Canadian Biosafety Standard-Eng

Canadian Biosafety Standard, Second Edition
is available on the Internet at the following address:

http://canadianbiosafetystandards.collaboration.gc.ca/
galement disponible en franais sous le titre :
Norme canadienne sur la bioscurit, deuxime dition
To obtain additional copies, please contact:
Public Health Agency of Canada
100 Colonnade Road
Ottawa, ON K1A 0K9
Tel.: 613-957-1779
Fax.: 613-941-0596
PHAC email: [email protected]
CFIA email: [email protected]
This publication can be made available in alternative formats upon request.
Her Majesty the Queen in Right of Canada, as represented by the Minister of Health
and the Minister of Agriculture and Agri-Food, 2015
Publication date: March 2015
This publication may be reproduced for personal or internal use only without permission
provided the source is fully acknowledged.
Print Cat.: HP45-7/2015E
ISBN: 978-1-100-25771-6
Publication Number: 140467
PDF Cat.: HP45-7/2015E-PDF

ISBN: 978-1-100-25772-3
TABLE OF CONTENTS
PREFACE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . iii
ABBREVIATIONS AND ACRONYMS. . . . . . . . . . . . . . . . . vii
GLOSSARY . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xi
NORMATIVE REFERENCES . . . . . . . . . . . . . . . . . . . . . xxxi
CHAPTER 1 INTRODUCTION. . . . . . . . . . . . . . . . . . . . . 1
1.1 Scope. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
1.2 Regulatory Authorities. . . . . . . . . . . . . . . . . . . . . . . . . . . 3
1.3 Legislative and Regulatory Requirements for
HumanPathogens and Toxins. . . . . . . . . . . . . . . . . . . . . . 4
1.4 Legislative and Regulatory Requirements for
AnimalPathogens, Toxins, and other Regulated
InfectiousMaterial. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
CHAPTER 2 HOW TO USE THE

CANADIANBIOSAFETYSTANDARD. . . . . . 13
2.1
2.2
2.3
2.4
2.5
Abbreviations, Definitions, and References. . . . . . . . . . . 14

Containment Levels and Containment Zones. . . . . . . . . . 14
Working with Human and Animal Pathogens and Toxins. . 15
Working with Animals . . . . . . . . . . . . . . . . . . . . . . . . . . 19
Derogations and Exemptions to Physical
or Operational Requirements. . . . . . . . . . . . . . . . . . . . . 20
2.6 Matrix Layout: Chapters 3, 4, and 5 . . . . . . . . . . . . . . . 20
2.7 Explanatory Notes . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24
2.8 The Canadian Biosafety Handbook . . . . . . . . . . . . . . . . 24
CHAPTER 3 PHYSICAL CONTAINMENT

REQUIREMENTS. . . . . . . . . . . . . . . . . . . 25
3.1
Structure and Location. . . . . . . . . . . . . . . . . . . . . . . . . .
3.2 Containment Barrier . . . . . . . . . . . . . . . . . . . . . . . . . . .
3.3 Access . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
3.4 Surface Finishes and Casework . . . . . . . . . . . . . . . . . . .
3.5 Air Handling. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
3.6 Facility Services. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
3.7 Essential Biosafety Equipment. . . . . . . . . . . . . . . . . . . . .
3.8 Effluent Decontamination Systems. . . . . . . . . . . . . . . . . .
26
27
29
34
36
39
42
46
CHAPTER 4 OPERATIONAL PRACTICE

REQUIREMENTS. . . . . . . . . . . . . . . . . . . 49
4.1
4.2
4.3
4.4
4.5
4.6
4.7
4.8
4.9
4.10
Biosafety Program Management . . . . . . . . . . . . . . . . . .

Medical Surveillance Program. . . . . . . . . . . . . . . . . . . .
Training Program. . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Personal Protective Equipment . . . . . . . . . . . . . . . . . . . .
Entry and Exit of Personnel, Animals, and Material. . . . .
Work Practices . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Animal Work Considerations. . . . . . . . . . . . . . . . . . . . .
Decontamination and Waste Management. . . . . . . . . . .
Emergency Response. . . . . . . . . . . . . . . . . . . . . . . . . . .
Records and Documentation. . . . . . . . . . . . . . . . . . . . . .
50
57
58
60
62
66
73
75
78
81
CHAPTER 5 PERFORMANCE AND VERIFICATION

TESTING REQUIREMENTS. . . . . . . . . . . . 85
5.1
5.2
5.3
Performance and Verification Tests for All

Containment Levels. . . . . . . . . . . . . . . . . . . . . . . . . . . . 86
Additional Performance and Verification Tests for
Select CL2 and CL2- Ag Zones and All CL3-CL4 Zones . . . 89
Performance and Verification Tests to be Conducted
onCommissioning of Containment Zone. . . . . . . . . . . . . 93
REFERENCES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95
APPENDIX . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 99
PREFACE
PREFACE
The Government of Canadas Canadian Biosafety Standard (CBS), 2nd Edition, 2015, is
a harmonized national standard for the handling or storing of human and terrestrial animal
pathogens and toxins in Canada. Activities in Canada involving human and animal
pathogens and toxins are regulated by the Public Health Agency of Canada (PHAC) and
the Canadian Food Inspection Agency (CFIA) in accordance with the Human Pathogens
and Toxins Act (HPTA), the Human Pathogens and Toxins Regulations (HPTR), the Health of
Animals Act, and the Health of Animals Regulations.
The second edition of the CBS updates the biosafety standard originally published as Part I
of the Canadian Biosafety Standards and Guidelines (CBSG), 1st Edition, 2013. The CBSG
was developed to update and harmonize three existing Canadian biosafety standards and
guidelines for the design, construction, and operation of facilities in which pathogens or
toxins are handled or stored:
1.
Human pathogens and toxins: Laboratory Biosafety Guidelines, 3rd Edition,

2004 (PHAC)
2.
Terrestrial animal pathogens: Containment Standards for Veterinary Facilities,

1st Edition, 1996 (CFIA)
3.
Prions: Containment Standards for Laboratories, Animal Facilities and Post Mortem
Rooms Handling Prion Disease Agents, 1st Edition, 2005 (CFIA)
The CBS sets out the physical containment, operational practice, and performance and
verication testing requirements for the safe handling or storing of human and terrestrial animal
pathogens and toxins. The CBS updates many requirements to be more risk-, evidence-, and
performance-based, as well as incorporating new information in the eld of biocontainment
engineering. In addition, the CBS includes several new requirements and information to
support the full implementation of the HPTA and the HPTR. On December 1st, 2015, the HPTR
comes into force and the CBS will come into effect and supersede the CBSG. The CBS will be
used by the PHAC and the CFIA to verify the ongoing compliance of regulated facilities with
the applicable legislation. This will support licence applications, renewals, animal pathogen
import permit applications, and, where applicable, the facility certication (and recertication)
of containment zones.
iv
Canadian Biosafety Standard (CBS) 2nd Edition
The Guidelines originally published as Part II of the CBSG have also been updated and are
published in the Canadian Biosafety Handbook (CBH), 2nd Edition, 2015. The CBH is a
companion document to the CBS that provides core information and guidance as to how
the biosafety and biosecurity requirements outlined in the CBS can be achieved. The CBH
systematically addresses the concepts required for the development and maintenance of a
comprehensive risk-based biosafety management program. In some instances, however, best
practices or risk mitigation strategies other than those described in the CBH may also be
acceptable to achieve the requirements of the CBS.
The PHAC and the CFIA welcome comments, clarications, and suggestions for incorporation
into future editions of the CBS. To this end, please send information, with references (where
applicable), for the continual improvement of the CBS to:
PHAC email: 1)"$standardsnormes"[email protected]
CFIA email: [email protected]
ABBREVIATIONS AND ACRONYMS
ABBREVIATIONS AND ACRONYMS

Ag
Agriculture (i.e., CL2-Ag, CL3-Ag)
AIRS
Automated Import Reference System
ANSI
American National Standards Institute
ASME
American Society of Mechanical Engineers
BSC
Biological safety cabinet
BSO
Biological safety ofcer
CAN
National Standard of Canada
CBH
Canadian Biosafety Handbook
CBS
Canadian Biosafety Standard
CBSG
Canadian Biosafety Standards and Guidelines
CFIA
Canadian Food Inspection Agency
CL
Containment level (i.e., CL1, CL2, CL3, CL4)
CSA
Canadian Standards Association
CSFHAAP
Containment Standards for Facilities Handling Aquatic Animal Pathogens
CSFHPP
Containment Standards for Facilities Handling Plant Pests
ERP
Emergency response plan
HAA
Health of Animals Act
HAR
Health of Animals Regulations
HEPA
High efciency particulate air
HPTA
Human Pathogens and Toxins Act
viii
HPTA
4ecurity
$learance
Human Pathogens and Toxins Act 6ecurity &learance
HPTR
Human Pathogens and Toxins Regulations
HVAC
Heating, ventilation, and air conditioning
IDA
Inward directional airow
IEST
Institute of Environmental Sciences and Technology
in. w.g.
Inches of water gauge (unit of pressure; 1 in. w.g. = 250 Pa)
LA zone
Large animal containment zone
LRA
Local risk assessment
NSF
National Sanitation Foundation
PHAC
Public Health Agency of Canada
PM room
Post mortem room
PPE
Personal protective equipment
PSDS
Pathogen safety data sheet
RG
Risk group (i.e., RG1, RG2, RG3, RG4)
SA zone
Small animal containment zone
SMACNA
Sheet Metal and Air Conditioning Contractors National Association
SOP
Standard operating procedure
SSBA
Security sensitive biological agent
UPS
UninterruptJCMe power supply
ix
GLOSSARY
GLOSSARY
It is important to note that while some of the denitions provided in the glossary are
universally accepted, many of them were developed specically for the Canadian Biosafety
Standard (CBS); therefore, some denitions may not be applicable to facilities that fall
outside of the scope of the CBS.
Accident
An unplanned event that results in injury, harm, or damage.
Administrative
area
Dedicated room or adjoining rooms that are used for activities

that do not involve infectious material and toxins. Administrative
areas do not require any containment equipment, systems, or
operational practices. Examples of administrative areas include
ofces, photocopy areas, and meeting/conference rooms.
Aerosol
A suspension of ne solid particles or liquid droplets in a gaseous

medium (e.g., air) that can be created by any activity that imparts
energy into a liquid/semi-liquid material.
Airborne
pathogen
A pathogen that is capable of moving through or being carried

by the air.
Airtight doors
Doors that are designed to allow no leakage of air (0%) under

normal operating conditions and to withstand pressure decay
testing and gaseous decontamination. Airtight doors can be
achieved with inatable or compression seals.
Animal cubicle
A room or space designed to house an animal (or animals) where

the room itself serves as primary containment. These spaces are
used to house large-sized animals (e.g., livestock, deer), or smallsized animals that are housed in open caging (i.e., not primary
containment caging).
Animal pathogen
Any pathogen that causes disease in animals; including those

derived from biotechnology. In the context of the Canadian
Biosafety Standard, animal pathogen refers only to pathogens
that cause disease in terrestrial animals; including those that infect
avian and amphibian animals, but excluding those that cause
disease in aquatic animals and invertebrates.
xii
Animal pathogen
import permit
A permit issued by the Public Health Agency of Canada or

the Canadian Food Inspection Agency for the importation
into Canada of: animal pathogens or toxins; animals, animal
products, animal by-products, or other organisms carrying an
animal pathogen or part of one; under Section 51(a) and (b) of
the Health of Animals Regulations.
Animal room
A room designed to house animals in primary containment

caging. These spaces are used to house only small-sized animals
(e.g., mice, rats, rabbits).
Anteroom
A room, or series of rooms, inside the containment zone, used

to separate clean areas from dirty areas (i.e., area with a
lower risk of contamination from those with a higher risk of
contamination), for personnel and animal entry/exit across the
containment barrier, and for entry to/exit from animal rooms,
animal cubicles, and post mortem rooms. The negative differential
air pressures required in containment zones where inward
directional airow is provided can be more effectively maintained
through the presence of an anteroom. An anteroom may also
provide appropriate space at the entry/exit point(s) to don,
doff and store dedicated containment zone clothing and
additional personal protective equipment, as required.
Authorized
personnel
Individuals who have been granted unsupervised access to the

containment zone by the containment zone director, biological safety
ofcer, or another individual to whom this responsibility has been
assigned. This is dependent on completing training requirements and
demonstrating prociency in the standard operating procedures, as
determined to be necessary by the facility.
Backdraft
protection
A system that protects the air supply to the containment zone

from contamination in the event of a reversal of airflow. High
efciency particulate air (HEPA) lters or isolation dampers are
commonly used to prevent contamination from reaching areas of
lower containment.
;Z\dhp
prevention
A system that protects the water supply to the containment zone

from contamination. Many types of backow devices also have
test ports so that they can be checked to ensure that they are
functioning properly.
xiii
GLOSSARY
Biocontainment
See containment.
Biological
material
Pathogenic and non-pathogenic microorganisms, proteins,

and nucleic acids, as well as any biological matter that may
contain microorganisms, proteins, nucleic acids, or parts thereof.
Examples include, but are not limited to, bacteria, viruses, fungi,
prions, toxins, genetically modied organisms, nucleic acids,
tissue samples, diagnostic specimens, live vaccines, and isolates
of a pathogen (e.g., pure culture, suspension, puried spores).
Biological safety
cabinet (BSC)
A primary containment device that provides protection for personnel,

