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Biocontainment Practices Inside the Animal Lab Guglielmo Vismara 1.

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Presentation on theme: "Biocontainment Practices Inside the Animal Lab Guglielmo Vismara 1."— Presentation transcript:

1 Biocontainment Practices Inside the Animal Lab Guglielmo Vismara 1

2 Topics - Biological Risk - Primary and Secondary Barriers - Biosafety Cabinets and Decon - Layouts 2

3 Biocontainment - Biological risk: risk associated to biological agents Microorganisms, 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 3

4 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-4 Dangerous or exotic high risk agents, Aerosol transmission Serious and potentially lethal infection Basic Facility Containment Facility 4 (Animal) Biological Safety Levels

5 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 5

6 Risk Assesment and Management Risk assessment Risk management Evaluation of the risk Managment of the risk Validation and surveillance Corrective actions Multiple Factors

7 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)

8 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

9 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 Biological Agent Host Environment Risk

10 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 10

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

12 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 12

13 - 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 13 Personal Protective Equipments

14 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. 14 Secondary barriers

15 ABSL3 vivarium: whats HOT??? HEPA AUTOCLAVE Cages on Rack Classic Changing station MICROISOLATORS? OPEN CAGES? IVCs?

16 HEPA AUTOCLAVE Cages on Rack Laminar Flow MICROISOLATORS? OPEN CAGES? IVCs? - Area fully contaminated - PPE Risk assessment BSL3 vivarium: whats HOT???

17 HEPA AUTOCLAVE Laminar Flow Biocontainment at cage level BSL3 vivarium: whats HOT???

18 HEPA AUTOCLAVE Contamination at enclosures level PPE But different risk assessment BSL3 vivarium: whats HOT???

19 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

20 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

21 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%)

22 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

23 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 Dont be disoriented! Decontamination

24 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

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 27

28 Guidelines BMBL 5 th Edition (CDC/NIH)

29 Guidelines

30 Thanks for your attention!


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