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

2. Contents 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.
This presentation provides a step-by-step approach to the design, construction, and commissioning of animal and human health laboratories.
It will cover:
Brief comment of issues related to national assessment for laboratory needs
Determination of functions and scope of work in a laboratory – i.e. disease and expected test capabilities and expected volumes of each
Resources required – equipment, staff, reagents, ancillary services, etc.
Approach to design with checklists and pitfalls – technical, environmental, safety and perhaps comment on implications for building renovations.
Biosafety considerations
Construction – tips and resources
Commissioning the facility

3. Laboratory biosafety & biosecurity - definitions
“Laboratory biosafety” is the term used to describe the containment principles, technologies and practices that are implemented to prevent unintentional exposure to pathogens and toxins, or their accidental release.
“Laboratory biosecurity” refers to institutional and personal security measures designed to prevent the loss, theft, misuse, diversion or intentional release of pathogens and toxins.
Biosafety
The application of combinations of laboratory practice and procedure, laboratory facilities, and safety equipment when working with potentially infectious microorganisms.
BIOSECURITY
Today, the world is facing a new challenge in safeguarding the public health from potential terrorism involving the use of dangerous biological agents or toxins. Existing standards and practices may require adaptation to ensure protection from such hostile actions. Therefore the prospect of bioterrorism gives rise to the need to protect facilities that work store or transfer dangerous biological materials from being intentionally misused for malevolent ends.

4. Laboratory biosecurity as a complement to laboratory biosafety
Laboratory biosafety and biosecurity mitigate different risks, but they share a common goal: keeping valuable biological materials safely and securely inside the areas where they are used and stored.
Good laboratory biosafety practices reinforce and strengthen laboratory biosecurity systems.

5. Conflicts (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
Even though biosafety and laboratory biosecurity are in most respects compatible, a number of potential conflicts exist that need to be resolved e.g.:
Controls that reduce unauthorized access might also hinder an emergency response by fire or rescue personnel. Therefore mechanisms need to be established that allow entry by emergency responders but ensure uninterrupted and constant laboratory biosecurity, control, accountability and traceability. Also, staff members must be able to quickly and safely exit a laboratory during an emergency without at the same time allowing unrestricted access to the laboratory premises equipment and materials.
Supervised by appointed Biosafety officer
Regulated by national work environment safety law
Incident reporting
Incident response planning
Revision
Training
Supervised by appointed Biosecurity or Biosafety officer
Should consult with law enforcement officials and security experts

6. Why biosafety practices?
Protection:
- workers
- “products”
- co-workers
- lab support personnel
- environment

7. Biosafety levels
BSL1 - agents not known to cause disease (no or low individual and community risk).
BSL2 - agents that cause human or animal diseases with moderate individual or low community risk (e.g. blood borne diseases).
BSL3 - indigenous/exotic agents associated with human disease and with potential for aerosol transmission - high individual risk (respiratory) low community risk)
BSL4 - dangerous/exotic agents of life threatening nature – serious diseases readily transmitted.
Risk Group 1 (no or low individual and community risk) - A microorganism that is unlikely to cause human or animal disease.
Risk Group 2 (moderate individual risk, low community risk) - 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 environment. Laboratory exposures may cause serious infection, but effective treatment and preventive measures are available and the risk of spread of infection is limited.
Risk Group 3 (high individual risk, low community risk) - A pathogen that usually causes serious or potentially lethal human or animal disease as a result of exposure by the inhalation route, but does not ordinarily spread from one infected individual to another. Effective treatment and preventive measures are available.
Risk Group 4 (high individual and community risk) - A pathogen that usually causes serious human or animal disease and that can be readily
transmitted from one individual to another, directly or indirectly. Effective treatment and preventive measures are not usually available.
Biosafety level designations are based on a composite of the design features, construction, containment facilities, equipment, practices and operational procedures required for working with agents from the various risk groups.
Diagnostic and health-care laboratories (public health, clinical or hospital-based) must all be designed for Biosafety Level 2 or above.

