Choice of the absorbent material
- What waste container to be used
- What personal protective equipment to wear
- What tools to use to remove the waste
- How to decontaminate the floor surface
Managing the Biological Risk
To involve all the aspects in the lab activities
- Equipment choice, and best use
- Organization of the room layouts
- Organization of logistic in the room
- Evaluate the people and material flow in the vivarium
- Write and “publish” the SOPs
- Train of people
All this process is DYNAMIC and subjected to revisions
BiologicalAgent Host
Environment
Risk
Enclosure for the Biological Agents
Primary barrier:
- first protective barrier from biohazard to the operators- to protect people and environment close to the source of contamination- physical shield Biosafety cabinet, animal ventilated/sealed cage, tube, centrifuge…
Every equipment must be associated with SOPs to be an effective barrier
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Primary barriers
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When working with an infected animal, it is the cage
Features of a Good Primary Barrier
- Tight Sealed, no air leakage
- Filtration by HEPA filters – high efficiency, biological filters
- Autoclavable
- Compatible with disinfectants and chemicals
- Compatible with the size of a Biosafety Cabinet
- Ventilation controlled by pressure
- Air proof – Water proof
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- 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
Personal Protective Equipments
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The design, engineering and construction of the facility provides protection for the laboratory workers’ & provides a barrier to protect persons outside the laboratory.
Depend on the transmission risk of agents used.
Secondary barriers
ABSL3 vivarium: what’s “HOT”???
HEPA AUTOCLAVE
Cage
s on
Rac
k
Classic Changing station
MICROISOLATORS?OPEN CAGES?IVCs’?
HEPA AUTOCLAVE
Cage
s on
Rac
k
Laminar Flow
MICROISOLATORS?OPEN CAGES?IVCs’?
- Area fully contaminated- PPE
Risk assessment
BSL3 vivarium: what’s “HOT”???
HEPA AUTOCLAVE
Laminar Flow
Biocontainment at cage level
BSL3 vivarium: what’s “HOT”???
HEPA AUTOCLAVE
Contamination at enclosures levelPPEBut different risk assessment
BSL3 vivarium: what’s “HOT”???
Some peculiarities of a Biocontainment Lab
Equipment
- Changing station = Biosafety Cabinet (Class II, Biohazard…)
- Animal cage = sealed containment cage
- PPE = very well defined, often respirators
Rooms
- Very restricted access, locked doors
- Negative pressure, constant monitoring
- Decontaminate all is taken out
- Anteroom, gradients of pressure, showers
Strong Procedures
Biosafety Cabinets
- Very important equipment for procedures/animal cage change/biological agents
manipulation
- Mandatory for BSL and ABSL > 1
- Certified equipment, requires training and ability to use
- Laminar Flow, protects product, people and environment under specific certifications
Biosafety Cabinets
Class II Cabinet – EN12469, NSF49
- Frontal aperture, the operator can manipulate
the product inside the cabinet
- Frontal air barrier to protect the operator
- Laminar flow from the top to protect the product
- HEPA filtered exhausted air to protect the
environment
- Partial recirculation of air (60-70%)
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
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 treatmentChemical or physical treatment
Huge choice of chemicals available on the market Don’t be disoriented!
Decontamination
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!
Decontamination by gas
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
Some BSL3 LayoutsBSL 3
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Guidelines
BMBL 5th Edition (CDC/NIH)
Guidelines
Thanks for your attention!