the environment, and the product (depending on BSC class), when
working with biological material.
Biological safety
h_\^k!;LH"
An individual designated for overseeing the facilitys biosafety and

biosecurity practices.
Biosafety
Containment principles, technologies, and practices that are

implemented to prevent unintentional exposure to infectious
material and toxins, or their accidental release.
Biosafety Manual
A facility-specic manual that describes the core elements of a

biosafety program (e.g., biosecurity plan, training, personal
protective equipment).
Biosecurity
Security measures designed to prevent the loss, theft, misuse,

diversion, or intentional release of pathogens, toxins, and other
related assets (e.g., personnel, equipment, non-infectious material,
and animals).
Biosecurity risk
assessment
A risk assessment in which the pathogens, toxins, infectious material,

and other related assets (e.g., equipment, animals, information)
in possession are identied and prioritized, the threats and risks
associated with these materials are dened, and appropriate
mitigation strategies are determined to protect these materials
against potential theft, misuse, diversion, or intentional release.
xiv
Clean
change area
The designated space where dedicated personal protective

equipment is donned when entering the containment zone,
animal cubicle, or post mortem room. The clean change area
is considered to be free from contamination when entry and exit
procedures are routinely followed. In high containment zones, the
clean change area is located outside the containment barrier.
Closed system
An apparatus or process system designed to contain biological

material and prevent its release into the surrounding environment.
Commissioning
A process whereby a newly constructed containment zone, or a

newly modied or renovated containment zone, is subjected to
a series of performance and verication tests to ensure that the
nished containment zone, including equipment and containment
systems, will operate in accordance with the physical design
intent and specications and is ready to be put into operation, or
resume activities involving pathogens and toxins, respectively.
Community
Encompasses both human (i.e., the public) and animal populations.
Containment
The combination of physical design parameters and operational

practices that protect personnel, the immediate work environment,
and the community from exposure to biological material. The term
biocontainment is also used in this context.
Containment
barrier
The boundary between clean and dirty areas (i.e., between

the laboratory work areas, animal rooms, animal cubicles, or post
mortem rooms, and outside of that containment area). Where
inward directional airow is provided, a physical containment
barrier of air is established to protect against airborne or aerosolized
infectious material or toxins from reaching the clean areas.
Containment
level (CL)
Minimum physical containment and operational practice

requirements for handling infectious material or toxins safely in
laboratory, large scale production, and animal work environments.
There are four containment levels ranging from a basic laboratory
(containment level 1 [CL1]) to the highest level of containment
(containment level 4 [CL4]).
xv
GLOSSARY
Containment
system
Dedicated equipment that functions to provide and maintain

containment. This includes, but is not limited to, primary containment
devices (e.g., biological safety cabinets), heating, ventilation, and
air conditioning (HVAC) and control systems, and decontamination
systems (e.g., autoclaves).
Containment zone
A physical area that meets the requirements for a specied

containment level. A containment zone can be a single room
(e.g., containment level 2 [CL2] laboratory), a series of co-located
rooms (e.g., several non-adjoining but lockable CL2 laboratory
work areas), or it can be comprised of several adjoining rooms
(e.g., containment level 3 [CL3] suite with dedicated laboratory
areas and separate animal rooms, or animal cubicles). Dedicated
support areas, including anterooms (with showers and clean
and dirty change areas, where required), are considered to be
part of the containment zone.
Contamination
The undesired presence of infectious material or toxins on a

surface (e.g., benchtop, hands, gloves) or within other materials
(e.g., laboratory samples, cell cultures).
Controlled access
system
A physical or electronic system designed to restrict access to

authorized personnel only.
Controlled
activities
Any of the following activities referred to in Section 7(1) of the

Human Pathogens and Toxins Act: possessing, handling or using
a human pathogen or toxin; producing a human pathogen or
toxin; storing a human pathogen or toxin; permitting any person
access to a human pathogen or toxin; transferring a human
pathogen or toxin; importing or exporting a human pathogen or
toxin; releasing or otherwise abandoning a human pathogen or
toxin; or disposing of a human pathogen or toxin.
Critical door
Any door directly located on the containment barrier of a

containment zone, animal cubicle, or post mortem room where
inward directional airow is required.
xvi
Culture
The in vitro propagation of microorganisms, tissue cells, or other

living matter under controlled conditions (e.g., temperature,
humidity, nutrients) to generate greater numbers or a higher
concentration of the organisms/ cells. In the context of the
Canadian Biosafety Standard, cell culture refers to cells derived
from a human or animal source.
Decontamination
The process by which materials and surfaces are rendered

safe to handle and reasonably free of microorganisms, toxins,
or prions; this may be accomplished through disinfection,
inactivation, or sterilization.
Decontamination
technology
Equipment proven by validation to render materials safe to

handle and reasonably free of microorganisms, toxins, or prions.
Examples include autoclaves, incinerators, tissue digesters, and
efuent decontamination systems.
Deep seal trap
A plumbing drain trap that has an effective head or depth that is

sufcient to maintain a water seal, in accordance with air pressure
differentials (i.e., water is neither siphoned into the room nor
pushed through the trap). These traps have a water seal greater
than 102 mm (4 inches) in depth, and a trap seal of 127 mm to
152 mm (5 to 6 inches).
Dirty
change area
The designated space inside the containment barrier where

contaminated personal protective equipment is doffed when
exiting the containment zone, animal cubicle, or post mortem
room. The dirty change area is considered to be contaminated
or potentially contaminated during normal operations.
Disease
A disorder of structure or function in a living human or animal,

or one of its parts, resulting from infection or intoxication. It is
typically manifested by distinguishing signs and symptoms.
Disinfection
Process that eliminates most forms of living microorganisms;

disinfection is much less lethal to infectious material than sterilization.
Dual-use potential
Qualities of a pathogen or toxin that allow it to be either used

for legitimate scientic applications (e.g., commercial, medical,
or research purposes), or intentionally misused as a biological
weapon to cause disease (e.g., bioterrorism).
xvii
GLOSSARY
Dunk tank
A disinfectant-lled vessel located at or on the containment

barrier that allows for the safe removal of material and samples
from containment zones via surface decontamination achieved
through immersion.
>_n^gm
decontamination
system
Equipment connected to the drain plumbing used to

decontaminate, through heat or chemical means, the liquid waste
(i.e., efuent) produced in a containment zone prior to release
into sanitary sewers.
Emergency
Response Plan
(ERP)
A document outlining the actions to be taken and the parties

responsible in emergency situations such as a spill, exposure,
release of infectious material or toxins, animal escape, personnel
injury or illness, power failure, re, explosion, or other emergency
situations (e.g., ood, earthquake, hurricane).
Emerging
animal disease
A new infectious disease resulting from the evolution or change

of an existing pathogenic agent, a known infectious disease
spreading to a new geographic area or population, or a
previously unrecognized pathogenic agent or disease diagnosed
for the rst time and which has a signicant impact on animal
health. Emerging animal disease pathogens are handled as
non-indigenous animal pathogens due to the high risk of serious
negative effects associated with these pathogens.
Exporting
The activity of shipping (e.g., transferring or transporting)

pathogens, toxins, or other regulated infectious material from
Canada to another country.
Exposure
Contact with, or close proximity to, infectious material or toxins

that may result in infection or intoxication, respectively. Routes of
exposure include inhalation, ingestion, inoculation, and absorption.
Exposure
follow-up report
A tool used to report and document incident occurrence and

investigation information for an exposure incident previously
notied to the Public Health Agency of Canada.
Exposure
ghmb\Zmbhg
report
A tool used to notify and document preliminary information to the

Public Health Agency of Canada of an exposure incident.
xviii
Facility
(plural: facilities)
Structures or buildings, or dened areas within structures or

buildings, where infectious material or toxins are handled or
stored. This could include individual research and diagnostic
laboratories, large scale production areas, or animal housing
zones. A facility could also be a suite or building containing more
than one of these areas.
Facility
\^kmb\Zmbhg
The formal acknowledgement from the Canadian Food Inspection

Agency (CFIA) that a containment zone or facility where imported
animal pathogens will be handled or stored complies with the
physical containment, operational practice, and performance
and verication testing requirements described in the Canadian
Biosafety Standard. Recertication refers to the renewal of the
facility certication issued by the CFIA following a streamlined
review process.
Good
microbiological
laboratory
practices
A basic laboratory code of practice applicable to all types of

activities with biological material. These practices serve to protect
workers and prevent contamination of the environment and the
samples in use.
Gross
contamination
The accumulation of organic material (e.g., bedding, feed,

excrement, blood, and tissues) on a surface that can be removed
by physical methods, such as scraping, brushing, and wiping.
Handling
or storing
Handling or storing pathogens, toxins, or infectious material

includes possessing, handling, using, producing, storing,
permitting access to, transferring, importing, exporting, releasing,
disposing of, or abandoning such material. This includes all
controlled activities involving human pathogens and toxins
specied in Section 7(1) of the Human Pathogens and Toxins Act.
High
concentration
Infectious material or toxins that are concentrated to a degree that

increases the risks associated with manipulating the material (i.e.,
increases the likelihood or consequences of exposure).
High containment
zones
Containment zones (i.e., laboratory work areas, animal

rooms and cubicles, post mortem rooms, areas for large scale
production), including all dedicated support areas, in containment
level 3 (CL3), containment level 3-Agriculture (CL3-Ag), and
containment level 4 (CL4).
xix
GLOSSARY
Abà^_\b^g\r
particulate air
!A>I:"em^k
A device capable of ltering 99.97% of airborne particles 0.3

m in diameter, the most penetrating particle size. Due to the
effects of impaction, diffusion, and interception, HEPA lters are
even more efcient at trapping and retaining particles that are
either smaller or larger than 0.3 m in diameter.
Human Pathogens
and Toxins Act
4ecurity $learance
(HPTA 4ecurity
$learance)
An authorization following verication of an individuals

background and reliability status issued by the Public Health
Agency of Canada under Section 34 of the Human Pathogens
and Toxins Act.
Importing
The activity of bringing (e.g., transferring or transporting)

pathogens, toxins, or other regulated infectious material into
Canada from another country.
Incident
An event or occurrence with the potential of causing injury, harm,

infection, intoxication, disease, or damage. Incidents can involve
infectious material, infected animals, or toxins, including a spill,
exposure, release of infectious material or toxins, animal escape,
personnel injury or illness, missing infectious material or toxins,
unauthorized entry into the containment zone, power failure,
re, explosion, ood, or other crisis situations (e.g., earthquake,
hurricane). Incidents include accidents and near misses.
Infectious material
Any isolate of a pathogen or any biological material that contains

human or animal pathogens and, therefore, poses a risk to human
or animal health.
In situ
Latin for on site or in place; describes a xed location at which

a procedure or experiment is conducted.
Interlock
A device or mechanism for coordinating the function of components

(e.g., to prevent two doors being open simultaneously, or to ensure
a supply fan shuts down in the event of an exhaust fan failure).
xx
Intoxication
A substance-induced disorder or disease resulting in a symptomatic

or asymptomatic condition, or other physiological change
resulting from an exposure (i.e., ingestion, inhalation, inoculation,
or absorption) to a toxin produced by or isolated from a
microorganism. This includes a similar response from exposure to a
synthetically produced microbial toxin.
Inventory
A list of (biological) assets associated with a containment zone

identifying pathogens, toxins, and other infectious material in
storage both inside and outside of the containment zone.
In vitro
Latin for within glass; describes experimentation involving

components of a living organism within an articial environment
(e.g., manipulation of cells in petri dish), including activities
involving cell lines or eggs.
In vivo
Latin for within the living; describes experimentation conducted

within the whole living organism (e.g., studying the effect of
antibiotic treatment in animal models).
Inward directional
Zbkhp!B=:"
Air that always ows from areas of lower containment or lower

contamination risk to areas of higher containment or higher
contamination risk, as the result of a negative air pressure differential
within the containment zone created by a ventilation system.
Isolation damper
A shut-off valve used to seal off air supply and exhaust air
ductwork to/from a containment zone, as well as plumbing vent
lines to allow the decontamination of high efciency particulate
air (HEPA) lters. Isolation dampers also provide backdraft
protection in the event of heating, ventilation, and air conditioning
(HVAC) system failure or a reversal of airow, and prevent puffback in certain types of biological safety cabinets.
Laboratory
An area within a facility or the facility itself where biological

material is handled for scientic or medical purposes.
Laboratory
work area
Area inside a containment zone designed and equipped for

in vitro research, diagnostics, and teaching purposes.
xxi
GLOSSARY
Large animal
containment zone
(LA zone)
Animal containment zone comprised of two or more co-located

or adjoining rooms of equal containment level where animals
are housed in animal cubicles (i.e., the room itself provides the
primary containment). An LA zone may include, for example,
large-sized animals, such as livestock or deer, housed in cubicles
or, cubicles where small-sized animals, such as mice or raccoons,
are housed in open caging (i.e., not primary containment
caging). Post mortem rooms, where present, are considered to be
part of an LA zone.
Large scale
Activities generally involving volumes of toxins or the in vitro

culture of infectious material on a scale of 10 litres or greater. This
could be a single vessel with a volume of 10 litres or greater, or
based on the processes and pathogen used, could be multiple
vessels with a total volume of 10 litres or greater. It is determined
in consultation with the Public Health Agency of Canada and/or
the Canadian Food Inspection Agency on a case-by-case basis,
whether or not particular activities conducted in a containment
zone are required to follow the increased or unique requirements
for large scale production areas.
Large-sized
animal
Refers to the physical size of the animal; large-sized animals are

