Containment Standards for Veterinary Facilities
Sections 6-8

6. Operational Practices

6.1 General Requirements

The following general practices are required when working in any containment laboratory or animal facility:

entry must be restricted to laboratory staff, animal handlers, maintenance staff and other persons on official business
only persons meeting specific entry requirements (e.g. immunization, serum screening) may enter containment laboratories unless the facility has been appropriately decontaminated
a health and medical surveillance program must be provided as recommended by Health Canada
personnel must receive training on the potential hazards associated with the work involved and the necessary precautions to prevent exposures to zoonotic agents and release of non-indigenous agents; personnel must show evidence that they understood the training provided; training must be documented and signed by both the employee and supervisor
a documented procedural manual must be written and followed
all persons (including visitors, maintenance staff, etc.) entering the containment area must be trained and know and follow the operational protocols for the project in process; trainees must be accompanied by a trained staff member
persons entering a containment facility must be well prepared and bring all materials they will need with them; if something has been forgotten, traffic patterns must still be adhered to (ie. do not go back to get it; either phone for someone to bring it or exit via proper protocols)
employees working in the containment area must have general knowledge of the physical operation and design of the facility (e.g. air pressure gradients between zones, directional air flow patterns, alarm signals for air pressure failure, containment perimeter)
traffic flow patterns from clean to dirty areas must be established and adhered to (ie. move from least to most contaminated areas)
smoke testing (ie. with a smoke pencil) should be done periodically by lab staff to verify correct airflow
entry/exit protocols for persons, animals, equipment, samples, waste, etc. must be written, posted and followed; general protocols must be supplemented with protocols specific for each project in progress
emergency procedures for entry/exit, spill clean-up, air handling/biosafety cabinet failure, fire, animal escape and other emergencies must be written, posted and followed
in the event of life-threatening emergencies, personal health and safety are a priority; exit protocols must be established whereby routine procedures are bypassed; a reporting area must be identified where further steps must be taken (e.g. disinfecting footwear, changing, showering) prior to leaving
all spills, accidents, overt or potential exposures to infectious materials, and losses of containment (e.g. lab positive pressurization) must be reported immediately to the laboratory supervisor; written records of such incidents must be maintained
an effective rodent and insect control program must maintained

6.2 Laboratories

The following describes the minimum operational practices required at AP containment level 2:

laboratory personnel must be trained in and follow the safe use of laboratory equipment, biological safety cabinets, procedures to minimize the production of aerosols, decontamination and emergency response
open wounds, cuts, scratches and grazes should be covered with waterproof dressings
eating, chewing gum, drinking, smoking, storing food, and applying cosmetics are prohibited
personal items such as purses and outdoor clothing should be kept separate from work areas
the work area containing hazardous materials should be kept free from materials not pertinent to the work and that cannot be easily decontaminated (e.g. journals, books, correspondence); paperwork and report writing should be kept separate from such work areas
laboratory reference material should be kept in the laboratory zone
hands should be washed frequently (after handling infectious materials, after removing gloves, and before leaving the laboratory)
open-toed and high-heeled shoes must not be worn in the laboratory
long hair should be tied back so that it cannot come into contact with hands, specimens, containers, or equipment
gloves (e.g. intact vinyl or latex) must be worn when handling infectious materials; metal mesh gloves can be worn underneath the latex or vinyl glove to provide protection from sharps and needles
laboratory coats, gowns or coveralls must be worn when working in the laboratory; this clothing must not be worn in non-laboratory areas (e.g. offices, staff rooms, canteens, libraries)
protective lab clothing should not be stored in the same locker as street clothing
contaminated clothing must be decontaminated prior to laundering (unless laundering facilities are within the laboratory zone and have been proven to be effective in decontamination)
eye and face protection must be worn when it is necessary to guard against splashing hazardous materials, flying particles, and harmful light or other rays
laboratory doors must be kept closed as required by the facility design
biological safety cabinets must be used for procedures with potential for producing infectious aerosols (e.g. with zoonotic agents) and with high concentrations or large volumes of zoonotic materials
contaminated work surfaces must be decontaminated
all contaminated materials must be decontaminated before disposal or cleaning for reuse
contaminated equipment leaving the laboratory for servicing or disposal must be appropriately decontaminated
efficacy monitoring of autoclaves using biological indicators must be done at least weekly, depending on the frequency of use of the autoclave, and records of the results kept on file; cycle log records (ie. time, temperature and pressure) must also be kept on file