8. Biosafety Level 3 Risk 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?
Medical surveillance criteria is based on a risk assessment of the biological agents used.
The assignment of an agent to a biosafety level for laboratory work must be based on a risk assessment. Such an assessment will take the risk group as well as other factors into consideration in establishing the appropriate biosafety level. For example, an agent that is assigned to Risk Group 2 may generally require Biosafety Level 2 facilities, equipment, practices and procedures for safe conduct of work. However, if particular experiments require the generation of high-concentration aerosols, then Biosafety Level 3 may be more appropriate to provide the necessary degree of safety, since it ensures superior containment of aerosols in the laboratory workplace.
Countries should draw up a national classification of microorganisms, by risk group, taking into account: Pathogenicity of the organism.
2. Mode of transmission and host range of the organism. These may be influenced by existing levels of immunity in the local population, density and movement of the host population, presence of appropriate vectors, and standards of environmental hygiene.
3. Local availability of effective preventive measures. These may include: prophylaxis by immunization or administration of antisera (passive immunization); sanitary measures, e.g. food and water hygiene; control of animal reservoirs or arthropod vectors.
4. Local availability of effective treatment. This includes passive immunization, postexposure vaccination and use of antimicrobials, antivirals and chemotherapeutic agents, and should take into consideration the possibility of the emergence of drug-resistant strains.

9. Biosafety Level 3 Risk 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?

10. Biosafety Level 3 Risk 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?
When dealing with an unknown agent……
Is there any known epidemiological data?
Are there any patterns parallel to other agents?
Is there any data from animal studies?
Is the route of infection known?

11. Relation of risk groups to biosafety levels, practices and equipment
Biosafety level (BSL)
Laboratory type
Lab. practice
Safety equipment 1
Basic BSL- 1
Basic teaching
and research
Good microbiol. techniques (GMT)
None, open bench work 2
Basic BSL- 2
Diagnostic services and research
GMT + protective clothing biohazard
sign
Open bench plus bio – safety cabinet (BSC) for potential aerosols 3
Containment
BSL- 3
Special diagnostic services and research
As BSL-2 plus special clothing controlled access directional airflow
Biosafety cabinet and/or other primary devices for all activities 4
Maximum Containment
BSL- 4
Dangerous pathogen units
As BSL-3 plus airlock entry, shower exit and special waist disposal
Class-3 BSC or positive pressure suites in conjunction with class-2 BSCs, double ended autoclave trough the wall and filtered air

12. Summary of biosafety level requirements
Isolation of laboratory No No Yes Yes
Room sealable for decontamination No No Yes Yes
Ventilation:
— inward airflow No Desirable Yes Yes
— controlled ventilating system No Desirable Yes Yes
— HEPA-filtered air exhaust No No Yes/Nob Yes
Double-door entry No No Yes Yes
Airlock No No No Yes
Airlock with shower No No No Yes
Anteroom No No Yes —
Anteroom with shower No No Yes/Noc No
Effluent treatment No No Yes/Noc Yes
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