generally too large to be housed in primary containment caging,
and are therefore housed in an animal cubicle. Examples include
cows, horses, moose, deer, and sheep.
Large volume
A volume of infectious material or toxins that is sufciently large to

increase the risk associated with the manipulation of the material (i.e.,
increases the likelihood or consequences of exposure or release).
Licence
An authorization to conduct one or more controlled activities with

human pathogens or toxins issued by the Public Health Agency of
Canada under Section 18 of the Human Pathogens and Toxins Act.
Limited access
Access that is only permitted to authorized personnel and other

authorized visitors through either operational means (e.g., having
authorized personnel actively monitor and check all individuals
entering a designated area) or through the use of a physical
barrier (e.g., a controlled access system, such as key-locks or
electronic access card).
xxii
Local risk
assessment (LRA)
Site-specic risk assessment used to identify hazards based

on the infectious material or toxins in use and the activities
being performed. This analysis provides risk mitigation and risk
management strategies to be incorporated into the physical
containment design and operational practices of the facility.
Long-term storage
In the context of the Canadian Biosafety Standard, the possession

of material (i.e., pathogens, toxins, and other regulated infectious
material) beyond 30 days of receipt or creation.
Medical
surveillance
program
A program designed to prevent and detect personnel illness

related to exposure to infectious material or toxins. The focus
of the program is primarily preventive, but provides a response
mechanism through which a potential infection or intoxication can
be identied and treated before serious injury or disease occurs.
Microorganism
A cellular or non-cellular microbiological entity, capable of

replication or transferring genetic material and that cannot be
reasonably detected by the naked human eye. Microorganisms
include bacteria, fungi, viruses, and parasites, and may be
pathogenic or non-pathogenic in nature.
Movement
The action of moving (e.g., bringing, carrying, leading,

relocating) people, material (including infectious material or
toxins), or animals from one physical location to another physical
location in the same building. This can include movement within
the same containment zone, to a different containment zone, or to
another location within the same building.
Non-indigenous
animal pathogen
A pathogen that causes an animal disease listed in the World

OrganiVation for Animal Healths OIE-Listed diseases, infections
and infestations (as amended from time to time) and that is exotic
to Canada (i.e., foreign animal disease agents that are not
present in Canada). These pathogens may have serious negative
health effects to the Canadian animal population.
Open caging
Caging intended to restrict animals to an area (e.g., animal pens).

This type of caging does not prevent the release of pathogens
and toxins and, therefore, does not meet the requirements for
primary containment caging.
xxiii
GLOSSARY
Operational
practice
requirements
Administrative controls and procedures followed in a containment

zone to protect personnel, the environment, and ultimately the
community, from infectious material or toxins, as outlined in
Chapter 4.
Overarching
risk assessment
A broad risk assessment that supports the biosafety program as a

whole and may encompass multiple containment zones within an
institution or organization. Mitigation and management strategies
reect the type of biosafety program needed to protect personnel
from exposure and to prevent the release of pathogens and toxins.
Pass-through
chamber
Interlocked double-door compartment situated on a containment

barrier that allows the safe movement of materials into and out of
containment zones.
Pass-through
technology
Equipment with double-door compartments situated on a

containment barrier that allows the safe movement of materials
into and out of the containment zone. Examples include doubledoor barrier autoclaves, pass-through chambers, dunk tanks,
barrier cage washers, and feed chutes.
Pathogen
A microorganism, nucleic acid, or protein capable of causing

disease or infection in humans or animals. Examples of human
pathogens are listed in Schedules 2 to 4 and in Part 2 of
Schedule 5 of the Human Pathogens and Toxins Act, but these are
not exhaustive lists. Examples of animal pathogens can be found
through the Automated Import Reference System on the Canadian
Food Inspection Agency website.
Pathogen risk
assessment
The determination of the risk group and appropriate physical

containment and operational practice requirements needed to
safely handle the infectious material or toxins in question.
Pathogenicity
The ability of a pathogen to cause disease in a human or animal host.
Performance and
o^kb\Zmbhgm^lmbg`
requirements
Performance and verication tests that are necessary to demonstrate

compliance with the physical containment requirements, as
outlined in Chapter 3 and, in some cases, the operational practice
requirements, as outlined in Chapter 4. The performance and
verication testing requirements are listed in Chapter 5.
xxiv
Personal
protective
equipment (PPE)
Equipment and/or clothing worn by personnel to provide a

barrier against infectious material or toxins, thereby minimizing the
risk of exposure. PPE may include, but is not limited to, lab coats,
gowns, full-body suits, gloves, protective footwear, safety glasses,
safety goggles, masks, and respirators.
Physical
containment
requirements
Physical barriers in the form of engineering controls and facility

design used to protect personnel, the environment, and ultimately
the community, from pathogens or toxins, as outlined in Chapter 3.
Post mortem room

(PM room)
A room within the containment zone where animal necropsies

and dissections are conducted.
Pressure decay
testing
A method of quantifying the leak rates of a sealed environment.
Primary
containment
The rst level of physical barriers designed to contain pathogens

and toxins and prevent their release. This is accomplished
by the provision of a device, equipment, or other physical
structure situated between the infectious material or toxins and
the individual, the work environment, or other areas within the
containment zone. Examples include biological safety cabinets,
glove boxes, and animal microisolators. In animal cubicles, the
room itself provides primary containment, and personal protective
equipment serves as primary protection against exposure.
Primary
containment
caging
Animal caging serving as a primary containment device to

prevent the release of infectious material and toxins. Examples
include ventilated lter-top cages and ventilated micro-isolator
cage rack system, with or without high efciency particulate air
(HEPA) lters.
Primary
containment
device
Apparatus or equipment that is designed to prevent the release of

infectious material or toxins and to provide primary containment
(i.e., provide a physical barrier between the individual and/or
the work environment and the biological material). Examples of
primary containment devices include biological safety cabinets,
isolators, centrifuges with sealable cups, process equipment,
fermenters, microisolator cages, and ventilated cage racks.
xxv
GLOSSARY
Primary
decontamination
technology
The initial validated equipment or process used to decontaminate

waste from the containment zone before disposal, incineration,
or release to sanitary sewers. This serves to remove or inactivate
infectious material or toxins through a process of disinfection,
sterilization, or inactivation. This may be followed by a secondary
decontamination process.
Prion
Small proteinaceous infectious particle generally considered to be

responsible for causing a group of neurodegenerative diseases
in humans and animals known as transmissible spongiform
encephalopathies.
Process equipment
Specic equipment used to carry out a manufacturing procedure

involving biological material. This term is generally used to
describe equipment used in large scale processes (e.g., industrial
fermentation equipment).
Program intent
A description of the planned work to be performed in a containment

zone. This includes, but is not limited to, the scope of work (e.g.,
diagnostic, teaching, research, large scale production, in vitro
work, in vivo work), a list of pathogens, toxins, and other regulated
infectious material to be handled or stored, a list of animal species
to be involved in in vivo work with pathogens and toxins in the
zone, and a list of procedures that may create aerosols.
Puff-back
The reversal of airow from the face of a Class II type B2

biological safety cabinet due to failure of the exhaust fan.
Release
The discharge of infectious material or toxins from a containment

system.
Representative
load
A simulation batch of materials of a similar type (e.g., gloves,

plastics, liquids) and quantity used to validate a decontamination
method for routine loads.
Restricted access
Access that is strictly controlled to authorized personnel only by

means of a physical barrier (i.e., a controlled access device or
system, such as an electronic access card, access code, etc.).
xxvi
Risk
The probability of an undesirable event (e.g., accident, incident,

breach of containment) occurring and the consequences of
that event.
Risk group (RG)
The classication of biological material based on its inherent

characteristics, including pathogenicity, virulence, risk of spread,
and availability of effective prophylactic or therapeutic treatments,
that describes the risk to the health of individuals and the public as
well as the health of animals and the animal population.
Risk management
plan
A plan that provides the foundation and organizational arrangements

for designing, implementing, monitoring, reviewing, and continually
improving risk management throughout the organization.
L\b^gmb\k^l^Zk\a
As dened in Section 1 of the Human Pathogens and Toxins

Regulations: the following types of systematic investigation or
research that are carried out in a eld of science or technology by
means of controlled activities:
a) basic research, when the controlled activities are conducted
for the advancement of scientic knowledge without a specic
practical application;
b) applied research, when the controlled activities are conducted
for the advancement of scientic knowledge with a specic
practical application;
c) experimental development, when the controlled activities are
conducted to achieve scientic or technological advancement
for the purpose of creating new or improving existing
materials, products, processes, or devices.
Sealable doors
Doors that are designed to allow leakage of air under normal

operating conditions yet are capable of being sealed to withstand
pressure decay testing and gaseous decontamination (e.g., threesided or four-sided gasket, four-sided door jamb).
Security barrier
A physical obstruction designed to prevent access to pathogens,

infectious material, toxins, or other related assets by unauthorized
personnel (e.g., locked doors, controlled access systems, or
padlocked storage equipment) that increases the security of a
containment zone by restricting access to authorized personnel only.
xxvii
GLOSSARY
Security sensitive
biological agents
(SSBAs)
The subset of human pathogens and toxins that have been

determined to pose an increased biosecurity risk due to their
potential for use as a biological weapon. SSBAs are identied
as prescribed human pathogens and toxins by Section 10 of the
Human Pathogens and Toxins Regulations. This means all Risk
Group 3 and Risk Group 4 human pathogens that are in the List
of Human and Animal Pathogens for Export Control, published
by the Australia Group, as amended from time to time, with the
exception of Duvenhage virus, Rabies virus and all other members
of the Lyssavirus genus, Vesicular stomatitis virus BOE
-ZNQIPDZUJD DIPSJPNFOJOHJUJT WJSVT; as well as all toxins listed in
Schedule 1 of the Human Pathogens and Toxins Act that are
listed on the List of Human and Animal Pathogens for Export
Control when in a quantity greater than that specified in
Section 10(2) of the Human Pathogens and Toxins Regulations.
Small-sized
animal
Refers to the physical size of the animal; small-sized animals

are small enough to be housed in primary containment caging.
Examples include rodents, rabbits, ferrets, chickens, and nonhuman primates. Small-sized animals may also be housed in an
animal cubicle (e.g., when open caging is used).
Small animal
containment zone
(SA zone)
Animal containment zone comprised of one or several co-located

or adjoining rooms of equal containment level where animals are
housed in animal rooms inside primary containment caging (e.g.,
microisolators). An SA zone may contain, for example, mice, rats,
rabbits, ferrets, or non-human primates, provided that they are
housed in primary containment caging.
Standard
operating
procedure (SOP)
A document that standardizes safe work practices and procedures

for activities with infectious material and toxins in a containment
zone, as determined by a local risk assessment.
Sterilization
Process that completely eliminates all living microorganisms,

including bacterial spores.
Strict animal
pathogen
A pathogen that causes disease exclusively in animals (i.e., not

capable of causing disease in humans).
xxviii
(Microbial) Toxin
A poisonous substance that is produced or derived from a

microorganism and can lead to adverse health effects in humans
or animals. Human toxins are listed in Schedule 1 and Part 1 of
Schedule 5 in the Human Pathogens and Toxins Act.
Terrestrial animal
pathogen
A pathogen that causes diseases in terrestrial animals, including

avian and amphibian animals, but excluding aquatic animals and
invertebrates.
Training needs
assessment
An evaluation performed to identify the current and future training

needs of the facility (organization, containment zone) and to
identify gaps in the current training program.
Transfer
A change in possession of pathogens, toxins, or other regulated

infectious material between individuals from the same or different
facilities (i.e., the movement from the place or places specied in
the licence or animal pathogen import permit to any other place).
Transportation
The act of transporting (e.g., shipping or conveyance) infectious

material or toxins to another building or location (i.e., different
address), within Canada or abroad, in accordance with the
Transportation of Dangerous Goods Act and Regulations.
Trigger quantity
The minimum quantity above which a toxin regulated by the