In addition to the general operational practices listed for level 2, the following describes the minimum operational practices required at AP containment level 3:

a protocol specific to the operation of the lab must be developed and read by personnel; employees must certify in writing that they have understood the material in the protocol
the laboratory zone must be kept locked
infectious agents should be stored inside the laboratory zone; agents stored outside the zone must be kept locked, in leakproof containers
personnel must have demonstrated proficiency in microbiological practices and techniques (e.g. experience in handling infectious organisms or cell cultures)
personal items such as purses and outdoor clothing must not be brought into the laboratory zone
a containment check must be performed prior to entering the laboratory zone (ie. verify negative lab pressurization as designed)
water seals must be maintained in drainage traps (ie. through regular sink/shower usage and/or by filling traps in areas that are not being used)
laboratory samples and supplies may be carried into the laboratory zone or passed through a ventilated pass-box; where the barrier autoclave is used to pass materials into the laboratory, the autoclave must have been cycled prior to opening the outer "clean side" door
personnel entering the laboratory zone must remove street clothing and jewellery, and change into dedicated laboratory clothing and shoes
where full body protective clothing is not worn a shower is required on exit from the laboratory; where a known or suspected aerosol exposure has occurred (e.g. dropping infectious materials) a shower is required on exit from the laboratory zone
a shower (including washing hair, beards) is required on exit from a laboratory zone handling non-indigenous animal pathogens; eye glasses must be disinfected at the containment barrier
a second layer of protective clothing (ie. solid-front gowns with tight-fitting wrists, gloves) should be worn over laboratory clothing when directly handling infectious materials (e.g. dedicated for use at the biological safety cabinet)
contaminated clothing must be decontaminated prior to laundering (unless laundering facilities are within the laboratory zone and have been proven to be effective in decontamination of the microorganisms likely to be encountered)
all activities with infectious materials are conducted in a biological safety cabinet; where this is not possible, other physical containment devices in combination with personal protective clothing and equipment must be used; no work with open vessels containing infectious materials is conducted on the open bench
centrifugation of infectious materials must carried out in sealed safety cups or rotors that are loaded and unloaded in a biological safety cabinet
all contaminated waste materials leaving the laboratory zone must be decontaminated through a double-door autoclave at the barrier before disposal; both doors of the autoclave must not be opened simultaneously
heat sensitive materials that cannot be autoclaved out of the laboratory zone must be decontaminated at the containment barrier (e.g. fumigated with formaldehyde or vaporized hydrogen peroxide, disinfected using liquid chemicals, or other technology proven to be effective)

In addition to the general operational practices listed for level 2 and 3, the following describes the minimum operational practices required at AP containment level 4:

all persons entering the laboratory zone must have completed a training course in procedures specific to the level 4 lab and must show evidence of having understood the training; training must be documented and signed by the employee and supervisor
protocols must be established for emergencies including personnel suit damage, loss of breathing air, and loss of chemical shower
where HC level 4 agents are used, employees must carry an illness surveillance card (e.g. with employee name, supervisor's and alternate's name and phone number, facility phone number); employees must immediately notify their supervisor of any febrile illness; supervisors must contact any employee with unexplained work absences
a log record is maintained of activities in the laboratory
infectious agents must be stored inside the laboratory zone
a daily check of lab systems must be carried out
personnel entering the laboratory must remove street clothing and jewellery, and change into dedicated laboratory clothing and shoes
when HC containment level 4 zoonotic agents are manipulated, positive-pressure suits must be worn; suit integrity must be routinely checked for leaks
a shower is required on exit from the laboratory zone
a chemical shower of appropriate duration is required when suits are worn; the disinfectant used must be effective against the agents of concern, be diluted as specified and prepared fresh as required

6.3 Animal Facilities

Work with animals poses a variety of special hazards including exposure to zoonotic agents (naturally occurring or experimentally infected), animal bites and scratches, kicks and crushing injuries, physical hazards (e.g. noise, temperature) and chemical hazards (e.g. cleaning agents, disinfectants). Allergic conditions can result from contact with animal fur or hair, bedding, and animal wastes. At least one-fifth of people who work with laboratory rodents, guinea pigs and rabbits develop allergies. Protection from allergens must be provided through engineering controls, ventilation, use of isolators and cages with filter tops and appropriate use of respiratory protection.

Animal handlers must have knowledge of the species' general characteristics such as behaviour, instincts and physical attributes. Consideration should also be given to their natural ecto- and endoparasites and the zoonotic diseases to which they are susceptible including their route of excretion and dissemination.