13. BIOSAFETY AND BIOSECURITY LABORATORY DESIGN CRITERIA
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
The laboratory must be separated from areas which are open to unrestricted traffic flow within the building. Passage through two sets of self-closing doors is the basic requirement for entry into the laboratory from access corridors or other contiguous areas. Clothes changing rooms and shower should be included in the passage way.
Additional separation may be achieved by placing the laboratory at the blind end of a corridor, or constructing a partition and door or access through an anteroom (a specific area designed to maintain the pressure differential between the laboratory and its adjacent space, with a double-door entry). The anteroom should have facilities for separating clean and dirty clothing and a shower may also be necessary. Anteroom doors may be self-closing and interlocking so that only one door is open at a time. A break-through panel may be provided for emergency exit use.
All breaches (holes in wall, cracked or broken ceiling tiles, etc..) must be repaired as soon as possible because they could compromise the bio-containment and affect the air balance.
All surfaces should be of impermeable wipe-clean materials
Ample space must be provided for safe conduct of laboratory work and for cleaning and maintenance.
Walls ceilings and floors should be solid, smooth, easy to clean, impermeable to liquids and resistant to the chemicals and disinfectants normally used in the laboratory. Wood is not appropriate because it absorbs liquid such as blood. Tiles are not appropriate because biological material can seep in between the tiles making decontamination virtually impossible.
The laboratory room must be sealable for decontamination. Air-ducting systems must be constructed to permit gaseous decontamination.
Windows must be closed, sealed and break-resistant.
There must be a controlled ventilation system that maintains a directional airflow into the laboratory room.
The building ventilation system must be so constructed that air from the containment laboratory – Biosafety Level 3 is not recirculated to other areas within the building. Air may be reconditioned and recirculated within that laboratory if it is HEPA filtered. When exhaust air from the laboratory (other than from biological safety cabinets) is discharged to the outside of the building, it must be dispersed away from occupied buildings and air intakes. This air should be discharged through HEPA filters.
A heating, ventilation and air-conditioning (HVAC) control system with visual monitoring device should be installed so that staff can at all times ensure that proper directional airflow into the laboratory room and negative pressure is maintained. Consideration should be given to the installation of audible or clearly visible alarms to notify personnel of HVAC system failure.
Laboratory furniture should be sturdy with smooth surfaces, impervious to water and resistant to acids, alkalis, organic solvents, and moderate heat. Wooden bench tops are not appropriate because an unfinished wood surface can absorb liquids. Fiberglass is inappropriate as well since it can degrade when strong disinfectants are applied.
Vacuum lines are protected with liquid disinfectant traps and HEPA filters, or their equivalent, which are routinely maintained and replaced as needed. All HEPA filters must be installed in a manner that permits gaseous decontamination and testing.
Facilities for storing outer garments and personal items should be provided outside the laboratory working areas.
Facilities for eating and drinking and for rest should be provided outside the laboratory working areas.
Doors should have vision panels, appropriate fire ratings, and preferably be selfclosing.
Safety systems should cover fire, electrical emergencies, emergency shower and eyewash facilities.
First-aid areas or rooms suitably equipped and readily accessible should be available In the planning of new facilities, consideration should be given to the provision of mechanical ventilation systems that provide an inward flow of air without recirculation.
Each laboratory room should contain a sink for handwashing. The sink is foot, elbow, or automatically operated and is located near the laboratory exit door. An eyewash facility must also be readily available.
Biological safety cabinets should be sited away from walking areas and out of crosscurrents from doors and ventilation systems. If class I or class II biological safety cabinets are used, the exhaust air from them which will have been passed through HEPA filters, must be discharged in such a way as to avoid interference with the air balance of the cabinet or the building exhaust system.
An double door autoclave for the decontamination of waste material should be available in the containment laboratory. If infectious waste has to be removed from the containment laboratory for decontamination and disposal, it must be transported in sealed, unbreakable and leakproof containers.
Backflow-precaution devices must be fitted to the water supply. Vacuum lines should be protected with liquid disinfectant traps and HEPA filters, or their equivalent. Alternative vacuum pumps should also be properly protected with traps and filters.
Illumination should be adequate for all activities. Undesirable reflections and glare should be avoided.
Storage space must be adequate to hold supplies for immediate use and thus prevent clutter on bench tops and in aisles. Additional long-term storage space, conveniently located outside the laboratory working areas, should also be provided. Space and facilities should be provided for the safe handling and storage of solvents, radioactive materials, and compressed and liquefied gases.
A dependable supply of good quality water is essential. There should be no crossconnections between sources of laboratory and drinking-water supplies. An antibackflow device should be fitted to protect the public water system.
There should be a reliable and adequate electricity supply and emergency lighting to permit safe exit. A stand-by generator is desirable for the support of essential equipment, such as incubators, biological safety cabinets, freezers, etc., and for the ventilation of animal cages.
Physical and fire security must be considered for the laboratories and animal houses . Strong doors, screened windows and restricted issue of keys are compulsory.

14. Essential Building Principles
Primary containment barrier is the first barrier between agent and man (such as gloves, gowns, masks, biosafety cabinets, respiratory protection etc.)
Secondary containment barrier is the barrier between agents and environment (airtight rooms, air handling and filtration, air locks, showers, laundry, sewage treatment, waste disposal, sterilisers, redundant services 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.
Primary barriers pertain to equipment such as gloves, gowns, masks, biosafety cabinets, respiratory protection, and positive-pressure ventilation suits as well as the use of good laboratory techniques.
Secondary barriers are addressed through facility design with airtight rooms, air handling and filtration, air locks, showers, laundry, sewage treatment, waste disposal, sterilisers, redundant services as well as equipment and material niches.
Tertiary barriers deal with the physical operation with items such as walls, fences, security, quarantine and animal exclusion zones. Due to the varying risk of biological agents, the facilities that handle these agents need to be designed and classified accordingly.

