Human & Animal Health Laboratories: from Concept to Commissioning
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Human & Animal Health Laboratories: from Concept to Commissioning Yanko Ivanov & Ragip Bayraktar, EU Technical Assistance to Avian Influenza Preparedness & Response Project, EU. Contents. Biosafety & Biosecurity considerations and p rinciples

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Human & Animal Health Laboratories: from Concept to CommissioningYanko Ivanov & Ragip Bayraktar,EU Technical Assistance to Avian Influenza Preparedness & Response Project, EU


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Contents Commissioning

  • Biosafety &Biosecurity considerations and principles

  • Assessment for laboratory need and decision –making process and rationale for investments in laboratories or not.

  • Determination of functions and scope of work in a laboratory

  • Commissioning and operations.

  • Design and construction Issues.


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Laboratory biosafety Commissioning & biosecurity - definitions

  • “Laboratory biosafety” is the term used to describe the containment principles,technologies and practices that are implemented to prevent unintentional exposureto pathogens and toxins, or their accidental release.

  • “Laboratory biosecurity” refers toinstitutional and personal security measures designed to prevent the loss, theft, misuse,diversion or intentional release of pathogens and toxins.


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Laboratory biosecurity as a complement Commissioningto laboratory biosafety

  • Laboratory biosafety and biosecurity mitigate different risks, but they share a commongoal: keeping valuable biological materialssafely and securely inside the areas where they are used and stored.

  • Good laboratory biosafety practices reinforce and strengthen laboratory biosecuritysystems.


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Conflicts Commissioning (Biosafety vs Biosecurity)

Accountability

Compliance SOP’s

Emergency routines

GLP

Safety cabinets

Personal safety equipment

Inventory control

Access control

Transfer security

Physical security

Information security

Biosafety

containment

Biosecurity

Incident reporting

Incident response planning

Revision

Training

Supervised by appointed Biosafety officer

Regulated by national work environment safety law

Supervised by appointed Biosecurity or Biosafety officer

Should consult with law enforcement officials and security experts


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Why Commissioningbiosafety practices?

  • Protection:

    - workers

    - “products”

    - co-workers

    - lab support personnel

    - environment


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Biosafety Commissioninglevels

  • BSL1 - agents not known to cause disease (no or low individual and community risk).

  • BSL2 - agents that cause human or animal diseaseswith moderate individual or low community risk (e.g. blood borne diseases).

  • BSL3 - indigenous/exotic agents associatedwith human disease and with potential foraerosol transmission - high individual risk (respiratory) low community risk)

  • BSL4 - dangerous/exotic agents of lifethreatening nature – serious diseases readily transmitted.


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Biosafety Level 3 CommissioningRisk Assessment

  • What is the natural host of the biological agent?

  • Does the agent cross species barriers?

  • Is it a wild-type agent or attenuated?

  • Is the agent infectious for a normal healthy adult?

  • What effect will the agent have on an adult if immunocompromised? if pregnant?


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Biosafety Level 3 CommissioningRisk Assessment

  • What is the mode of transmission for the agent? - contact,

    - mucous membrane exposure,

    - ingestion,

    - inoculation,

    - inhalation

  • What volume of the agent is being manipulated?

  • What is the concentration of the agent?

  • What is the infectious dose of the agent?


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Biosafety Level 3 CommissioningRisk Assessment

  • Prophylaxis

    • What, if any immunizations are required?

    • What pharmaceuticals are available?

    • What is the effectiveness of prophylaxis?

  • Post-exposure

    • What are the anti-microbial agents available for treatment?

    • What is the effectiveness of treatment?



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Summary of biosafety level requirements equipment

BIOSAFETY LEVEL1234

  • Isolation of laboratory No NoYes Yes

  • Room sealable for decontamination No No Yes Yes

  • Ventilation:

    — inward airflow No DesirableYes Yes

    — controlled ventilating system No Desirable Yes Yes

    — HEPA-filtered air exhaust No No Yes/NobYes

  • Double-door entry No No Yes Yes

  • Airlock No No NoYes

  • Airlock with shower No No No Yes

  • Anteroom No No Yes—

  • Anteroom with shower No No Yes/NocNo

  • Effluent treatment No No Yes/NocYes

  • Autoclave:

    — on site No Desirable Yes Yes

    — in laboratory room No No Desirable Yes

    — double-ended No No Desirable Yes

  • Biological safety cabinets No Desirable Yes Yes

  • Personnel safety monitoring capabilityd No No Desirable Yes


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BIOSAFETY AND BIOSECURITY LABORATORY DESIGN CRITERIA equipment

  • Laboratory location

  • Wipe-clean surfaces

  • Heating, ventilation and air-conditioning (HVAC) system

  • Directional airflow and cascade negative pressure

  • Laboratory furniture and equipment

  • Laboratory rooms, size and orientation

  • Sample reception

  • Double door autoclave and decontamination chamber for solid waste materials

  • Water supply and sewerage system

  • Electrical system


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Essential Building Principles equipment

  • Primary containment barrier is the first barrier between agent and man (such as gloves, gowns, masks, biosafety cabinets, respiratory protection etc.)