The following describes the minimum operational practices required in small animal facilities (SA facilities):

coveralls and footwear must be worn when working in AP containment level 2 SA facilities
personnel entering AP containment level 3 and 4 SA facilities must change into dedicated facility clothing and shoes
gloves must be worn when handling infected animals
hands must be washed after handling animals, after removing gloves and before leaving the facility
HEPA-filtered respirators are required for handling animals in AP containment level 2 and 3 SA facilities where infectious aerosols of zoonotic agents may be generated and cannot be contained within a primary containment device
positive pressure ventilated suits are required for handling animals infected with HC containment level 4 zoonotic agents
a shower (including washing hair, beards) is required on exit from AP containment level 3 SA facilities handling non-indigenous agents and when handling other agents where aerosols cannot be contained in primary containment devices; eye glasses must be disinfected at the containment barrier
a shower is required on exit from AP containment level 4 SA facilities; a chemical shower is required when suits are worn
where a clothing change is not performed on exit from each animal room, disinfectant footbaths are required (the disinfectant must be effective against the microorganisms of concern and changed regularly in accordance with the active life of the disinfectant)
contaminated clothing must be decontaminated prior to laundering (unless laundering facilities are within the laboratory zone and have been proven to be effective against the microorganisms likely to be encountered)
each animal room must be labelled with unique hazards and entry requirements (e.g. respiratory protection)
animal room doors must be kept closed as required by facility design
water seals must be maintained in drainage traps (ie. through regular sink/shower/floor drain usage and/or by filling traps in areas that are not being used)
cages housing infected animals must be appropriately labelled
containment caging systems should be used in AP containment level 3 SA facilities to contain aerosols (e.g. laminar flow cabinets, solid wall and bottom cages covered with filter bonnets)
containment caging systems must be used in AP containment level 4 SA facilities
careful handling procedures must be employed so as to minimize the creation of aerosols and dissemination of dust from cages, refuse and animals
proper methods of restraint must be used to minimize scratches, bites and accidental self-inoculations
necropsy of small animals infected with a zoonotic agent and intranasal inoculation of animals in AP containment level 3 and 4 SA facilities should be conducted in a biological safety cabinet; animals must be securely transported to the biological safety cabinet
supplies and materials are brought into the AP containment level 3 and 4 SA facilities by carrying them in or by way of a ventilated pass-through; barrier autoclaves, fumigation chambers or airlocks may also be used providing that they have been decontaminated before opening the outer "clean side" door
animal bedding must be removed in a manner that minimizes the generation of aerosols and dust; for AP containment level 3 and 4 SA facilities handling zoonotic agents, cages must be decontaminated prior to removing bedding
all contaminated materials must be decontaminated before disposal or cleaning for reuse
autoclaving is the preferred method of decontaminating cages prior to washing; temperature of the final rinse water in mechanical cage washers should be at least 82ºC
animal carcasses and tissues must be incinerated or processed through new technology proven to be effective (e.g. tissue autoclave); carcasses must be transported from the animal room for disposal in leakproof containers that are appropriately labelled

The following describes the minimum operational practices required in large animal facilities (LA facilities):