Human Pathogens and Toxins Act is considered a prescribed
toxin and, therefore, a security sensitive biological agent,
as described by Section 10(2) of the Human Pathogens and
Toxins Regulations.
Validation
The act of conrming that a method achieves its objective by

observing that specic parameters have been met (e.g., using
biological indicators to conrm that a given autoclave cycle can
decontaminate a representative load of waste). Validation infers
that a method is suitable for its intended purpose.
xxix
GLOSSARY
Ventilated cage
changing station
Equipment specically designed to change bedding and other

contents from animal cages that: a) directs the air away from the
user into the interior of the unit at a sufcient velocity to protect the
user from potential exposure to any infectious material or toxins;
and b) lters the exhaust air prior to release from the unit, thereby
preventing the potential release of infectious material or toxins into
the environment.
O^kb\Zmbhg
The routine monitoring of equipment and processes to ensure

continued efcacy between validations. This includes comparing
the accuracy of a piece of equipment to an applicable standard
or standard operating procedure (e.g., testing of a Class I
biological safety cabinet in accordance with the manufacturer
specications).
Virulence
The degree or severity of a disease caused by a pathogen.
Waste
Any solid or liquid material generated by a facility for disposal.
Zoonoses
Diseases that are transmissible between living animals and

humans. Zoonoses include anthropozoonoses (i.e., diseases
transmitted from animals to humans) and zooanthropoposes,
also known as reverse zoonoses (i.e., diseases transmitted from
humans to animals).
Zoonotic
pathogen
A pathogen that causes disease in humans and animals, and that

can be transmitted from animals to humans and vice versa (i.e.,
zoonoses). They are considered both human and animal pathogens.
xxx
NORMATIVE REFERENCES
NORMATIVE REFERENCES
American National Standards Institute (ANSI) / Sheet Metal and Air Conditioning
Contractors National Association, Inc. (SMACNA)
ANSI/SMACNA 016-2012, HVAC Air Duct Leakage Test Manual, 2nd Edition.
American Society of Mechanical Engineers (ASME)
ASME AG-1-2012, Code on Nuclear Air and Gas Treatment.
ASME N511-2007, In-service Testing of Nuclear Air Treatment, Heating,
Ventilating, and Air-Conditioning Systems.
Institute of Environmental Sciences and Technology (IEST)
IESTRP-CC001.5, HEPA and ULPA Filters.
IESTRP-CC00 5FTUJOH$MFBOSPPNT.
IESTRP-CC034, HEPA and ULPA Filter Leak Tests.
National Sanitation Foundation (NSF)/ American National Standards Institute (ANSI)
NSF/ANSI 49-201, Biosafety Cabinetry: Design, Construction, Performance,
and Field Certication.
National Standard of Canada (CAN)/ Canadian Standards Association (CSA)
CAN/CSA B64.10-11/B64.10.1-11, Selection and Installation of Backow
Preventers/Maintenance and Field Testing of Backow Preventers.
CAN/CSA Z180.1-13 (R2013), Compressed Breathing Air and Systems.
xxxii
CHAPTER 1
INTRODUCTION
CHAPTER 1 INTRODUCTION
1.1
Scope
The handling or storing of pathogens or toxins necessitates an awareness and application

of biosafety and biosecurity practices among personnel in laboratories and other
containment zones where work with pathogens, toxins, or infected animals is conducted.
The release of human and animal pathogens and toxins from laboratories or other
containment zones may pose a risk to public health, animal health, or both. Personnel can
minimize the risks associated with pathogens or toxins through the application of appropriate
biosafety and biocontainment principles and practices.
In Canada, facilities where human pathogens or toxins are handled or stored, such as
public health laboratories, teaching and research laboratories, diagnostic laboratories in
hospitals, and vaccine production plants, are regulated under the Human Pathogens and
Toxins Act (HPTA) and the Human Pathogens and Toxins Regulations (HPTR). Canadian
facilities that import animal pathogens, or animals, animal products or by-products (e.g.,
tissue, serum), or other substances that may carry an animal pathogen or parts thereof
are regulated by the Health of Animals Act (HAA) and the Health of Animals Regulations
(HAR). Zoonotic pathogens, capable of causing disease in human and animal hosts,
that are imported into Canada are regulated by the HPTA, HPTR, HAA, and HAR. The
Canadian Biosafety Standard (CBS), 2nd Edition, 2015 describes the minimum requirements
for containment zones in a regulated facility; this includes facilities that have been issued a
licence under the HPTA and HPTR, as well as facilities that have been issued an animal
pathogen import permit under the HAA and HAR. For facilities exempt from the licence
requirements under the HPTA and the HPTR and not importing under the HAA and the
HAR, it is considered best practice to demonstrate, by following the CBS requirements, that
all reasonable precautions have been taken to protect the health and safety of the public
against the risks associated with the materials in their possession.
The CBS sets out the physical containment requirements, operational practice
requirements, and i^k_hkfZg\^Zg]o^kb\Zmbhgm^lmbg`k^jnbk^f^gml for facilities where
human or terrestrial animal pathogens or toxins are handled or stored. Terrestrial animal
pathogens cause disease in terrestrial animals, including avian and amphibian animals,
but exclude pathogens of aquatic animals and invertebrates. Facilities in Canada where
imported aquatic animal pathogens are handled or stored must comply with the Canadian
Food Inspection Agencys (CFIAs) Containment Standards for Facilities Handling Aquatic
Animal Pathogens (CSFHAAP), 1st Edition, 2010. In situations where facilities handle
or store pathogens that cause disease in aquatic animals and either humans or terrestrial
animals, the facility is required to adhere to both the CBS as well as the CSFHAAP. Likewise,
facilities in Canada where imported plant pests are handled or stored must comply with
the CFIAs Containment Standards for Facilities Handling Plant Pests (CSFHPP), 1st Edition,
2007. Compliance with both the CBS as well as the CSFHPP is required in the event that an
imported plant pest is also capable of causing disease in either humans or terrestrial animals.
1.2
Regulatory Authorities
The Public Health Agency of Canada (PHAC) is the national authority on biosafety and
biosecurity for human pathogens and toxins. The PHAC is responsible for the regulation
of human pathogens and toxins under the authority of the HPTA and the HPTR and the
importation or transfer of pure cultures of terrestrial animal pathogens and toxins, with
the exception of non-indigenous animal pathogens and pathogens causing emerging
animal disease, under the authority of the HAA and the HAR.
The Canadian Food Inspection Agency (CFIA) is the national authority on biosafety
and biosecurity for foreign animal diseases and emerging animal diseases. The CFIA is
responsible for the regulation of the importation or transfer of pure cultures of non-indigenous
animal pathogens and emerging animal disease pathogens, as well as animals, animal
products, and animal by-products that contain a terrestrial animal pathogen, under the
authority of the HAA and HAR. In addition, the CFIA is responsible for the regulation of the
importation or transfer of aquatic animal pathogens and plant pests. Please contact the CFIA
for further details regarding requirements for these pathogens.
On December 1st, 2015, the HPTR comes into force and the CBS will come into effect and
supersede the Canadian Biosafety Standards and Guidelines (CBSG), 1st Edition, 2013.
The CBS will be used by the PHAC and the CFIA to verify the ongoing compliance of
regulated facilities. Compliance with the physical containment, operational practice, and
performance and verication testing requirements, respectively described in Chapters 3, 4,
and 5 of the CBS, will help prevent the inadvertent release of pathogens or toxins, which
could potentially pose signicant risks to the health of humans or animals, the environment,
or the economy.
In some instances, facilities may need to be upgraded or renovated to meet some of the
physical containment requirements outlined in Chapter 3. As per the current compliance
and enforcement program, the PHAC and the CFIA will review non-compliance items on a
case-by-case basis. Regulated parties are encouraged to discuss non-compliance items with
the relevant agency (or agencies) to determine a timeframe for compliance based on the
level of risk and the risk mitigation strategies in place or to determine if alternative mitigation
strategies can be implemented for these items.
For more information about the biosafety and biosecurity work of the PHAC, an e-mail
can be sent to 1)"$QBUIPHFOTQBUIPHFOFT"41$!DBOBEBDB, or visit the PHAC website
(www. publichealth.gc.ca/pathogens). For more information about the work of the CFIA,
an email can be sent to [email protected], or visit the CFIA website
(www.inspection.gc.ca/english/sci/bio/bioe.shtml).
1.3
Legislative and Regulatory Requirements

for Human Pathogens and Toxins
Under the HPTA and the HPTR, facilities conducting controlled activities involving human
pathogens or toxins have specic requirements in addition to the requirements described
in the CBS. A summary of these requirements is described in Tables 1-1 and 1-2 below;
this, however, is not an exhaustive list, and persons conducting controlled activities with
human pathogens or toxins are to refer to the specic sections of the HPTA or HPTR for
a complete understanding of the requirements. It remains the responsibility of the facility
(regulated party) to understand their obligations under both the HPTA and HPTR, in addition
to the applicable requirements set out in the CBS and their conditions of licence. The Human
Pathogens Importation Regulations will be repealed when the HPTR comes into effect, and
the licensing program outlined in the HPTR will replace the need for human pathogen and
toxin import permits.
Table 1-1. Summary of the key legislative and regulatory requirements for all facilities that
fall under the scope of the Human Pathogens and Toxins Act.
Key Requirements that apply to all facilities under the scope
of the HPTA (i.e., licensed and licence-exempted facilities)
Corresponding
Section(s) of HPTA/HPTR
Every person who knowingly conducts controlled activities

with human pathogens and toxins must take all reasonable
precautions to protect the health and safety of the public.
HPTA 6
Any activity with the human pathogens or toxins listed

in Schedule 5 of the HPTA is strictly prohibited.
HPTA 8
Unless otherwise exempted, a licence must be obtained from

the PHAC to authorize any of the following controlled activities:
s POSSESSING HANDLING OR USING A HUMAN PATHOGEN OR TOXIN
s PRODUCING A HUMAN PATHOGEN OR TOXIN
s STORING A HUMAN PATHOGEN OR TOXIN
s PERMITTING ANY PERSON ACCESS TO A HUMAN PATHOGEN OR TOXIN
s TRANSFERRING A HUMAN PATHOGEN OR TOXIN
s IMPORTING OR exporting a human pathogen or toxin;
s RELEASING OR OTHERWISE ABANDONING A HUMAN PATHOGEN
or toxin; or
s DISPOSING OF A HUMAN PATHOGEN OR TOXIN
HPTA 7(1)
Table 1-2. Summary of the legislative and regulatory requirements applicable only to
licence holders.
Key Requirements that apply to all licensed facilities
Corresponding
Licence Requirements
A licence applicant who intends to carry out l\b^gmb\

research must develop a plan that sets out how they
will administratively manage and control biosafety and
biosecurity risks during the term of the licence (i.e., risk
management plan), before the licence can be issued.
Licence applicants are required to designate a biological
lZ_^mrh_\^k (BSO) before a licence may be issued.
HPTR 3
HPTA 36(1)
Conditions of Licence
Licences are subject to conditions, and a licence holder and

all persons conducting controlled activities under a licence
must comply with those conditions.
HPTA 18(4)
HPTA 18(7)
Licence holders are required to communicate all licence

conditions to everyone conducting activities under that licence.
HPTA 18(6)
Corresponding
The following conditions apply to every licence:

1. licence holders and persons conducting controlled
activities authorized under the licence must not obstruct
a BSO when the BSO is exercising their powers or
carrying out their functions;
2. the BSO must be notied before arrangements are made
to do the following:
s IMPORT A HUMAN PATHOGEN OR TOXIN
s RECEIVE A HUMAN PATHOGEN OR TOXIN FROM ANOTHER FACILITY
s TRANSFER A HUMAN PATHOGEN OR TOXIN TO ANOTHER FACILITY
3. persons transferring human pathogens or toxins within
Canada must take reasonable care to satisfy themselves
that the intended recipient is licensed to work with the
agent or otherwise exempted from the requirement to
hold a licence;
4. persons exporting human pathogens and toxins
outside of Canada must take reasonable care to
satisfy themselves that the intended recipient will follow
applicable biosafety and biosecurity standards and
policies in the foreign jurisdiction;
5. intended recipients of human pathogens and toxins must
make reasonable efforts to locate human pathogens and
toxins that are not received within a reasonable time of
when they are expected to be received, and the BSO
must be notied of the situation without delay; and
6. persons who discover that they are inadvertently in
possession of a human pathogen or toxin that they are
not authorized to possess and that is not a Schedule
5 agent must immediately notify the BSO, ensure that
the human pathogen or toxin is handled and stored
appropriately, and dispose of it or transfer it to a licence
holder authorized to conduct controlled activities with that
pathogen or toxin within 30 days.
HPTR 4(1)
The following additional condition applies to every licence

that authorizes controlled activities with security sensitive
biological agents (SSBAs):
1. intended recipients of SSBAs who do not receive them
within 24 hours after the date and time they were expected
to be received must make reasonable efforts to locate the
SSBAs, inform the BSO without delay, and provide the
BSO with any relevant information.
HPTR 4(2)
Any licence conditions that are specic to a particular

licence are set out on the licence itself. Compliance with
the applicable requirements in the CBS will be included
as a licence-specic condition.
Corresponding
HPTA 18(4)
;bheh`b\ZeLZ_^mrH_\^kK^jnbk^f^gml
BSOs must have the following minimum qualications:

1. knowledge of microbiology appropriate to the risks
associated with the controlled activities being conducted,
attained through a combination of education, training,
and experience;
2. knowledge of the HPTA, the HPTR, and any applicable
federal or provincial legislation; and
3. knowledge of applicable biosafety and biosecurity
policies, standards, and practices appropriate to the risks
associated with the controlled activities being conducted.
HPTA 36(3)
HPTR 8
BSOs have the following functions:

1. verifying the accuracy and completeness of licence
applications;
2. communicating with the PHAC on behalf of the licence
holder;
3. promoting and monitoring compliance, including but
not limited to:
s ARRANGING FOR AND DOCUMENTING TRAINING RELATED
to biosafety and biosecurity policies, standards,
and practices;
s NOTIFYING THE 0(!# OF ANY INADVERTENT POSSESSION
of human pathogens and toxins or of any SSBAs
not received as expected;
s CONDUCTING PERIODIC INTERNAL INSPECTIONS AND BIOSAFETY
audits and reporting ndings to the licence holder;
s INFORMING THE LICENCE HOLDER OF ANY NON COMPLIANCE BY A
person conducting activities under the licence that is not
resolved after that person has been made aware of it;
4. assisting in the development and maintenance of the
Biosafety Manual and standard operating procedures
(SOPs) related to biosafety and biosecurity; and
5. assisting with internal investigations of incidents.
HPTA 36(5)
HPTR 9(1)
Corresponding
BSOs have the following power:

s MAY REQUIRE ANY PERSON WHO CONDUCTS CONTROLLED ACTIVITIES
under the licence to provide them with any records that
are necessary to assist them in carrying out their functions.
HPTA 36(5)
HPTR 9(2)
Human Pathogens and Toxins Act Security Clearance Requirements
Persons are prohibited from entering a part of a facility in

which controlled activities with SSBAs are authorized unless
they hold a valid Human Pathogens and Toxins Act
4ecurity $learance (HPTA 6ecurity &learance) issued by the
PHAC, or are accompanied and supervised by a person
who holds an HPTA 6ecurity &learance. This prohibition does
not apply if there are no SSBAs in that part of a facility, or if
any SSBAs that are present are locked up and inaccessible.
HPTA 33
HPTR 28
A holder of an HPTA 6ecurity &learance may only

accompany and supervise one person who does not hold a
clearance at any one time (1 to 1 ratio), and must be in the
same room and monitor their activities at all times.
HPTR 23
Persons who do not hold an HPTA 6ecurity &learance may

not be accompanied and supervised if their HPTA 6ecurity
&learance is suspended, or if they have previously been
refused an HPTA 6ecurity &learance or had one revoked
(and a new one has not been issued since that time).
HPTR 24
As part of the HPTA 6ecurity &learance application process,

applicants will need to submit a statement signed by a
licence holder certifying that they require an HPTA 6ecurity
&learance for a particular part or parts of a facility.
HPTR 12(1)(m)
The HPTA 6ecurity &learance is transferable between facilities,

but the holder of an HPTA 6ecurity &learance must provide a
new statement to the PHAC, signed by the licence holder,
certifying that they require an HPTA 6ecurity &learance before
an HPTA 6ecurity &learance holder enters a part of a facility that
was not authorized by their existing HPTA 6ecurity &learance.
HPTR 18
Corresponding
Reporting Requirements
The licence holder and the BSO must be notied whenever

the licence holder or any person who is conducting controlled
activities authorized under a licence intends to increase the
virulence, pathogenicity, communicability of a human
pathogen, or the resistance of a human pathogen to preventive
or therapeutic treatments, or to increase the toxicity of a toxin.
HPTR 5
Every person conducting controlled activities under authority of

a licence must inform the licence holder and the BSO without
delay of any inadvertent release or production of a human
pathogen or toxin, of any incident involving a human pathogen
or toxin that has caused, or may have caused, disease in an
individual, or if a pathogen or toxin has been stolen or is
otherwise missing.
HPTA 15
HPTR 4
Notication must be provided to the PHAC without delay

in the following circumstances:
s WHEN A LICENCE HOLDER HAS REASON TO BELIEVE THAT A HUMAN
pathogen or toxin has been released inadvertently from
a facility;
s WHEN A HUMAN PATHOGEN OR TOXIN THAT A PERSON IS
not authorized to possess is inadvertently produced
or otherwise comes into their possession;
s WHEN AN INCIDENT INVOLVING A HUMAN PATHOGEN OR TOXIN HAS
caused, or may have caused, disease in an individual;
s WHEN THERE IS REASON TO BELIEVE THAT A HUMAN PATHOGEN
or toxin has been stolen or is otherwise missing;
s WHEN A LICENCE HOLDER DECIDES TO PROHIBIT THE HOLDER
of an HPTA 6ecurity &learance from accessing a licensed
facility, including the reasons for that decision;
s WHEN THE DESIGNATED "3/ HAS CHANGED
s WHERE AN 33"! IS NOT RECEIVED WITHIN HOURS OF THE
date and time when it was expected to be received; and
s WHERETHEHOL DEROFAN(04! 6ECURITY &LEARANCEISCONVICTED
of a criminal offence.
HPTA 12(1)
HPTA 12(2);
HPTR 9(1)(c)(ii)
HPTA 13
HPTA 14
HPTA 32;
HPTR 7
HPTA 36(6)
HPTR 9(1)(c)(iii)
HPTR 19
Notication must be provided to the PHAC in the following

circumstances:
s before making any of the following changes, if they
could affect biocontainment, where controlled activities
with Risk Group 3 or 4 human pathogens or SSBA toxins
are conducted:
s changes to the physical structure of their facility;
s changes to any equipment; or
s changes to their SOPs; and
s within a reasonable time of a licence holder making
a name change.
Corresponding
HPTR 6(1)
HPTR 6(2)
Records and Documentation Requirements
Licence holders are required to keep a list of all persons

authorized to access a licensed facility, including persons
holding HPTA 6ecurity &learances and visitors.
HPTA 31
Licence holders are required to keep a record of the full

name of every person who enters, under accompaniment and
supervision, a facility where SSBAs are present, including the
date and the full name of the person who accompanied and
supervised them.
HPTR 25
10
1.4
Legislative and Regulatory Requirements for

Animal Pathogens, Toxins, and other Regulated
Infectious Material
Under the HAA and the HAR, Canadian facilities that import animal pathogens or part of
one that retains its pathogenicity (e.g., toxins), or animals, animal products (e.g., cream,
milk, eggs) or by-products (e.g., blood, serum, tissues), or other substances that may carry
an animal pathogen or part of one that retains its pathogenicity have specic requirements
in addition to the requirements described in the CBS. A summary of these requirements is
described in Table 1-3 below; this, however, is not an exhaustive list, and persons applying
for an animal pathogen import permit are to refer to the specic sections of the HAA or HAR
for a complete understanding of the requirements. It remains the responsibility of the facility
(importer) to understand their obligations under both the HAA and HAR, in addition to the
applicable requirements set out in the CBS and their conditions of import permit.
Table 1-3. Summary of the key legislative and regulatory requirements for importers under
the scope of the Health of Animals Act and the Health of Animals Regulations.
Key Requirements that apply to importers under the HAA
and HAR
Corresponding
Section(s) of HAA/HAR
The importation of any animal or other thing into Canada is

prohibited and subject to regulation under the HAR for the
purpose of preventing a disease or toxic substance from being
introduced into or spread within Canada.
HAA 14
No person shall possess or dispose of an animal or thing

that the person knows was imported in contravention of the
HAA or the regulations.
HAA 15(1)
An animal pathogen import permit must be obtained from the

PHAC or the CFIA prior to the importation of the following:
s AN ANIMAL PATHOGEN OR PART OF ONE THAT RETAINS ITS
pathogenicity (e.g., toxin); and
s AN ANIMAL ANIMAL PRODUCT ANIMAL BY PRODUCT OR OTHER
organism carrying an animal pathogen or part of one
that retains its pathogenicity (e.g., toxin).
HAR 51(a) and (b)
11
Key Requirements that apply to importers under the HAA

and HAR
An animal pathogen import permit may be issued by the PHAC

or the CFIA, provided that the relevant agency is satised that
the activity for which the permit is issued would not, or would
not be likely to, result in the introduction into Canada, the
introduction into another country from Canada, or the spread
within Canada of a vector, disease, or toxic substance.
Corresponding
Section(s) of HAA/HAR
HAR 160(1.1)
Conditions of Animal Pathogen Importation
Where any animal pathogen or part of one that retains its

pathogenicity (e.g., toxin) referred to in Section 51 is imported
into Canada under a permit, no person shall:
s MOVE THE IMPORTED PATHOGEN OR PART OF ONE THAT RETAINS
its pathogenicity (e.g., toxin) from the place or places
specied in the permit to any other place, except under
and in accordance with that permit or another permit
issued for that purpose; or
s INTRODUCE INTO AN ANIMAL THE IMPORTED PATHOGEN OR PART OF
one that retains its pathogenicity (e.g., toxin) except under
and in accordance with that permit or another permit
issued for that purpose.
Any animal pathogen import permit issued under the
HAR shall contain such conditions as the PHAC and/
or the CFIA consider advisable to prevent the introduction
of communicable disease into Canada or into any other
country from Canada and the spread of communicable
disease within Canada.
Every person to whom a permit is issued under the HAR shall
comply with the conditions contained in the animal pathogen
import permit.
12
HAR 51.1
HAR 160(1.1)(2)
HAR 160.1
CHAPTER 2
HOW TO USE THE

CANADIAN BIOSAFETY STANDARD
CHAPTER 2 HOW TO USE THE

CANADIAN BIOSAFETY STANDARD
The Canadian Biosafety Standard (CBS) describes, according to containment level,
the physical containment requirements, operational practice requirements, and
i^k_hkfZg\^ Zg] o^kb\Zmbhg m^lmbg` k^jnbk^f^gml for facilities where pathogens,
toxins, or other regulated infectious material are handled or stored. In the context of the
CBS, handling or storing pathogens or toxins includes possessing, handling, using,
producing, storing, permitting access to, transferring, importing, exporting, releasing,
disposing of, or abandoning such material.
2.1
Abbreviations, Denitions, and References
The CBS includes a detailed list of all abbreviations and acronyms used throughout; this
list is located at the beginning of the document. Each abbreviation or acronym is spelled
out upon rst use in a chapter and, in Chapters 3, 4, and 5, in each section or matrix,
with the abbreviation immediately following in brackets; the abbreviation is used exclusively
throughout the remainder of the chapter or matrix.
The CBS contains a comprehensive glossary of denitions for technical terms, located at the
beginning of the document; words dened in the glossary appear in bold type upon rst use
in each chapter and, in Chapters 3, 4, and 5, in each section or matrix. The terminology
used in the CBS is to be interpreted according to the corresponding denitions provided in
the glossary.
The CBS includes a list of normative references at the beginning of the document, providing
a summary of all external standards cited in the text. Except where information from an
external standard is directly incorporated into the CBS, external standards are referenced
by number only, and users are to refer to the most current version available. A full list of the
external standards and other documents that are referenced in the CBS is provided at the
end of the document.
2.2
Containment Levels and Containment Zones
Containment level refers to the minimum physical containment and operational practices
required for a containment zone where infectious material or toxins can be safely handled.
The CBS describes the three containment levels regulated by the Public Health Agency
of Canada (PHAC) and the Canadian Food Inspection Agency (CFIA), ranging from the
lowest level permitted to work with pathogens, toxins, and other regulated infectious material
(containment level 2 [CL2]) to the highest level of containment (containment level 4 [CL4]).
A containment zone itself is a physical area that meets the requirements for a specied
containment level. This can be a single room (e.g., a laboratory) or a series of co-located
rooms (e.g., several non-adjoining but lockable CL2 laboratory work areas), or it can be
comprised of several adjoining rooms of the same containment level (e.g., containment level
3 [CL3] suite comprised of dedicated laboratory work area and support areas, such as
14
anterooms, change rooms, storage rooms, preparation areas, wash up rooms, centralized
autoclave room). A containment zone may include one or more work areas of different types
(i.e., laboratory work area, large scale production area, animal work areas), as long as
they are of the same containment level.
2.3
Working with Human and Animal Pathogens

and Toxins
2.3.1
Pathogens and Risk Groups
A pathogen is a microorganism, nucleic acid, or protein capable of causing disease in