  • Secondary containmentbarrier is the barrier between agents and environment (airtight rooms, air handling andfiltration, air locks,showers, laundry, sewage treatment, waste disposal, sterilisers, redundantservices as well as equipment and material niches.

  • Tertiary containment barrier represents an additional organisational barrier with the physical operation with items such as walls, fences, security,quarantine and animal exclusion zones.


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Work flow considerations equipment

During the programming phase it is essential to define how various elements are processed, including animals (clean and dirty), people, wastes (carcasses, solid, other), samples from animals, laundry, feed and bedding (if used).


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The containment barrier equipments

The containment barriers should be physical barriers constructed with a series of integrated building components to form an airtight interior environment separate from the surrounding research environment and neighbouring community. The barrier is also to be defined by operational practices – examples of these “secondary barriers” include work areas that are separate from public areas, decontamination, shower and hand-washing procedures and equipment, special ventilation systems, directional airflow through the use of air pressure differentials, double door autoclaves, liquid waste treatment, donning of personal protective equipment (and removal upon exit) and restricted personnel access.


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Specimen Reception, Dispatch Area and "Grey Areas" equipment

  • A good specimen reception would be an isolated containment area, yet a "grey area", for the preliminary handling of diagnostic specimens by experienced pathology personnel. This would ensure the correct movement of samples to laboratories or autopsy.

  • A specimen reception located next to the autopsy area would help to integrate the system of controlled movement of materials into the secure laboratories or animal facilities. A "grey area" would also be an appropriate area to hold reagent awaiting innocuity tests or whilst inactivation is proven.

  • Samples destined for other reference laboratories may be safely removed from the "grey area" without a perception of possible adventitious contamination that might occur if the samples were manipulated in the high containment laboratories before they are removed.


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Security and equipmentrelatedsystems

  • Typically there are various operational zones within containment facilities. Access control to one zone does not necessarily give access control to all rooms or areas within that zone. There are various programs that require individual access control for the appropriate personnel.

  • Fire alarm


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Security Zone 1: Property Protection Area (uncontrolled) equipment

• The entire building should monitor entrance zones with CCTV cameras

• Employee / visitor parking should have CCTV monitoring

• Rear Loading docks should have CCTV monitoring

• The electrical transformer vaults should be secured


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Security Zone 2: Limited Areas / Non-Containment equipment

These areas require a primary level access control credential (proximity card) to enter. Cards should be coded to permit entry into specific Limited Areas based on the need to access.

  • Laboratory corridors (non-containment)

  • Loading dock storage receiving areas and animal delivery airlocks


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Security Zone 3: Exclusion Area / Non-Containment equipment

This areas directly support containment operations and require a third level card access control

  • Housing animal receiving airlock

  • Clean autoclave rooms

  • Basement area where liquid treatment system is located

  • Mechanical penthouse serving all HEPA filters and Air Handling Units, exhaust fans

  • Building operation control areas accommodating building automation systems


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Security Zone 4: Exclusion Areas – Containment equipment

These areas are designated as Secondary Containment Spaces in which the design and access is controlled primarily to allow researchers and operators into the facility. Entry into these areas will require two-level access control, proximity card and PIN and/or biometric. Exit from these areas will require the proximity card. All entrances will have a CCTV camera for monitoring.

• CL3 laboratory shower entrance zone

• CL3 decontamination airlocks


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Security Zone 5: Exclusion Areas – Containment equipment

These areas are designated as Lab Containment Spaces or specialised areas in the design. Entry and exit will require keypad entry of a PIN and/or biometric, which authorises access only to specific modules or spaces. All areas will have CCTV and motion detection.

• CL3 laboratories

• CL3 animal holding rooms


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Security Zone 6: Exclusion Areas – Containment Pathogen Storage Freezers

All freezers should have access control. All rooms are equipped with additional security features including motion detection, door access control, CCTV camera monitoring and special access and use procedures.