coveralls and footwear must be worn when working in AP containment level 2 LA facilities
personnel entering AP containment level 3 and 4 LA facilities must remove street clothing and jewellery, and change into dedicated facility clothing and shoes
personnel entering AP containment level 3 animal cubicles where non-indigenous agents are used, level 3 animal cubicles with a single corridor design and level 4 animal cubicles must change into dedicated cubicle clothing and boots at the cubicle entrance
where a clothing change is not performed on entry/exit to the animal cubicle, a second layer of protective clothing must be donned when entering the animal cubicle (e.g. rubber boots, gloves, rubber suit); this layer must be decontaminated on exit from the animal cubicle (e.g. disinfectant foot baths)
gloves must be worn when handling infected animals
hands must be washed after handling animals, after removing gloves and before leaving the facility
HEPA-filtered respirators are required for handling animals where infectious aerosols of zoonotic agents may be generated
positive-pressure ventilated suits are required for handling animals infected with HC containment level 4 zoonotic agents
a shower (including washing hair, beards) is required on exit from AP containment level 3 LA and level 4 LA facilities; eye glasses must be disinfected at the containment barrier
a shower is required on exit from AP containment level 3 animal cubicles where non-indigenous agents are used, level 3 animal cubicles with a single corridor design, and level 4 animal cubicles; eye glasses must be disinfected at the containment barrier
contaminated clothing must be decontaminated prior to laundering (unless laundering facilities are within the laboratory zone and have been proven to be effective against the microorganisms likely to be encountered) (note: autoclaving heavily soiled laundry (e.g. blood, feces) may cause stains to "lock in"; in such cases it may be necessary to launder the clothing first, providing the laundry machines are located within containment)
each animal cubicle must be labelled with unique hazards and entry requirements (e.g. respiratory protection)
animal cubicle doors must be kept closed as required by facility design
water seals must be maintained in drainage traps (ie. through regular sink/shower/floor drain usage and/or by filling traps in areas that are not being used)
proper methods of restraint must be used to minimize kicks, crushing injuries and accidental self-inoculations
supplies and materials are brought into the AP containment level 3 and 4 LA facilities by carrying them in or by way of a ventilated pass-through; barrier autoclaves, fumigation chambers or airlocks may also be used providing that they have been decontaminated before opening the outer "clean side" door
animal are brought into the AP containment level 2,3, and 4 LA facility by means of an airlock
entering more than one animal cubicle from a clean corridor is generally not acceptable; entering more than one animal cubicle from the dirty corridor may be acceptable depending on the project (e.g. moving between contaminated areas of equal status; working with negative control animals first before proceeding to infected animals)
the exterior surfaces of containers of biological samples to be removed from contaminated animal rooms must be decontaminated; heat sensitive samples can be chemically disinfected (e.g. immersion in disinfectant on the barrier)
at the end of the experiment all supplies remaining in the animal cubicles (e.g. supplies, feed) must be removed and decontaminated
animal carcasses and tissues must be incinerated or processed through new technology proven to be effective (e.g. tissue autoclave); carcasses must be transported from the animal cubicle for disposal via the dirty corridor (alternatively, leakproof containers may be used for transport)
animal cubicles and the dirty corridor must be cleaned and decontaminated at the end of an experiment using an appropriate procedure; the disinfectant must be effective against the microorganisms of concern; preliminary washing using a general purpose disinfectant/detergent should be done using low-pressure hoses; decontamination can then be achieved by spraying or fumigating with a disinfectant as appropriate

6.4 Post-Mortem Rooms

Hazards in the post-mortem (PM) room are not limited to splashes and aerosols of infectious materials. Accidents can be caused by cutting instruments, sharp ends of cracked bones, slippery floors, electrical equipment, chemical fixatives and disinfectants.

General precautions:

only authorized staff are allowed to use the necropsy facilities
staff must be trained in the use of all equipment and tools (e.g. electric hoist/monorail, tools, PM table, incinerator)
staff must be trained in proper disinfection and cleaning procedures
the area must be kept neat and tidy; equipment, paper, reports, etc. should be stored securely and not be accumulated in the PM room to facilitate cleaning and decontamination; floors should be clear of obstructions
specific protocols for each project must be developed and followed; these include entry/exit protocols (for people, animals, equipment and samples), protective clothing and equipment, disinfection and cleaning protocols, use of the incinerator and autoclaves, and emergency procedures

Preparation for necropsy:

protective clothing appropriate to the AP containment level and potential hazards must be worn in the PM room; this should include the removal of street clothing and donning of protective clothing and footwear; HEPA-filtered respirators are required when the potential for infectious aerosols exist; waterproof aprons, gloves and eye/face protection (face shield, goggles) should also be worn; a safety helmet is required when operating an electrical hoist/monorail
specific protocols must be developed for the movement of animals and carcasses into the PM room (e.g. hoist for large animals, cart for small livestock, secure containers for poultry and laboratory animals)

Necropsy Procedures:

necropsy safety procedures specific to the species involved must be followed (ie. use of cutting instruments to avoid injury)
the animals (especially birds and small lab animals) should be wetted with water and/or disinfectant prior to necropsy
skilful technique is required to prevent excessive spread of contamination and the formation of aerosols originating from fluids and tissues (this is particularly important for work with zoonotic agents); every effort should be made to confine the spread of contamination; this is especially true when there is a likelihood of material being dropped from an elevated position

Cleaning and disposal procedures:

upon completion of the post mortem, all necropsy tools and instruments must be decontaminated by autoclaving or disinfection (the disinfectant must be effective against the microorganisms of concern); as some disinfectants are inactivated in the presence of organic materials, gross contamination should be removed prior to disinfection
disposable sharps, needles, blades, glass slides, etc. must be discarded into an appropriate sharps container for decontamination
the necropsy table, floor and other contaminated work areas must be cleaned and disinfected at the end of an experiment using an appropriate procedure; preliminary washing using a general purpose disinfectant/detergent should be done; special care must be exercised when using a hose to wash the area (ie. prevent the spread of contamination and formation of aerosols); decontamination of the PM room can then be achieved by spraying or fumigating with a disinfectant effective against the microorganisms of concern
specimens (fresh, frozen or fixed) for further study should be placed in leakproof containers, appropriately labelled; the outside of the container must be cleaned and disinfected at end of necropsy or upon exit from the PM room; samples may only be opened in a laboratory zone of the same AP containment level
all animal waste must be incinerated or processed through new technology proven to be effective (e.g. tissue autoclave); the incinerator or tissue autoclave should be located adjacent to the PM room
where large specimens must be divided into smaller pieces and transported to the incinerator, pieces should be placed carefully into leakproof containers to avoid splashes and aerosols; the outsides of containers must be cleaned down and disinfected prior to transport
out of the PM room; the containers must be labelled with the contents and the name and phone number of a contact person