humans or terrestrial animals. This can include bacteria, viruses, fungi, parasites, prions,
recombinant DNA, genetically modied microorganisms, viral vectors, and synthetic biology
products. Human pathogens are capable of causing disease in humans; animal pathogens
cause disease in animals. For the purposes of the CBS, the term animal pathogens from
this point forward refers only to pathogens that cause disease in terrestrial animals, including
avian and amphibian animals. Zoonotic pathogens are pathogens that cause diseases
in humans and terrestrial animals and that can be transmitted from animals to humans or
vice versa (i.e., zoonoses), and are, therefore, considered as both human and animal
pathogens. In the context of the CBS, any isolate of a pathogen or any biological material
that contains human or animal pathogens is referred to as infectious material.
The following denitions provide the risk group categorization for both human and animal
pathogens based on the risk to the individual/animal and the risk to the community.
Examples of human pathogens are listed in Schedules 2 to 4 and in Part 2 of Schedule 5 of
the Human Pathogens and Toxins Act (HPTA), but these are not exhaustive lists. Examples of
animal pathogens can be found on the CFIA website.
2.3.1.1
Risk Group 1 (RG1; low individual and community risk)
A microorganism, nucleic acid, or protein that is either a) not capable of causing human
or animal disease; or b) capable of causing human or animal disease, but unlikely to
do so. RG1 organisms capable of causing disease are considered pathogens that pose
a low risk to the health of individuals or animals, and a low risk to public health and
the animal population. RG1 pathogens can be opportunistic and may pose a threat to
immunocompromised individuals. Neither of the RG1 subsets is regulated by the PHAC or
the CFIA due to the low risk to public health and the animal population.
2.3.1.2
Risk Group 2 (RG2; moderate individual risk, low community risk)
A pathogen or toxin that poses a moderate risk to the health of individuals or animals, and a
low risk to public health and the animal population. These pathogens are able to cause serious
disease in a human or animal but are unlikely to do so. Effective treatment and preventive
measures are available and the risk of spread of diseases caused by these pathogens is low.
Examples of RG2 human pathogens are included in Schedule 2 of the HPTA.
15
CHAPTER 2 HOW TO USE THE CANADIAN BIOSAFETY STANDARD
2.3.1.3
Risk Group 3 (RG3; high individual risk, low community risk)
A pathogen that poses a high risk to the health of individuals or animals, and a low risk to
public health. These pathogens are likely to cause serious disease in a human or animal.
Effective treatment and preventive measures are usually available and the risk of spread of
disease caused by these pathogens is low for the public. The risk of spread to the animal
population, however, can range from low to high depending on the pathogen. Examples of
RG3 human pathogens are included in Schedule 3 of the HPTA.
2.3.1.4
Risk Group 4 (RG4; high individual risk, high community risk)
A pathogen that poses a high risk to the health of individuals or animals and a high risk to
public health. These pathogens are likely to cause serious disease in a human or animal
which can often lead to death. Effective treatment and preventive measures are not usually
available and the risk of spread of disease caused by these pathogens is high for the public.
The risk of spread of disease to the animal population, however, ranges from low to high
depending on the pathogen. Examples of RG4 human pathogens are included in Schedule
4 of the HPTA.
2.3.2
Determining the Containment Level
In general, the containment level and risk group of the pathogen are the same (e.g., RG2
pathogens are handled at CL2); however, there are exceptions, and not all biological
material will fall perfectly into a given risk group or containment level following a risk
assessment. In some cases, there is a higher or unique level of risk associated when handling
certain pathogens (e.g., non-indigenous animal pathogens or prions) or with certain
types of work (e.g., in vivo work or in vitro work involving large scale volumes of pure
or concentrated cultures of pathogens). In other cases, biological material (e.g., tissues,
diagnostic specimens) may harbour pathogens, toxins, prions, or modied components of
a pathogen, in which case, a site-specic local risk assessment (LRA) is critical to evaluate
and determine which work-specic operational practices and mitigation strategies are to be
implemented to achieve the appropriate level of precaution.
It is very difcult to develop a comprehensive list of human and animal pathogens due to the
emergence of new pathogens and the ongoing research into the characteristics of existing
pathogens. Well-characterized pathogens that have had a pathogen risk assessment
completed by the PHAC or the CFIA have been assigned an appropriate risk group and
containment level. Risk assessments on well-characterized human and zoonotic pathogens
have been developed into technical documents by the PHAC known as Pathogen Safety Data
Sheets (PSDSs) that describe the hazardous properties of pathogens and recommendations for
their safe handling and are readily available on the PHAC website. Fact Sheets for federally
reportable diseases affecting terrestrial animals in Canada have been developed by the
CFIA and are also readily available on the CFIA website. The appropriate containment level
and any additional requirements to work with certain animal pathogens (e.g., non-indigenous
animal pathogens and emerging animal disease pathogens) are determined by the CFIA
through a pathogen risk assessment and containment level assessment.
16
2.3.3
Toxins
Biological toxins are poisonous substances that are a natural product of the metabolic
activities of certain microorganisms, plants, and animal species. Unlike pathogens, toxins
are non-infectious and unable to propagate when isolated from the parental organism. In the
context of the CBS, the word toxin refers only to microbial toxins regulated by the PHAC
and the CFIA under the HPTA and the Health of Animals Act (HAA). An exhaustive list of
toxins regulated under the HPTA is listed in Schedules 1 and 5 of the HPTA; whereas any
imported microbial toxin derived from an animal pathogen is also regulated under the HAA.
In general, toxins capable of producing human or animal disease are safely handled in CL2
zones, at a minimum. Additional physical containment or operational practice requirements
may be necessary, based on risk.
2.3.4
Prions
Prions are small, proteinaceous, infectious particles that are generally considered to be the
cause of a number of fatal progressive neurodegenerative diseases in humans and animals
known as transmissible spongiform encephalopathies. The most likely route of transmission of
infectious prions is through inoculation or ingestion. Prions are resistant to decontamination
procedures and processes commonly effective against other pathogens. Activities involving
infectious prions are generally assessed to be safely conducted at CL2 with specic
additional physical and operational requirements. Consequently, for certain requirements
presented in Chapters 3, 4, and 5, there are increased or unique requirements for activities
with prions indicated by the letter P in the matrix.
2.3.5
Security Sensitive Biological Agents
Security sensitive biological agents (SSBAs) are human pathogens and toxins that have
been determined to pose an increased biosecurity risk due to their inherent dual-use
potential for bioterrorism. In the context of the CBS, SSBAs are the human pathogens and
toxins that are identied as prescribed human pathogens and toxins in the HPTA and the
Human Pathogens and Toxins Regulations (HPTR). Prescribed human pathogens are all RG3
and RG4 human pathogens that also appear on the List of Human and Animal Pathogens
and Toxins for Export Control, published by the Australia Group (as amended from time
to time), except for Duvenhage virus, Rabies virus and all other members of the
Lyssavirus genus, Vesicular stomatitis virus BOE -ZNQIPDZUJD DIPSJPNFOJOHJUJT WJSVT.
Prescribed toxins are all toxins listed in Schedule 1 of the HPTA that also appear on
the List of Human and Animal Pathogens and Toxins for Export Control, published by
the Australia Group (as amended from time to time), when in a quantity greater than
the identified trigger quantity in a part of the facility where controlled activities with
SSBAs are authorized, as described by Section 10(2) of the HPTR. Consequently, for
certain requirements presented in Chapters 3, 4, and 5, there are unique requirements for
SSBAs at any containment level and, for activities with SSBA toxins in CL2 or CL2-Ag
zones, there are increased requirements (as indicated by the letter S in the CL2 or
CL2-Ag column of the matrix). A toxin present in a facility in a quantity below the
trigger quantity is not an SSBA; however, it remains a regulated toxin, and subject to
17
the requirements in the CBS (i.e., the minimum containment level for handling a regulated
toxin is CL2). For ease of reference, the PHAC maintains an exhaustive list (as amended
from time to time) of all SSBAs, including toxin trigger quantities, on the PHAC website
(http://phac-aspc.gc.ca/lab-bio/regul/ssba-abcse-eng.php).
2.3.6
Non-Indigenous Animal Pathogens
Non-indigenous animal pathogens are exotic to Canada (i.e., foreign animal disease agents
that are not present in Canada). In the context of the CBS, non-indigenous animal pathogens
are pathogens that are listed in the World OrganiTation for Animal Healths OIE-Listed
diseases, infections and infestations (as amended from time to time) and that are also exotic to
Canada. Due to the risk of serious negative health effects to the Canadian animal population
and the resultant economic impact, more stringent physical containment and operational
practice requirements are applied to containment zones where non-indigenous animal
pathogens are imported. Consequently, to prevent their release into the environment, there are
several unique requirements in Chapters 3, 4, and 5 for activities with non-indigenous animal
pathogens, as indicated in the text of the requirement itself. The CFIA continues to oversee the
facility certification program for facilities approved to handle or store imported non-indigenous
animal pathogens. Emerging animal disease pathogens are administered by the CFIA as nonindigenous animal pathogens due to the high risk of serious negative effects associated with
these pathogens. For ease of reference, example lists of non-indigenous animal pathogens
and emerging disease pathogens, sorted by risk group, are available through the Automated
Import Reference System (AIRS) on the CFIA website (http://www.inspection.gc.ca/airs).
2.3.7
Large Scale Work
The PHAC and the CFIA generally consider activities involving volumes of toxins or the
in vitro culture of infectious material on a scale of 10 litres or greater to be large scale;
this could be a single vessel with a volume of 10 litres or greater or, in some cases,
multiple vessels with a total volume of 10 litres or greater. Large scale production facilities,
such as industrial fermentation and vaccine production plants, pose an increased risk to
personnel and the environment due to the large quantities of infectious material or toxins
being handled. As such, there are sometimes more stringent requirements and additional
considerations when compared to laboratory work areas at the same containment level.
Chapters 3, 4, and 5 describe several additional requirements particular for containment
zones where activities with large scale volumes of infectious material or toxins occur, as
indicated in the text of the requirement itself. It will be determined in consultation with the
PHAC and the CFIA on a case-by-case basis whether or not particular activities conducted
in a containment zone are required to follow the increased or unique requirements for large
scale work areas.
18
2.3.8
Handling Risk Group 1 Biological Material
The legislation administered by the PHAC and the CFIA does not apply to RG1 human
and animal pathogens. The CBS does not specify the requirements for activities with RG1
material; however, due care should be exercised and safe work practices (e.g., good
microbiological laboratory practices) should be followed when handling these materials
since RG1 biological material does pose a low risk to the health of individuals or animals.
2.4
Working with Animals
Pathogen and toxin work performed in vivo involves living animals and is carried out in
an animal containment zone. An animal containment zone refers to a series of co-located
animal rooms or animal cubicles, as well as associated corridors and support rooms
(e.g., storage and preparation areas) of equal containment level. A zone where the animals
are contained in primary containment caging (i.e., ltered containment caging to prevent
the release of infectious material and toxins) is termed a small animal containment zone
(or SA zone). The room where animals are housed in primary containment caging within
an SA zone is referred to as an animal room. Alternatively, a zone where the room itself
provides the primary containment is termed a large animal containment zone (or LA
zone). The room or space within the LA zone in which animals are housed is referred to as
an animal cubicle. LA zones may also include animal post mortem rooms (PM rooms). In
the context of the CBS, the term post mortem room is specic to rooms in LA zones where
animal necropsies and dissections are conducted.
It is important to note that the designation as an SA zone or LA zone is dependent on the
way in which the animal is housed (primary containment caging versus the room providing
containment) rather than the actual physical size of the animal. In general, large-sized
animals and small-sized animals are housed in LA zones and SA zones, respectively.
However, in some cases, small-sized animals can be housed in an LA zone. For example,
a room where small-sized animals are housed in open caging only intended for the
connement of animals to an area (i.e., it does not include ltration to prevent the release
of infectious material and toxins) is considered to be an animal cubicle inside an LA zone,
despite the actual size of the animal. Animal cubicles and PM rooms in LA zones require
additional and sometimes unique physical containment and operational practice requirements
in order to contain the pathogens and toxins and protect personnel entering these spaces
from exposure. These requirements are identied under the agriculture (i.e., Ag) column.
19
2.5
Derogations and Exemptions to Physical

or Operational Requirements
As part of the ongoing pathogen risk assessments and containment level assessments
conducted by the PHAC and the CFIA, the containment level required for handling a pathogen
may change over time and specic physical containment or operational practice requirements
may be derogated (i.e., lessened) in conjunction with the implementation of additional risk
mitigation strategies to address the specic pathogen. For example, many of the physical
containment and operational practice requirements at CL3 are aimed at reducing the risks
associated with airborne or aerosol-transmitted pathogens; work with RG3 pathogens
not known to be transmissible by inhalation may allow certain physical or operational
requirements at CL3 to be derogated. The PHAC develops biosafety directives that provide
a comprehensive overview of the customized containment level requirements for activities with
specic pathogens, or groups of pathogens, when the containment level differs from the risk
group (e.g., RG3 pathogens handled in CL2). Derogations approved by the PHAC and/or
the CFIA would be stipulated in the licence or animal pathogen import permit.
Exemptions to specic physical or operational requirements will be considered by the PHAC
and the CFIA, provided that it can be demonstrated by the regulated party that the intent of
the requirement in question has been met through an alternative mechanism, as determined
by an LRA. All such exemptions will be evaluated on a case-by-case basis.
2.6
Matrix Layout: Chapters 3, 4, and 5
The requirements for facilities where pathogens, toxins, and other regulated infectious
material are handled or stored are provided in Chapters 3, 4, and 5. The requirements
are risk- and evidence-based, and, where possible, more performance-based than
explicitly prescriptive. Chapter 3 describes the physical containment requirements (i.e.,
engineering controls and facility design) that are to be met prior to the handling or storing
of infectious material or toxins. Chapter 4 describes the operational practice requirements
(i.e., administrative controls and procedures) to be implemented in order to mitigate risks
and protect personnel, the community, and the environment in relation to the handling of
infectious material or toxins. Chapter 5 provides the requirements for the performance
and o^kb\Zmbhg tests necessary to demonstrate compliance with the physical containment
requirements outlined in Chapter 3 and, in some cases, the operational practice
requirements outlined in Chapter 4.
The requirements in Chapters 3, 4, and 5 are presented in a series of matrices (or tables).
Each matrix has been laid out to illustrate which requirements apply to specic containment
levels and in some cases, to specic work areas. The requirements are grouped by topic into
multiple matrices that contain separate CL2, CL3, and CL4 columns. Rather than grouping
20
requirements by containment level within a given matrix, they are instead organized to
facilitate comparison and contrast of requirements related to a similar topic. This helps to
illustrate how the requirements at higher containment levels build on those of lower levels
of containment. Different types of work areas inside a containment zone are included in
each column of the matrices as summarized in Table 2-1. Since there are numerous
additional physical containment and operational practice requirements necessary for animal
containment zones where the room itself provides the primary containment, LA zones at
CL2 and CL3 are represented by separate columns, and are designated as CL2- and CL3Agriculture (i.e., CL2-Ag and CL3-Ag, respectively). The remaining work areas (laboratory
work areas, large scale production areas, and SA zones) are all represented under the CL2
and CL3 columns. The requirements in the CL4 column encompass all work areas.
Table 2-1. Summary of the types of work areas included in each column of the matrices.
Type of work area(s)
Laboratory work areas