15. Work flow considerations
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).
People that do not belong in containment should be restricted from entering containment areas through the use of various security systems, although these people will no doubt need access to the building. Therefore the building may need a series or layers of control devices for access.
The ability to decontaminate equipment should also be considered for a containment facility. Many types of equipment cannot be autoclaved due to their size or sensitivity to steam decontamination, such as microelectronics. Decontamination chambers are an effective use of space for fogging, vaporisation, or gaseous methods. When planning chambers, the size of the space (volume) for decontamination purposes should accommodate the largest anticipated piece of equipment needed.
There are differences between human health and animal health laboratories. Animal health laboratories have additionally necropsy room and animal facilities which are integrated to the laboratory and need special design.

16. The containment barriers
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.
Compartmentalization of each laboratory unit must be achieved by providing
it with at least:
A. One-hour fire-rated separation from adjacent laboratories or other areas
B. Self-closing fire doors with at least a 20-minute fire rating
C. Class A interior finishes
D. Class I floor finishes
E. Doors to corridors from laboratories that swing in the direction of exit
Provide a minimum of two exits in laboratories larger than 200 square feet, where hazardous materials are used.
Aisles serving a single work area must be a minimum of 36 inches wide. Double aisles must be a minimum of 60 inches wide. Avoid aisles longer than 20 feet. Arrange furniture for easy access to an exit from any point in the laboratory.
Faucets, to which a hose or similar device may be attached, must be providedwith an approved vacuum breaker. Alternately, a special laboratory water supply equipped with an RPZ back flow device to separate it from the potable water maybe provided. If a laboratory water system is provided, all connected outlets must be labeled “Not Potable.”
A safety shower and eyewash must be provided in each lab area equipped with a fume hood. In other laboratories using chemicals, eyewash must be required. If feasible, control the water supply to a temperature between 60 degrees F and 95 degrees F. Refer to Appendix S - Emergency Eyewash and Safety Shower Installation. All rooms where Biosafety Level 2 and above organisms are manipulated require an eyewash and a hand wash sink.
Provide a single shut-off valve for each laboratory in accessible locations or central supply of flammable, combustible or oxidizing gases. Valves must be outside of the areas in which the gases are used. These shut-off valves are in addition to those at the points of supply and use. They may be located adjacent to the corridor exit from the lab or, if security is not a problem, in the corridor.
Storage and supply systems for compressed and liquefied gases mustcomply with requirements of NFPA and ANSI. Consult the following standards:

17. Specimen Reception, Dispatch Area and "Grey Areas"
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.

18. Security and related systems
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
Security solutions for containment facilities need to reflect local, regional, national and international risk analysis that is measured at a point of time and also anticipates future considerations. The present day approach to designing containment facilities need to reflect on recent past terrorism incidents and the increased awareness of the general public to the scientific operations. Threat and Risk Assessments (TRA) are normally done early in project planning to establish the features necessary to secure the facility, assets (which could include animals) and biological agents. It is correct to state that a well-designed facility could be built both safe and secure anywhere regardless of local risks, this however is tempered with the fact that all risks and the consequences of the risk occurring are very rarely known (e.g. effects of hurricane Katrina). Once a risk assessment is complete technical design solutions can be provided to mitigate risk.