Exit procedures:

the requirement for showering out of the PM room is dependent on the microorganism of concern; a full shower out of the facility (including washing hair, beards and glasses) is mandatory when working with zoonotic level 3 and 4 agents, and non-indigenous agents
contaminated protective clothing must be decontaminated prior to disposal or re-use; contaminated laundry is autoclaved prior to processing (unless using a pass-through laundry machines proven to be effective against the microorganism of concern)

6.5 Decontamination Procedures

Decontamination is defined as the removal of contamination and includes both sterilization (the complete destruction of all microorganisms) and disinfection (the destruction of specific types of microorganisms). Decontamination procedures for waste disposal, for removing materials, equipment, samples from containment zones, for laundry, for contaminated surfaces and rooms, etc. represent a critical containment barrier. Failure in the procedure can result in the unintentional release of agents from the containment facility. It is the responsibility of each facility to see that proper procedures are followed and that containment is not breached. The choice of method is determined by the nature of the material to be treated.

all decontamination and waste management procedures must be in accordance with applicable federal, provincial, and municipal regulations
all employees must be trained in decontamination procedures specific to their activities and know the factors affecting the effectiveness of the treatment procedure
written procedures must be available for each specific decontamination method being used
all records of efficacy testing and logs of decontamination cycles must be kept on file and be available for inspection as necessary

Several treatment options are briefly discussed below:

Autoclaves:

The effectiveness of decontamination by steam autoclaving is dependent upon the temperature to which the material is subjected as well as the length of time it is exposed. Particular attention must be given to packaging including the size of containers and their distribution in the autoclave. Containers must have good steam permeability and must be arranged in the autoclave in a manner that permits free circulation of steam. Tight fitting containers do not permit steam penetration. Piling containers above one another and overloading can result in decontamination failure.

Effective operating parameters for autoclaves should be established by developing standard loads and their processing times through the use of themocouples and biological indicators placed at the centre of the load. Biological indicators are also used on a regular basis (e.g. weekly, based on the frequency of use) to monitor the effectiveness of the autoclaving cycle. Records must also be kept of the time, temperature and pressure for each load.

Note: Chemical indicators for steam, time and temperature are useful for day-to-day monitoring that the load has been processed, however, must not be used as an indicator of sterility.

Chemical disinfection and fumigation:

Chemical disinfectants are used for the decontamination of surfaces and equipment which cannot be autoclaved; of specimen containers removed from containment; of spills of infectious materials; of protective equipment and clothing; of laboratories, animal cubicles and other rooms; and a variety of other decontamination procedures where heat treatment is not feasible. The choice of chemical disinfectant is dependent on the resistance of the microorganisms of concern (e.g. resistant mycobacteria vs susceptible bacterial species), the application (e.g. liquid vs gaseous fumigation) and the nature of the material to be disinfected (hard surfaces vs porous materials).

The effectiveness of the disinfection procedure can be influenced by the presence of organic material, temperature, relative humidity, concentration of the disinfectant, and contact time. Each of these parameters must be carefully evaluated and defined in accordance with the properties of the disinfectant product and specific disinfection procedure.

Liquid effluent treatment systems:

Liquid effluent treatment systems are typically heat-based and are used for decontaminating liquid waste streams from building sources including sinks, showers, water closets, autoclaves, washing machines and floor drains. The decontamination parameters (ie. time and temperature) must be defined and must be effective against the microorganisms of concern. The internal temperature and pressure of the effluent tanks and decontamination time must be logged throughout the cycle. Chemical-based decontamination systems may be practical on a small-scale where smaller volumes of liquid effluent require treatment.

Decontaminated liquids released from the treatment system must meet all applicable regulations (e.g. municipal bylaws for temperature, chemical/metal content, suspended solids, oil/grease and biochemical oxygen demand).