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Aerosol Control Storage Freezers

  • Sources of Biohazardous Aerosols

  • Impact of Ventilation on Aerosol Load

  • Air filtration

  • Airlocks

  • Anteroomsas a Control Mechanism

  • Cascade negative pressure

  • SOP and PPE as a Control Mechanism


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Sources of Biohazardous Aerosols Storage Freezers

Biohazardous aerosols of concern in the laboratory setting are generated by a number of manipulations involving infectious material.such assonification, mixing, pouring and pipetting centrifugation, during an accident etc. In animal facilities, aerosols of infectious pathogens may be generated by infected animals breathing, sneezing or coughing respiratory pathogens.


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Ventilation Storage Freezers

Issues related to ventilationin containment facilities include: directional airflow, airflow velocities, pressure differentialbetween adjacent spaces and air exchange rates.

  • Directional airflow is used to create zones of hazard by moving air from areas clear ofhazardous aerosol contamination to areas of higher potential for hazardous aerosolcontamination.This provides for twofunctions:

    - 1) control of the hazardous aerosol minimises the possibility of inadvertentexposure outside of the laboratory space and;

    - 2) knowledge of where the aerosol hazard existsand the extent of the hazard allows personnel to follow appropriate protocols if they arerequired to enter areas where aerosols may exist.


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Air f Storage Freezers iltration

Where the risk assessmentindicates that a significant aerosol release of pathogens outside of primary containment isprobable and would create a hazard to people or the environment outside of the facility, theexhaust system should be HEPA filtered to prevent the release of the pathogens outside of the laboratory.


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Airlocks Storage Freezers

Airlocks have one primary purpose; to eliminate or minimise the transfer of air from the containment zone to a non-containment zone or from one zone or level of containment to another to avoid cross- contamination.

Airlocks, whether it is a PPE room, change room, shower, anteroom, or decontamination chamber (adevice to transfer large pieces of equipment), requires special attention for room tightness, door control and ventilation design. Airlock entry ports for specimens, materials and animals must be available as well.


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Airlocks- cont. Storage Freezers

The airlocks should include the following:

  • Interlocking doors preventing two doors opened at once

  • Directional airflow measurement capability (pressure sensors and alarms)

  • Door swings to accommodate direction of airflow and passage of equipment

  • Direct ventilation of supply and/or exhaust inside airlock depending on many criteria

  • Vision panels in doors unless it’s designated as a change room

  • Tight doors depending on which side and method of controls integration


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Anterooms and two doors in series Storage Freezers

Considerable control of airborne micro-organisms can be achieved withthe addition of ananteroom to the laboratory or animal holding room.

This is the basis for therequirement in BSL-3 or equivalent facilities to have entry by two doors in series.

A laboratory with Class III biosafety cabinets is only accessible through a minimum of two doors.


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Cascade negative pressure Storage Freezers

The pressure decreases at each containment barrier and is lowest at the location of highest potential or effective contamination. For example: security corridor -30 Pa, shower -60 Pa, laboratory -90 Pa, animal room -120 Pa.


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Technical details about pressure differentiation and backflow prevention

  • Pressure differentials in animal facilities are held at approximately 50 Pa lower pressure than the point ofpersonnel entry so that there is airflow into the room upon door opening.

  • Backflow prevention for containment labs is necessary to prevent back siphoning of contaminated liquids and air. Types of backflow solutions are dependent on the medium that is considered: water, air, gas, and steam.


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E backflow preventionlectrical system

The electrical systems of containment laboratories ensure that all of the systems cohesively work together to manage the three essential criteria for biocontainment:

• Protection of the staff

• Protection of scientific programs

• Protection of the environment and adjacent communities

Electrical systems can be segregated into normal power systems, emergency power systems, uninterruptible power systems (UPS), communication systems, data and information systems, lightning control systems, security systems, lighting systems, equipment monitoring systems, automation control systems, life safety systems, harmonic control systems and telemetry systems.


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E backflow preventionmergency power strategy

Emergency power planning for containment facilities does not mean that all loads need to have this provision. It means that critical loads may include life safety, virus collection, sensitive equipment and ventilation systems may be all required. One particular emergency power strategy could be:

• 100% of fire systems *

• 100% of building automation *

• 100% of security *

• 100 % of HVAC (chilling / heating pumps, fans valves)

• 50% of lab receptacles

• 50% of animal room receptacles

• 25% of in-door lighting systems

• 10% of non-lab space

• 10% of outdoor lighting

• 100% of air compressors for containment control

• 0% of compressors for non-containment control

• 100% of all Biological safety cabinets, freezers, incubators

• 100% of all liquid / solid effluent treatment systems


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Emergency Power backflow prevention

Emergency power is needed when there are interruptions or problems with the normal power provided by the utility. The emergency power will allow the facility to continue to operate, usually in a reduced mode feeding only those items considered essential to operate the laboratory and maintain life safety systems. The run time is dictated by the amount of fuel on hand and availability from the suppliers. Fuel storage capacity should ideally be considered for at least 48 hours of operation for a containment facility.