Large scale production areas
SA zones (including animal rooms)
LA zones (including animal cubicles
and PM rooms, where applicable)
*
CL2*
CL2Ag*
3
3
3
CL3
CL3Ag
3
3
3
3
CL4
3
3
3
3
includes activities involving prions or animals infected with prions

animal containment zones where the animals are housed in primary containment caging
animal containment zones where the room itself provides the primary containment
In most cases, the requirements for all work areas within a column of the matrices are the
same (e.g., CL2 laboratory work area, large scale production area, and SA zones). In
some instances, the requirement itself will reference a particular type of work area (e.g.,
large scale, animal work, SA zone, animal rooms, animal cubicles, PM rooms)
to which it applies and a closed box will appear in the columns ( ). In the case that a
requirement applies to CL2 SA zones and CL2 large scale zones, but is not required for
CL2 laboratory work areas, an open box ( ) appears in the CL2 column.
In instances where the requirements differ for different work areas, the exceptions are
listed in square brackets below the requirement (e.g., [Not required for CL2 SA zones.]).
The absence of a symbol in a column indicates that the element is not required for that
containment level. The description of the symbols and an example of the matrix layout is
provided in Figure 2-1.
21
Throughout Chapters 3, 4, and 5, the following symbols and combinations are used:
Required for all containment zones, including work areas where
activities with prions and SSBAs are conducted
For CL2 and CL2-Ag zones, the following symbols are used to facilitate the identication
of the applicable requirements.
Required for all CL2 SA zones and CL2 large scale production areas
including CL2 SA zones and CL2 large scale production areas where
activities with prions or SSBAs are conducted; not required, for CL2
laboratory work areas
P
Required for all work areas where activities with prions are conducted
Required for all work areas where activities with SSBAs are conducted
PS
Required for all work areas where activities with prions or SSBAs
are conducted
Required for all CL2 SA zones and CL2 large scale production areas,
as well as for CL2 laboratory work areas where activities with prions
are conducted
PS
Required for all CL2 SA zones and CL2 large scale production areas,
as well as for CL2 laboratory work areas where activities with prions
or SSBAs are conducted
22
Figure 2-1: Example of the matrix layout and the symbols found in Chapters 3, 4, and 5.
matrix # requirement #
3.2
3.2.1
containment level (CL)
matrix title
Containment Barrier
Openable windows positioned on the

containment barrier to include effective
pest control and security.
CL2
CL3
CL3Ag
CL4
} requirement
3.2.2
Windows on the containment barrier to

be closed and secured at all times.
3.2.3

be non-opening and sealed.
3.2.10
All penetrations of the containment

barrier of the containment zone, animal
cubicle, and PM room at or below the
work surface and any other surface that
may become contaminated, including all
conduits and wiring, to be sealed with a
non-shrinking sealant that is compatible
with the disinfectant(s) in use.
[Only applies to CL2 laboratory

work areas.]
Exemptions to the requirement are listed

in square brackets below the requirement.
CL2Ag
Required only for CL2

areas where prions or
SSBAs are handled.
PS
Requirement for all

CL2 activities in SA
and large scale zones.
P
Required for CL2 areas

(laboratory work areas,
large scale production areas,
and SA zones) where prions
are handled.
23
2.7
Explanatory Notes
The Explanatory Notes, found in the Appendix, provide additional information to identify
the risk(s) that are being mitigated by each requirement listed in Chapters 3, 4, and 5. The
Explanatory Notes also provide examples of how the requirement can be achieved. The
Explanatory Notes for the requirements listed in Chapter 5 provide more details on the tests
to be performed. The Explanatory Notes are organized in such a way that each requirement
in Chapters 3, 4, and 5 has a corresponding entry in the Explanatory Notes, following
the same numbering convention. It is important to note that the examples included in the
Explanatory Notes are not provided as specic requirements or recommendations; rather
they are provided for clarication and represent typical means of meeting a requirement.
2.8
The Canadian Biosafety Handbook
The Canadian Biosafety Handbook (CBH), 2nd Edition, 2015, is a companion document to
the CBS that provides the core information and guidance on how the biosafety requirements
outlined in the CBS can be achieved. The second edition of the CBH provides an update
of the Guidelines provided in Part II of the Canadian Biosafety Standards and Guidelines
(CBSG), 1st Edition, 2013. The CBH is structured to systematically address the concepts
required for the development of a comprehensive risk-based biosafety management
program. The CBH provides general guidance for containment zone personnel rather than
specic guidance or standard operating procedures (SOPs) for individual pathogens.
24
CHAPTER 3
PHYSICAL CONTAINMENT
REQUIREMENTS
CHAPTER 3 PHYSICAL CONTAINMENT

REQUIREMENTS
This chapter describes the physical containment requirements designed to mitigate the
risks associated with handling or storing pathogens, toxins, infected animals, or other
regulated infectious material. Physical containment is achieved through specic physical
barriers provided by engineering controls and facility design. The phrase in accordance
with function is included in some requirements where the specic activities and procedures
performed in the containment zone (i.e., the function) may inuence how the requirement
is implemented. Details on the use and interpretation of the matrices that follow are provided
in Chapter 2. A description of the symbols used appears in Section 2.6.
3.1
Structure and Location
The site selection process for a containment zone generally includes an assessment of
local programs and the local environment. Consideration of the risks, including the impact
of possible pathogen or toxin release, is important at the beginning of the design phase
and before construction work begins. In areas prone to natural disasters, buildings and
support systems for containment zones may need to meet more stringent building codes.
3.1
Structure and Location
3.1.1
Containment zones to be separated

from public and administrative areas
by a door.
3.1.2
Dedicated paper/computer work

stations within the containment zone to
be segregated from laboratory work
stations, animal rooms, animal cubicles,
and post mortem rooms (PM rooms).
3.1.3
Structure and location of containment

zone to be designed to withstand internal
and external environmental factors.
3.1.4
Laboratory work areas to be located

outside of animal cubicles.
3.1.5
Cold storage area or equipment to be

provided in, or adjacent to, the PM room.
26
CL2
CL2Ag
CL3
CL3Ag
CL4
3.2
Containment Barrier
The containment barrier refers to the physical structure(s) or obstruction(s) present that
create a boundary between the clean and dirty areas of a containment zone. The
containment barrier itself is created by the walls, doors, oors, and ceilings of a room
that physically enclose the areas where infectious material, toxins, and/or infected
animals are handled or stored. In containment zones where bgpZk] ]bk^\mbhgZe Zbkhp
(IDA) is provided, the containment barrier is also maintained through negative air pressure
differentials and IDA. Points of access through the containment barrier are provided through
doors and anterooms. Equipment such as dunk tanks, pass-through chambers, and
double-door barrier autoclaves, are examples of penetrations of the containment barrier.
3.2
3.2.1
Containment Barrier
CL2
CL2Ag
CL3
CL3Ag
Openable windows positioned on the

containment barrier to include effective
pest control and security.
[Only applies to CL2 laboratory
work areas.]
3.2.2
Windows on the containment barrier

to be closed and secured at all times.
3.2.3
Windows on the containment barrier

to be non-opening and sealed.
3.2.4
Window glazing material to provide the

appropriate level of security as determined
by a biosecurity risk assessment.
3.2.5
PS
PS

be positioned to prevent public viewing
into animal rooms, animal cubicles,
and post mortem rooms (PM rooms).
[Not required for CL2 large scale
production areas.]
3.2.6
Equipment located on the containment

barrier to be designed and installed in a
manner that maintains the integrity of the
containment barrier.
27
CL4
CHAPTER 3 PHYSICAL CONTAINMENT REQUIREMENTS
3.2
Containment Barrier
3.2.7
Pass-through technologies on the

containment barrier to be equipped with
doors that have mechanical or electronic
interlocks (preferred), or visual/audible
alarms, or other acceptable mechanism,
that prevent the simultaneous opening
of both doors.
3.2.8
Pass-through technologies on the

containment barrier to be equipped with
doors that have mechanical or electronic
interlocks that prevent the simultaneous
opening of both doors.
3.2.9
Doors of pass-through technologies on

the containment barrier to be equipped
with mechanical or electronic interlocks
and visual/audible alarms that prevent
the simultaneous opening of both doors.
3.2.10
All penetrations of the containment

barrier of the containment zone, animal
cubicle, and PM room at or below the
work surface and any other surface that
may become contaminated, including all
conduits and wiring, to be sealed with
a non-shrinking sealant that is compatible
with the disinfectant(s) in use.
3.2.11
All penetrations of the containment barrier,

including all conduits and wiring, to be
sealed with a non-shrinking sealant that is
compatible with the disinfectant(s) in use.
3.2.12
All penetrations of the containment barrier

inside animal cubicles and PM rooms,
including all conduits and wiring, to be
sealed with a non-shrinking sealant that is
compatible with the disinfectant(s) in use.
28
CL2
CL2Ag
CL3
CL3Ag
CL4
3.3
Access
Physical and security barriers (e.g., doors, locks, anterooms, interlocks) at points of
entry into and exit from the containment zone are critical to maintaining containment
integrity and allowing only trained and authorized individuals access to the zone. In high
containment zones, the physical barriers help maintain bgpZk]]bk^\mbhgZeZbkhp (IDA)
and provide space so that contaminated or potentially contaminated personal protective
equipment (PPE) remains inside the containment barrier.
3.3
Access
3.3.1
Doors to the containment zone

to be lockable.
3.3.2
Biohazard warning signage (including

the international biohazard warning
symbol, containment level, name and
telephone number[s] of contact person,
and entry requirements) to be posted at
the containment zone point(s) of entry.
3.3.3
Where unique hazards exist, projectspecic signage to be posted at the

animal room, animal cubicle, and post
mortem room (PM room) point(s) of entry.
3.3.4
Restricted access into the part of

the facility where security sensitive
biological agents (SSBAs) are stored
and handled to be provided through
a controlled access system.
3.3.5
Restricted access into the containment

zone to be provided through a controlled
access system.
3.3.6
Restricted access into each animal room,

animal cubicle, and PM room to be
provided through a controlled access
system or other acceptable mechanism.
CL2
CL2Ag
CL3
CL3Ag
29
CL4
3.3
Access
3.3.7
Non-reproducible keys to be used when

key-locks are used as the controlled
access system.
3.3.8
Electronic controlled access system to the

containment zone, where provided, to be
backed up with an alternate controlled
access system or other acceptable means.
3.3.9
Space to be provided for the storage

of PPE in use.
3.3.10
Dedicated change area to be provided at

personnel entry to the containment zone to
allow for separation of personal clothing
from dedicated containment zone clothing
(i.e., clean change area separated
from dirty change area).
3.3.11
Anteroom(s) to be provided at the

point(s) of entry into/exit from the
containment zone.
3.3.12
Anteroom(s) to be provided at the point(s)

of entry into/exit from:
s THE CONTAINMENT ZONE or
s EACH ANIMAL CUBICLE AND 0- ROOM
3.3.13
Anteroom(s) to be provided for entry

into/exit from:
s THE CONTAINMENT ZONE and
s EACH ANIMAL CUBICLE AND 0- ROOM
(except entry/exit through the dirty
corridor).
30
CL2
PS
CL2Ag
CL3
CL3Ag
CL4
3.3
Access
3.3.14
Anteroom(s) at the point(s) of exit from the

containment zone to include:
s A WALK THROUGH BODY SHOWER LOCATED
on the containment barrier between
the dirty and clean change areas.
3.3.15
Anteroom(s) at the point(s) of exit from

the containment zone and each animal
cubicle and PM room (except exit to the
dirty corridor) to include:
3.3.16
Anteroom(s) at the point(s) of exit from the

containment zone to include:
s A CHEMICAL decontamination shower
and a suit change area (may also be
dirty change area), in containment
zones where positive-pressure suits
are worn; and
3.3.17
Anteroom critical door(s) to be

provided with a physical or operational
mechanism that prevents the simultaneous
opening with:
s THE DOOR LEADING INTO THE ANTEROOM
from outside of the containment zone;
and
s THE DOORS LEADING FROM THE ANTEROOM
into the laboratory work area/
animal room/animal cubicle/
PM room.
CL2
CL2Ag
CL3
CL3Ag
31
CL4
3.3
3.3.18
3.3.19
Access
Anteroom critical door(s) to be provided

with mechanical or electronic interlocks
(equipped with manual overrides
for emergency exit) that prevent the
simultaneous opening with:
s THE DOOR LEADING INTO THE ANTEROOM FROM
outside of the containment zone; and
s THE DOORS LEADING FROM THE ANTEROOM
into the laboratory work area/animal
room/animal cubicle/PM room.
Chemical decontamination shower
doors to be provided with mechanical or
electronic interlocks (equipped with manual
overrides for emergency exit) that prevent
the simultaneous opening of doors.
[Not required in laboratory work areas
where infectious material is exclusively
handled in a Class III biological safety
cabinet (BSC) line.]
3.3.20
Sealable doors to be provided at

the point(s) of animal entry into animal
cubicles (i.e., between the cubicle and
corridor) and PM rooms (i.e., between
PM room and corridor), except doors
to the dirty corridor.
32
CL2
CL2Ag
CL3
CL3Ag
CL4
3.3
3.3.21
Access
CL2
CL2Ag
CL3
CL3Ag
Airtight doors to be provided for