Carcass disposal systems:

Animal anatomical wastes may be incinerated. Effective incineration depends on proper equipment design; provision for the time, temperature, turbulence, and air required for complete oxidation; and careful feeding of the unit. Other technologies proven to be effective in decontamination (e.g. tissue autoclaves) present an acceptable alternative to incineration. The tissue autoclave parameters (ie. time, temperature and pressure) must be defined and logged throughout the cycle. Provincial and territorial regulatory requirements for incinerator operation/emissions and tissue autoclave discharges must be followed.

Radiation:

Gamma irradiation (e.g. ⁶⁰cobalt) can be used for the decontamination of heat-sensitive materials and is an effective means of decontaminating chemicals and solvents removed from a containment facility. The efficacy of the treatment technology is dependent on the penetration of the treated items by gamma irradiation and therefore, it is dependent upon the density of the treated substance as well as the strength of the irradiation source.

Microwave radiation is not widely used for decontamination in containment facilities. As in steam autoclaving, heat is the critical factor for eliminating viable microorganisms and the autoclave is usually the technology of choice. The factors which affect microwave treatment include frequency and wavelength of the irradiation, the duration of exposure and the destruction and moisture content of the material to be decontaminated.

Ultraviolet radiation (UV) should not be relied upon as the sole method of decontamination for materials removed from containment facilities (UV does not penetrate, microorganisms vary in susceptibility to UV). It is effective in reducing airborne and surface contamination providing the lamps are properly cleaned, maintained and checked to ensure that the appropriate intensity is being emitted.

7. Certification

For the purposes of this document, "certification" is defined as the verification of the physical construction and performance of critical containment components. Certification of containment systems may be included as part of the overall commissioning processes normally undertaken to verify that the design meets applicable codes and standards and that it has been constructed in accordance with the design intent.

To ensure the physical requirements for the intended containment level and use of the facility have been met, each laboratory and animal facility must undergo a detailed certification regiment. This requires verification and documentation of critical containment components. A complete set of "as built" and "as modified" drawings, 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 required before certification can proceed.

A checklist of critical containment components to be verified during initial certification is provided below. Re-certification should also be carried out on a regular basis to monitor the system's performance. The frequency of recertification depends on a variety of factors including the frequency of use and can only be determined by monitoring the facility over time. Initially, re-certification should be carried out on an annual basis. A comparison should be made to the baseline established during initial certification. Detailed records of the certification process and test results must be maintained.

Operational protocols must also be established before work with pathogens at the specified containment level can be carried out. Training of personnel is a critical aspect of this process and may involve initial work with pathogens normally requiring a lower containment level. Users must understand the containment systems and their operation in addition to scientific procedures.

7.1 Room Integrity

Smoke testing the integrity of a containment room can be used to detect any visual leaks in the room perimeter. All joints, corners and sealed penetrations should be surveyed for leaks. Pressure decay testing the integrity of the containment room provides an indication of the tightness of the room perimeter (ie. the ability of gases and liquids to move through the perimeter membrane and service penetrations).

The basic procedure for room pressure decay testing under vacuum is as follows:

isolate area by closing and securing all doors, valves and bubble tight dampers at the containment barrier (avoid temporary sealing measures in doors, windows and services that would cover permanent seals and not permit their testing for leakage); plug all pressure sensor lines, e.g. magnehelic gauges
connect a vacuum source to the room and create a negative pressure above 600 Pa negative pressure; allow room to stabilize and shut off vacuum
dynamically trend pressure loss starting at 600 Pa until the room reaches 200 Pa negative pressure; record at 10 second intervals for minimum of 30 minutes
record room temperature and exterior barometric pressure at beginning and end of test
acceptance criteria outlined below must be satisfied (note: sources of air leakage can be identified using a soap or commercial detector solution on joints, corners, sealed penetrations, etc.)

Testing requirements and acceptance criteria for certification of laboratories:

integrity of AP containment level 3 laboratories to be tested by smoke testing
integrity of AP containment level 4 laboratories to be tested by pressure decay testing; rate of air leakage not to exceed 12.5 Pa/min at 500 Pa over a twenty minute period

Testing requirements and acceptance criteria for certification of small animal facilities:

integrity of AP containment level 3 small animal facilities to be tested by smoke testing
pressure decay testing of AP containment level 3 animal rooms is recommended
pressure decay testing of AP containment level 3 animal rooms is required where animals are not housed in containment cages; rate of air leakage not to exceed 12.5 Pa/min at 500 Pa over a twenty minute period
integrity of AP containment level 4 animal rooms to be tested by pressure decay testing; rate of air leakage not to exceed 12.5 Pa/min at 500 Pa over a twenty minute period