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Identification backflow prevention

Proper identification is extremely important on all systems and equipment. The most expeditious method of handling this would be to consult with the end user to enter their naming convention on the design and construction drawings. This is important when systems are being integrated within existing facilities or where a computerised maintenance management system will be utilised.


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T backflow preventionhe identification should provide information on the following:

• Voltage and Phases

• Type of power, normal emergency or UPS

• Lighting or Power circuits

• Approximate location (e.g. a floor or a wing or building number)

• Short circuit fault current potential at each panel

• Substations (should have a mimic bus on the front of the gear)

• Receptacles (should identify panel and circuit number)

• Switches (line voltage switches should also identify the panel andcircuit number)

• Disconnects / Motor Starters not in an MCC (the source should be indicated, as well as the voltage and the identifier of the load being served)

• Motor Starters in Motor Control Centres (the name of the load that is served)


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Labels backflow prevention

The labels should be colour coded to provide indication of the system. Forexample:

• Normal Power – Black background / white letters.

• Normal Lighting – White background / black letters.

• Emergency Power – Red Background / white letters.

• Emergency Lighting – White Background / red letters.

• UPS Panel – Yellow background / black letters.


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Indicator Lights backflow prevention

Indicator lights should be provided with LEDs (Light Emitting Diodes) as opposed to incandescent lamps wherever possible. The LEDs have a much longer life expectancy than the incandescent lamps providing more reliable indication while consuming significantly less energy. Indicator lights provide a quick assessment of equipment status which is helpful in all situations, especially emergencies.


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Effluent backflow preventiontreatment

  • Heat treatment – 95 C

  • Chemical treatment


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Redundancy backflow prevention

Redundancy is defined as having more than one system supporting an individual mechanical function. It would be wrong to assume that each and every mechanical system or device needs to have redundancy. The primary areas for redundancy need to focus on the three principles of bio-containment- environment protection, personnel protection and product (or scientific outcome) protection. Therefore, during a design process the issue of redundancy needs to be well thought out.


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Laboratory animal facilities backflow prevention

Facilities for laboratory animals used for studies of infectious disease should be physically separated from other activities such as animal production, quarantine and clinical laboratories. As microbiological containment of infected animals is more difficult than for laboratory cultures, animal facilities should be located remotely from experimental laboratories as well. For security reasons, the animal house should be an independent, detached unit. If it adjoins a laboratory, the design should provide for its isolation from the public parts of the laboratory should such need arise, and for its decontamination and disinfestation.


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Planning Experimental Work backflow prevention

  • In an animalbio-containment facility, a basic assumption is that animals will not bring any disease into thefacility that will compromise either the planned experiment or other animals. Therefore animals must be introduced via a path that is free fromdisease agents and that no disease agent will escape from within while fresh animals areintroduced.

  • Access to animal rooms is limited to personnel that have been advised of potential hazards,are trained, meet specific requirements.


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Waste Disposal backflow prevention

  • The safe handling of infectious wastes must be considered as part of the experimental plan.

  • Urine and faecal wastes for animals infected with Level 3 and 4 agents must bedecontaminated either by heat or chemical treatment.

  • Discarded surgery or necropsy tissues from infected animals are usually sterilised byautoclaving and carcasses by rendering at high temperature, steam sterilisation, incineration or chemical decontaminationsuch as alkaline hydrolysis.

  • All infectious wastes that cannot be decontaminated or autoclaved will immediately be placedin red infectious waste bags.


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Laboratory backflow preventioncommissioning

  • Laboratory commissioning may be defined as the systematic review anddocumentation process signifying that specified laboratory structural components,systems and/or system components have been installed, inspected, functionally testedand verified to meet national or international standards, as appropriate.

  • Laboratories designated as Biosafety Levels 1–4 will have different and increasingly complex commissioning requirements

  • The commissioning process and acceptance criteria should be established early, preferably during the programming phase of the construction or renovation project.


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Why backflow preventionlaboratorycommissioning?

The commissioning process provides theinstitution and the surrounding community with a greater degree of confidence thatthe structural, electrical, mechanical and plumbing systems, containment anddecontamination systems, and security and alarm systems will operate as designed, toassure containment of any potentially dangerous microorganisms being worked within a particular laboratory or animal facility.