19. Security Zone 1: Property Protection Area (uncontrolled)
• 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

20. Security Zone 2: Limited Areas / Non-Containment
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

21. Security Zone 3: Exclusion Area / Non-Containment
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

22. Security Zone 4: Exclusion Areas – Containment
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

23. Security Zone 5: Exclusion Areas – Containment
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

24. 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.
Authorisation versus Authentication
Appropriate security access control strategies ensure the right people have the appropriate access control. There is a difference between authorisation and authentication. Optimum secured access control provides three access control features:
• You have something with you that gives you authorisation (card, key)
• You know something that only you have knowledge of giving you access (PIN, memory)
• You are uniquely identified based on your biometrics (fingerprint, hand geometry, voice, retinal scan, DNA etc.)
Possession of a card or key does not authenticate individual identity. Individual knowledge of a pin reinforces individual identity and supplements card or key access control but is not a foolproof confirmation of identity. Biometric validation technologies provide the most foolproof authentication of individual identity. Containment facilities pose a unique challenge when factoring the three methods of access control. The use of cards gaining access to outside perimeters is acceptable until access control is required inside containment. Per protocol, employees are to shed all clothes and materials and wear new clothes and gloves for specific labs and primary containment zones.
As a general rule, access to secondary barrier zones will be controlled using card access security systems. Access to inside labs or animal rooms will be controlled using either memory, biometric or a combination of both. All security control systems are subject to potential failure scenarios based on False Acceptance Rates (FAR) and False Rejection Rates (FRR). BSL-3 Laboratory SOPs should include personnel security training and operational safety plans in the event of a security breach.

25. Aerosol Control Sources of Biohazardous Aerosols
Impact of Ventilation on Aerosol Load
Air filtration
Airlocks
Anterooms as a Control Mechanism
Cascade negative pressure
SOP and PPE as a Control Mechanism
There are many examples of airlocks, but effective airlocks 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

26. Sources of Biohazardous Aerosols
Biohazardous aerosols of concern in the laboratory setting are generated by a number of manipulations involving infectious material. such as sonification, 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.

27. Ventilation
Issues related to ventilation in containment facilities include: directional airflow, airflow velocities, pressure differential between adjacent spaces and air exchange rates.
Directional airflow is used to create zones of hazard by moving air from areas clear of hazardous aerosol contamination to areas of higher potential for hazardous aerosol contamination. This provides for two functions:
- 1) control of the hazardous aerosol minimises the possibility of inadvertent exposure outside of the laboratory space and;
- 2) knowledge of where the aerosol hazard exists and the extent of the hazard allows personnel to follow appropriate protocols if they are required to enter areas where aerosols may exist.
It should be noted that aerosol control by directional airflow is generally effective from one space separated by a door to another space. The ventilation system must not circulate exhaust air.

28. Air filtration
Where the risk assessment indicates that a significant aerosol release of pathogens outside of primary containment is probable and would create a hazard to people or the environment outside of the facility, the exhaust system should be HEPA filtered to prevent the release of the pathogens outside of the laboratory.

29. Airlocks
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 (a device 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.

30. Airlocks- cont. 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

31. Anterooms and two doors in series
Considerable control of airborne micro-organisms can be achieved with the addition of an anteroom to the laboratory or animal holding room.
This is the basis for the requirement 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.

32. Cascade negative pressure
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.

33. Technical details about pressure differentiation and backflow prevention
Pressure differentials in animal facilities are held at approximately 50 Pa lower pressure than the point of personnel 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.

34. Electrical 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.
Water and electricity should be kept apart whenever possible. Electrical systems need to be protected. Rodents and vermin can also create havoc by chewing through wire insulating jackets and nesting inside equipment causing circuit disruption, fires and dust.

35. Emergency 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
These systems should have UPS (Uninterruptible Power Supply) on the control systems associated with the management of data, fail-safe positions, monitoring and alarming UPS systems can be installed per system, on a zone basis or on a central basis. An analysis on costs for determining which one is preferable should be done in advance of design.
If possible refrigerators and freezers should be housed in dedicated rooms or areas; this allows a panel board with a time delay on energisation to be installed to prevent the immediate restart of the compressors after a power bump. This will save unnecessary wear and tear on the compressors. This will also allow the additional cooling required to be provided to the dedicated refrigerator / freezer room

36. Emergency Power
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.

37. Identification
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.

38. The 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 and circuit 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)

39. Labels
The labels should be colour coded to provide indication of the system. For example:
• 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.

40. Indicator Lights
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.

41. Effluent treatment
Heat treatment – 95 C
Chemical treatment

42. Redundancy
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.

43. Laboratory animal facilities
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.

44. Planning Experimental Work
In an animal bio-containment facility, a basic assumption is that animals will not bring any disease into the facility that will compromise either the planned experiment or other animals. Therefore animals must be introduced via a path that is free from disease agents and that no disease agent will escape from within while fresh animals are introduced.
Access to animal rooms is limited to personnel that have been advised of potential hazards, are trained, meet specific requirements.
In some jurisdictions the animal welfare legislation requires that staff handling the animals must be trained in each species they handle or do any procedures on. In general, persons who are at increased risk of acquiring infection, or for whom infection might be unusually hazardous, are not allowed into the animal room.