Testing requirements and acceptance criteria for certification of large animal facilities:

room integrity of AP containment level 3 animal cubicles and PM room to be tested by smoke testing
pressure decay testing of AP containment level 3 animal cubicles and PM room is recommended
pressure decay testing of AP containment level 3 animal cubicles and PM room is required where non-indigenous agents are present; rate of air leakage not to exceed 12.5 Pa/min at 500 Pa over a twenty minute period
integrity of AP containment level 4 animal cubicles and PM room to be tested by pressure decay testing; rate of air leakage not to exceed 12.5 Pa/min at 500 Pa over a twenty minute period

7.2 Air Handling System

Various components of a containment room's air handling system require certification. Manufacturer's requirements for airflows for BSCs must be met. Particle challenge testing of HEPA filters must be performed to ensure they do not contain leaks in the filter media, the bond between the media and frame, or around the frame gasket and support. Ductwork systems should be pressure decay tested to confirm that specified leakage rates are not exceeded. The American Society of Mechanical Engineers (ASME) Standard N510 Testing of Nuclear Air Treatment Systems, 1989, gives procedures for testing the leak-tightness of ducts and plenums. Room pressure control systems must operate as specified (e.g. ensure negative pressures are maintained).

Biological safety cabinets:

all biological safety cabinets to be tested in-situ in accordance with CSA Z316.3-95, Biological Containment Cabinets: Installation and Field Testing (1995)
interlocks (ie. between BSC motor and exhaust fan) to be tested for specified response
alarms to be tested for detection of BSC and/or exhaust fan failure by simulation of alarm conditions

Fume hoods:

all fume hoods and associated exhaust systems to be tested in-situ in accordance with CSA Z316.5-94, Fume Hoods and Associated Exhaust Systems (1994)

HEPA filters and filter housings:

integrity of HEPA filters installed into supply and exhaust ductwork to be tested in-situ by particle challenge testing using the scanning method; particle penetration not to exceed 0.01%
integrity of HEPA filter housings with inlet and outlet bubble tight dampers installed into supply and exhaust ductwork to be tested in-situ by pressure decay testing in accordance with ASME N510 Testing of Nuclear Air Treatment Systems (1989); rate of air leakage not to exceed 0.2% of vol/min at 2500 Pa
in-line filters (e.g. plumbing vent lines, gas supply lines, autoclave exhaust ducts) to be tested by particle challenge testing; filter efficiency to be equivalent to HEPA

Supply and exhaust ductwork:

The basic procedure for ductwork pressure decay testing as outlined by ASME N510, Testing of Nuclear Air Treatment Systems, is as follows:

isolate and seal the ductwork to be tested
attach a temperature and pressure sensor to the test system to indicate interior temperature and pressure
pressurize ductwork with air to at least 500 Pa and allow to stabilize
record temperature inside the ductwork at beginning and end of the test
dynamically trend pressure loss; record at 10 second intervals for minimum of 30 minutes
the acceptance criteria outlined below must be satisfied (note: sources of air leakage can be identified using a soap or commercial detector solution on joints, corners, sealed penetrations, etc.)

Ductwork testing requirements and acceptance criteria:

for AP containment level 3 and 4 laboratories, SA rooms, and LA cubicles and PM rooms, supply and exhaust ductwork between containment room perimeter and bubble tight damper to be in accordance with Sheet Metal and Air Conditioning Contractors National Association (SMACNA) Seal Class A, HVAC Air Duct Leakage Test Manual (1985)
for AP containment level 3 SA rooms where animals are not housed in containment caging, LA cubicles and PM rooms where non-indigenous agents are present, supply and exhaust ductwork between containment room perimeter and bubble tight damper to be tested in-situ by pressure decay testing; rate of air leakage not to exceed 0.2% of duct vol/min at 500 Pa
for AP containment level 4 laboratories, SA rooms, and LA cubicles and PM rooms, supply and exhaust ductwork between containment room perimeter and bubble tight damper to be tested in-situ by pressure decay testing; rate of air leakage not to exceed 0.1% duct vol/min at 500 Pa

Air balancing and pressurization relationships:

inward directional airflow to be visually demonstrated by smoke testing
pressurization relationships across a containment barrier (e.g. at airlocks, laboratories, animal rooms and cubicles, corridors) to be verified; minimum of 25 Pa is recommended across a containment barrier

HVAC control systems:

control systems to be tested for fail-safe operation by failure of system components (e.g. door is open, fan failure, electrical failure, BSC failure); room/cubicle positive pressurization to be prevented
control system performance verification should include speed of response, accuracy, and repeatability
audible alarms to be tested for detection of positive pressurization and air handling systems failure by simulation of alarm conditions