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L backflow preventionist of laboratory systemsin the commissioning plan

1. Building automation systems including links to remote monitoring and controlsites

2. Electronic surveillance and detection systems

3. Electronic security locks and proximity device readers

4. Heating, ventilation (supply and exhaust) and air-conditioning (HVAC) systems

5. High-efficiency particulate air (HEPA) filtration systems

6. HEPA decontamination systems

7. HVAC and exhaust air system controls and control interlocks

8. Airtight isolation dampers

9. Laboratory refrigeration systems

10. Boilers and steam systems


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L backflow preventionist of laboratory systemsin the commissioning plan – cont.

11. Fire detection, suppression and alarm systems

12. Domestic water backflow prevention devices

13. Processed water systems (i.e. reverse osmosis, distilled water)

14. Liquid effluent treatment and neutralization systems

15. Plumbing drain primer systems

16. Chemical decontaminant systems

17.Medical laboratory gas systems

18. Breathing air systems

19. Service and instrument air systems

20. Cascading pressure differential verification of laboratories and support areas

21. Local area network (LAN) and computer data systems


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L backflow preventionist of laboratory systemsin the commissioning plan – cont.

22. Normal power systems

23. Emergency power systems

24. Uninterruptible power systems

25. Emergency lighting systems

26. Lighting fixture penetration seals

27. Electrical and mechanical penetration seals

28. Telephone systems

29. Airlock door control interlocks

30. Airtight door seals

31.Window and vision-panel penetration seals

32. Barrier pass-through penetration


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L backflow preventionist of laboratory systemsin the commissioning plan – cont.

33. Structural integrity verification: concrete floors, walls and ceilings

34. Barrier coating verification: floors, walls and ceilings

35. Biosafety Level 4 containment envelope pressurization and isolation functions

36. Biological safety cabinets

37. Autoclaves

38. Liquid nitrogen system and alarms

39.Water detection systems (e.g. in case of flooding inside containment zone)

40. Decontamination shower and chemical additive systems

41. Cage-wash and neutralization systems

42.Waste management.


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Failures in BSL-laboratories backflow prevention

  • August 2007 FMD outbreak at Pirbright in Surrey, UK

    • 3rd August: first case of FMD fount at a farm in Normandy

    • 6th August: second case of FMD at a farm near the first

    • 6th August: FMD strain identified as O1 BFS67

      • Strain not currently found naturally in the world

      • Strain originates from the 1967 FMD epidemic in the UK

      • Strain used as reference in laboratories and pharmaceutical production plants


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Turkish AI Laboratories backflow prevention

  • Eight regional Veterinary Control and Research Institutes (VCRIs) provide laboratory services with the ability to achieve virus isolation. Three of them, Bornova, Pendik and Etlik house AI laboratories in charge of virus identification. Bornova houses the national reference laboratory. They areequipped and competent to undertake a broad range of standard diagnostic HPAI tests and provide technical backstopping to the other 5 regional laboratories authorized for AI diagnostics. However the conditions of those three laboratories do not meet the required bio-safety standards. Therefore GDPC plans to upgrade them to BSL 3.


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UK FMD outbreak in 2007 backflow prevention

  • Pirbright

    • Area holds two BLS laboratories

      for work with FMD

    • Labs used by three organizations:

      • Institute for Animal Health (IAH)

      • Merial Ltd

      • Stabilitech Ltd.

    • IAH and Stabilitech used onlysmall amounts of live FMD.

    • Merial produced large volumes of FMD vaccine.


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UK FMD outbreak in 2007 backflow prevention

  • Findings of the investigation:- Containment failure due to:

    • Inefficient inactivation of waste water (biosafety breach)

    • Broken waste water piping due to poormaintenance

    • High precipitation allowed sewers to overflow

    • Construction work at the site allowed entry and exit of unsupervised personnel and vehicles (biosecurity breach)

      Source: Final report on potential breaches of biosecurity at the Pirbright site 2007. September 2007, available at:

      http://www.hse.gov.uk


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Other recent failures backflow prevention

  • Texas A&M University (2006 – 2007)

  • New BSL-4 laboratory at the CDC (2007)

  • Lessons learned:

    • All possible contingencies must be in place to ensure containment and security

    • Incidents must be reported

    • Establishment of BSL-3 or BSL-4 laboratory is a long term financial commitment, not just an initial one

      Source: High-Containment Biosafety Laboratories: Preliminary Observations on the Oversight of the Proliferation of BSL-3 and BSL-4 Laboratories in the United States. October 4, 2007, published by the GAO, at p. 14.


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