45. Waste Disposal
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 be decontaminated either by heat or chemical treatment.
Discarded surgery or necropsy tissues from infected animals are usually sterilised by autoclaving and carcasses by rendering at high temperature, steam sterilisation, incineration or chemical decontamination such as alkaline hydrolysis.
All infectious wastes that cannot be decontaminated or autoclaved will immediately be placed in red infectious waste bags.
All infectious / objectionable (other than sharps and research animal waste) intended for onsite decontamination by autoclaving will be placed immediately into approved, prominently labelled, leak-burst-tear, and puncture proof containers at the point of generation. Infected animal carcasses and body parts may be decontaminated on-site or shipped in appropriate transport containers for off-site incineration.
All infectious wastes that cannot be decontaminated or autoclaved will immediately be placed in red infectious waste bags. When the bag is 3/4 full, or at the end of experiment or procedure – depending on the infectious agent – the bag will be sealed, placed in a disposal container and stored in freezers, refrigerators, or cold rooms until shipment to outside disposal facility.
Use of sharp instruments should be restricted whenever possible. All used disposable sharps will be placed directly into approved prominently labelled, leak and puncture-resistant prominently labelled disposable sharps containers.

46. Laboratory commissioning
Laboratory commissioning may be defined as the systematic review and documentation process signifying that specified laboratory structural components, systems and/or system components have been installed, inspected, functionally tested and 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.
Commissioning involves testing and validating the performance of the laboratory components and integrated systems.
Containment laboratories require special attention to normal and critical systems to ensure their operating performances both in normal and emergency situations do not compromise protection for the worker, environment or the program.
Commissioning of laboratory systems are necessary to prove to regulatory authorities that the facility has taken the necessary precautions to work with hazardous and infectious agents.
Building systems commissioning is a process designed to ensure that the finished facility, equipment and systems will operate in accordance with the design intent and construction documents. It is recommended that commissioning be implemented early in the planning phase through to the construction and certification. To ensure that the physical requirements for the intended containment level and use of the facility have been met, each laboratory must undergo a detailed commissioning treatment. This requires verification and documentation of critical containment components, equipment start-up, control system calibration, balancing and performance testing. A complete set of drawings and specifications, an understanding of the intended use and work to be performed, a list of equipment requirements, all test results, and an understanding of the intent of the systems’ operation are all part of the commissioning process. Containment laboratory projects are unique in that the systems, equipment, architecture, protocols and researchers work together cohesively have to ensure biocontainment, biosafety and biosecurity.

47. Why laboratory commissioning?
The commissioning process provides the institution and the surrounding community with a greater degree of confidence that the structural, electrical, mechanical and plumbing systems, containment and decontamination systems, and security and alarm systems will operate as designed, to assure containment of any potentially dangerous microorganisms being worked with in a particular laboratory or animal facility.

48. List of laboratory systems in the commissioning plan
1. Building automation systems including links to remote monitoring and control sites
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

49. List of laboratory systems in 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

50. List of laboratory systems in 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

51. List 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.

52. Failures in BSL-laboratories
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

53. Turkish AI Laboratories
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 are equipped 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.
There are no clearly detached clean and durty zones and physical barriers between them. There are no changing rooms or shower facilities at the entrance. There are no decontaminatretment facilities for heat or chemical treatment of the waste water or liquides. There are no facilities for decontamination of infected solid materials from the laboratory like duble ended door autoclave or disinfection fumigation chamber. There are no ventilation systems equipped with HEPA filters and working under negative preasure to protect airosol spread of the virus.

54. UK FMD outbreak in 2007 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 only small amounts of live FMD.
Merial produced large volumes of FMD vaccine.

55. UK FMD outbreak in 2007
Findings of the investigation: - Containment failure due to:
Inefficient inactivation of waste water (biosafety breach)
Broken waste water piping due to poor maintenance
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 September 2007, available at:

56. Other recent failures 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.