7.3 Laboratory Services

water supply backflow prevention to be tested in accordance with CAN/CSA-B64.10-94, Manual for the Selection, Installation, Maintenance, and Field Testing of Backflow Prevention Devices (1994)
backflow prevention for other services (e.g. gases) to be verified to ensure system will operate as specified
the water flow rate and temperature of eyewash stations and emergency showers to be verified in accordance with ANSI Z3858.1, Emergency Eyewash and Shower Equipment
compressed breathing air and systems to be verified in accordance with CAN3-Z180.1-M85, Compressed Breathing Air and Systems (1994)
operation of positive-personal protective equipment (ie. suit) to be tested in-situ to ensure suit will operate as specified
water and chemical shower systems to be tested to ensure systems will operate as specified
cage washers to be tested to ensure system will operate as specified; water temperature of final rinse to be at least 82ºC
emergency electrical generator to be tested under appropriate load conditions to ensure systems will operate as specified
security systems (e.g. controlled access, closed circuit TV) to be verified to ensure system will operate as specified
communication and electronic paper transfer systems (e.g. intercom, telephone, fax) to be verified to ensure system will operate as specified
all sterilization systems (e.g. autoclaves, liquid effluent treatment systems) to be verified for operation as specified and to be microbiologically tested using representative loads; for technologies based on heat - using Bacillus stearothermophilus spores; for technologies based on chemicals - using Bacillus subtilis spores
drains and associated piping leading to liquid effluent treatment systems (including associated vent lines) to be tested in accordance with Section 3.6 of the Canadian Plumbing Code, Testing of Drainage and Venting Systems (1990); pressure for air test on drainage system shall be at a factor of safety beyond standard code requirements of 35 kPa (e.g. 2 X code)
all disinfection systems (e.g. dunk tanks, fumigation chambers) to be verified for operation as specified and microbiologically tested using representative loads; resistance of test organism to be representative of organisms likely to be encountered

8. References

Biological Containment Cabinets: Installation and Field Testing. CSA Z316.3-95, Canadian Standards Association, Toronto, ON. 1995.

Canadian Plumbing Code. Associate Committee on the National Building Code. National Research Council of Canada, Ottawa, ON. 1990.

Class II Biohazard Cabinetry. Standard No. 49. National Sanitation Foundation, Ann Arbor, MI. 1992.

Compressed Breathing Air and Systems. CAN3.Z180.1-M85. Canadian Standards Association, Toronto, ON. 1994.

Emergency Eyewash and Shower Equipment. ANSI Z358.1. American National Standards Institute, New York, NY. 1990.

Fume Hoods and Associated Exhaust Systems. Z316.5-94. Canadian Standards Association, Toronto, ON. 1994.

Gravimetric and Dust-spot Procedures for Testing Air-Cleaning Devices Used in General Ventilation for Removing Particulate Matter. Standard 52.1. American Society of Heating, Refrigerating, and Air-conditioning Engineers, Inc., Atlanta, GA. 1992.

Guide for the Care and Use of Experimental Animals. Canadian Council on Animal Care, Ottawa, ON. 1993.

Heating, Ventilating, and Air-Conditioning Applications. American Society of Heating, Refrigeration and Air-Conditioning Engineers, Inc., Atlanta, Ga. 1995.

HVAC Air Duct Leakage Test Manual. Sheet Metal and Air Conditioning Contractors National Association, Inc., Chantilly, Virginia. 1985.

Laboratory Biosafety Guidelines. Health Canada. Supply and Services Canada, Ottawa, ON. 1996.

Manual for the Selection, Installation, Maintenance, and Field Testing of Backflow Prevention Devices. CAN/CSA-B64.10-94. Canadian Standards Association, Toronto, ON. 1994

Method of Testing Performance of Laboratory Fume Hoods. American Society of Heating, Refrigerating and Air-conditioning Engineers, Inc., Atlanta, GA. 1995.

National Building Code of Canada. Canadian Commission on Building and Fire Codes/National Research Council of Canada. Ottawa, ON. 1995

Primary Containment for Biohazards: Selection, Installation, and Use of Biological Safety Cabinets. Centers for Disease Control/National Institutes of Health. U.S. Government Printing Office, Washington, D.C. 1995

Recommended Practice for HEPA Filters. IES-RP-CC-OO1-86. Institute of Environmental Sciences, Mount Prospect, IL. 1986.

Testing of Nuclear Air Treatment Systems. ASME N510. American Society of Mechanical Engineers, New York, NY. 1989.

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