entry, including:
s THE CHEMICAL DECONTAMINATION
shower doors;
s THE INNER AND OUTER DOORS OF THE
anteroom(s) dedicated to the entry
of animals and equipment into
the containment zone; and
s ANY CRITICAL DOOR DIRECTLY ON
the containment barrier.
[Not required for CL3-Ag zones where
only human and/or indigenous animal
pathogens are handled or CL4 laboratory
work areas where infectious material is
exclusively handled in a Class III BSC line.]
33
CL4
3.4
Surface Finishes and Casework
Selecting the appropriate surface nishes and casework for containment zones is necessary
to facilitate the maintenance, cleaning, and decontamination of surfaces within the zone.
Surface nishes also help protect against the stresses associated with activities routinely
performed within the containment zone, such as repeated decontamination and frequent
high pressure washing in animal containment zones.
3.4
3.4.1
Surfaces and interior coatings, including,

but not limited to, oors, ceilings, walls,
doors, frames, casework, benchtops, and
furniture, to be cleanable, non-absorbent,
and resistant to scratches, stains, moisture,
chemicals, heat, impact, repeated
decontamination, and high pressure
washing, in accordance with function.
3.4.2
Surfaces to be continuous with adjacent

and overlapping materials.
3.4.3
Benches, doors, drawers, handles, and

shelving to have smooth rims and corners
where positive-pressure suits are worn.
3.4.4
Backsplashes, when installed tight to

the wall, to be sealed at the wall-bench
junction and continuous with work surfaces.
3.4.5
Floors to be slip-resistant in accordance

with function.
3.4.6
Floors in animal rooms, animal

cubicles, post mortem rooms
(PM rooms), and corridors to withstand
loading, in accordance with function.
production areas.]
34
CL2
CL2Ag
CL3
CL3Ag
CL4
3.4
3.4.7
Continuity of seal to be maintained

between the oor and wall.
3.4.8
Continuity of seal to be maintained

between the wall and ceiling.
3.4.9
Interior surface materials to restrict

penetration of gases and liquids used
for decontamination and/or laboratory
purposes.
3.4.10
Protruding obstructions to be minimized

and appropriately shielded in animal
cubicles and corridors.
CL2
CL2Ag
CL3
CL3Ag
35
CL4
3.5
Air Handling
The heating, ventilation, and air conditioning (HVAC) systems can be designed to create a
dened containment barrier to minimize the spread of infectious aerosols or aerosolized
toxins. These systems, particularly in high containment zones, incorporate secondary
containment barriers such as bgpZk] ]bk^\mbhgZe Zbkhp (IDA) and abà ^_\b^g\r
iZkmb\neZm^Zbk!A>I:"em^kl for exhaust air.
3.5
3.5.1
Air Handling
IDA to be provided where:

s pathogens that are primarily
infectious through inhalation are
handled; or
s INFECTIOUS AEROSOLS OR AEROSOLIZED
toxins may be generated by
procedures in use.
[Not required for SA zones.]
3.5.2
IDA to be provided and maintained under

normal operation of the HVAC system.
3.5.3
Monitoring device(s) that visually

demonstrate IDA to be provided
for the containment zone.
3.5.4
Pressure differential monitoring lines

penetrating the containment barrier
to be provided with HEPA ltration
or acceptable alternative.
[Not required for CL3 zones with airtight
pressure differential monitoring devices.]
3.5.5
Audible or visual alarms that signal HVAC

systems failure to be provided to alert
containment zone personnel inside and
outside the containment zone.
36
CL2
CL2Ag
CL3
CL3Ag
CL4
3.5
Air Handling
3.5.6
Supply and exhaust air systems to be

independent of other areas. CL3 air
systems may be combined with areas of
lower containment when provided with
effective backdraft protection.
3.5.7
Supply air duct to areas inside the

containment barrier to be provided with
effective backdraft protection capable
of isolation from inside or outside of the
3.5.8
Supply air to be HEPA ltered.
3.5.9
Supply and exhaust air systems to be

provided with automatic mechanical/
electronic interlocks that prevent
sustained positive pressurization of
the containment zone.
3.5.10
CL2
CL2Ag
CL3
CL3Ag
Where IDA is provided, exhaust air to be:

s PASSED THROUGH A lLTER THAT PREVENTS
the release of infectious material
or toxins; or
s EXHAUSTED DIRECTLY TO
the outdoors.
[Not required for SA zones.]
3.5.11
Exhaust air to be passed through

HEPA ltration.
37
CL4
3.5
Air Handling
3.5.12
Exhaust air to be passed through

two stages of HEPA ltration.
3.5.13
HEPA lters to conform to

IEST-RP-CC001.5.
3.5.14
HEPA lter housings to be designed to

withstand structural changes at applied
pressure of 1000 Pa (i.e., 4 inches water
gauge [in. w.g.]) in accordance with
ASME N511 and AG-1.
3.5.15
HEPA lter housings to be designed

to allow in situ lter isolation,
decontamination, and testing.
3.5.16
Supply air ductwork located between

the containment barrier and backdraft
protection to be sealed airtight in
accordance with ANSI/SMACNA
016 Seal Class A.
3.5.17
Exhaust air ductwork located between the

containment barrier and HEPA lter(s) or
isolation damper(s) to be sealed airtight
in accordance with ANSI/SMACNA
016 Seal Class A.
3.5.18
Effective airow control devices to

be provided on supply and exhaust
air systems.
3.5.19
Sections of supply and exhaust air systems

located outside the containment barrier to
be accessible for maintenance and repair.
38
CL2
CL2Ag
CL3
CL3Ag
CL4
3.6
Facility Services
Facility services include all plumbing, electrical, and other services related to the operation
of the containment zone.
3.6
Facility Services
3.6.1
Exposed conduits, piping, and other

services to be mounted to allow for
decontamination of all surfaces.
3.6.2
Individual and/or dedicated main water

supply shut-off valves and other controls
to be located and accessible from
3.6.3
Water supply services to be provided

with isolation valve and [Z\dhp
prevention in accordance with CAN/
CSA-B64.10/B64.10.1.
3.6.4
CL2
CL2Ag
CL3
CL3Ag
Sinks to be provided and located to

facilitate handwashing upon exit from
[Not required for CL4 zones where
positive-pressure suits are worn.]
3.6.5
3.6.6
Sinks provided for handwashing to be

equipped with hands-free capability.
Emergency eyewash and shower
equipment to be provided in accordance
with containment zone activities.
39
CL4
3.6
3.6.7
Facility Services
CL2
CL2Ag
Containment zone to be designed to

control the release of large scale
process uids into sanitary sewers.
[Not required for CL2 SA zones.]
3.6.8
Containment zone to be designed to

contain the release of the full volume
of large scale process uids.
3.6.9
Drains to be equipped with deep seal

traps of sufcient depth to maintain water
seal and to prevent suction of liquid
wastes back into containment zone.
3.6.10
Containment zone drain piping servicing

areas inside the containment barrier to
be independent from those of other areas
and directly connected to an ^_n^gm
decontamination system.
[In CL3 zones, only required where
non-indigenous animal pathogens
are handled.]
3.6.11
Autoclave condensate drains located

outside the containment barrier to
have a closed connection and be
directly connected to the drain piping
servicing areas inside the containment
barrier, unless condensate is effectively
decontaminated prior to release.
3.6.12
Plumbing vent lines to be independent

from those of lower containment, unless
provided with abà^_\b^g\riZkmb\neZm^
Zbk!A>I:"emkZmbhg upstream from the
connection.
40
CL3
CL3Ag
CL4
3.6
3.6.13
Facility Services
CL2
CL2Ag
CL3
CL3Ag
Plumbing vent lines to be provided with

HEPA ltration with a means of isolation
and decontamination.
are handled.]
3.6.14
Plumbing vent lines to be independent

from those of lower containment and
provided with two stages of HEPA
ltration with a means of isolation
and decontamination.
3.6.15
Supplied breathing air and air hose

connections to be provided in areas
where positive-pressure suits are worn.
3.6.16
Backup air supply system that allows

sufcient time for emergency evacuation
to be provided in areas where positivepressure suits are worn.
3.6.17
Light ballasts and starters to be located

outside the containment barrier.
3.6.18
Services and equipment critical to

maintaining containment and biosecurity
to be supported by emergency power.
3.6.19
Life safety systems, building automation

systems, and security systems to be
supported by uninterruptJCMe power
supply (UPS).
PS
41
CL4
3.7
Essential Biosafety Equipment
Essential biosafety equipment is key to ensuring effective containment of pathogens,

toxins, and other regulated infectious material. This includes all primary containment
devices (e.g., biological safety cabinets [BSCs], isolators, centrifuges with sealable
cups, process equipment, fermenters, microisolator cages, ventilated cage racks, sealed
biological waste containers).
3.7
3.7.1
Certied BSCs and other primary

containment devices to be provided,
based on work activities.
3.7.2
Certied BSCs and other primary

containment devices to be provided.
3.7.3
Class II B2 BSCs, where present, to

be installed and set-up in a manner to
eliminate reversal of airow from the
face of the BSC (i.e., puff-back) during
a failure of the heating, ventilation, and
air conditioning (HVAC) system or the
BSC exhaust fan; where elimination of
puff-back cannot be achieved, the risk
associated with puff-back to be mitigated
through physical and operational means.
3.7.4
Process equipment, closed systems,

and other primary containment devices
to be designed to prevent the release
of infectious material or toxins.
3.7.5
Process equipment for large scale

activities with infectious material or toxins
to be equipped with sensing devices
to monitor containment integrity during
operations and to signal failure.
42
CL2
CL2Ag
CL3
CL3Ag
CL4
3.7
3.7.6
3.7.7
CL2
CL2Ag
CL3
CL3Ag
BSCs, where present, to be located as

far as possible from high trafc areas,
doors, openable windows, and air
supply/exhaust diffusers.
Large reusable equipment for large scale
activities with infectious material or toxins
to be designed and constructed to be
effectively cleaned and decontaminated,
or sterilized, in a manner that reduces
personnel exposure.
3.7.8
3.7.9
Primary containment caging to be

provided to house infected animals.
[Not required for large scale
production areas.]
Abà^_\b^g\riZkmb\neZm^Zbk!A>I:"&
em^k^] primary containment caging
systems, or partial containment caging
systems that are housed in HEPA-ltered
ventilated enclosures, to be provided
to house infected animals.
[Not required for laboratory work areas
or large scale production areas.]
43
CL4
3.7
3.7.10
Animal cages and animal cubicles to

be designed to prevent animal escape.
production areas.]
3.7.11
Decontamination technologies for

the decontamination of materials to be
provided within the containment zone,
or standard operating procedures
(SOPs) to be in place to safely and
securely move or transport waste out
of the containment zone to a designated
decontamination area.
3.7.12
Decontamination technologies for

the decontamination of contaminated
materials to be provided inside the
3.7.13
Decontamination technologies to be
provided on the containment barrier.
3.7.14
Decontamination technologies to be
provided with monitoring and recording
devices that capture operational
parameters.
3.7.15
An autoclave, where present, to be

capable of operating at the appropriate
temperature for decontamination,
as determined by validation.
44
CL2
CL2Ag
CL3
CL3Ag
CL4
3.7
3.7.16
Supply tanks for chemical decontamination

technologies to be equipped with alarms
that signal low levels.
3.7.17
Vacuum systems to be equipped with

a mechanism that prevents internal
contamination.
3.7.18
Two-way communication system(s) to be

provided inside the containment barrier
that allows communication between
inside the containment barrier to outside
the containment zone, in accordance
with function.
3.7.19
Observation windows and/or video

equipment to be installed in a manner that
allows activities to be visually monitored
from outside the containment barrier.
CL2
CL2Ag
CL3
CL3Ag
45
CL4
3.8
Efuent Decontamination Systems
>_n^gm ]^\hgmZfbgZmbhg lrlm^fl prevent the release of contaminated liquids into

sanitary sewers, and ultimately, the environment. An efuent decontamination system is
critical for decontaminating all liquid waste generated in CL3 zones where non-indigenous
animal pathogens are handled, large animal containment zones (LA zones) where
prions are handled, and all CL3-Ag and CL4 zones.
3.8
3.8.1
>_n^gm=^\hgmZfbgZmbhgLrlm^fl
An efuent decontamination system

appropriate to the work being conducted
to be provided.
are handled.]
CL2
CL2Ag
3.8.2
Efuent decontamination system to be

capable of operating at the appropriate
parameters for decontamination,
as determined by validation.
3.8.3
Drain piping connected to an efuent

decontamination system to be sloped
to ensure gravity ow.
3.8.4
Efuent decontamination system to be

heat and chemical resistant consistent
with use.
46
CL3
CL3Ag
CL4
3.8
3.8.5
Rooms housing an efuent decontamination

system serving as a primary
decontamination technology to:
s HAVE LOCKABLE DOORS
s HAVE DOORS WITH BIOHAZARD
warning signage;
s HAVE SEALED mOOR SURFACES
s HAVE mOOR DRAINS THAT ARE SEALED
or re-routed to the efuent
decontamination system; and
s HAVE SUITABLE personal protective
equipment (PPE) and a spill kit
available for emergency response.
3.8.6
Rooms housing an efuent decontamination

system serving as a primary
decontamination technology to:
s HAVE LOCKABLE DOORS
s HAVE DOORS WITH BIOHAZARD
warning signage;
s CONTAIN THE RELEASE OF THE FULL
volume of the largest holding tank
operating capacity of the efuent
decontamination system;
s HAVE SEALED mOOR SURFACES
s HAVE mOOR DRAINS THAT ARE SEALED
or re-routed to the efuent
decontamination system;
s MAINTAIN inward directional
Zbkhp (IDA);
s HAVE AN anteroom for entry/exit;
s HAVE abà^_\b^g\riZkmb\neZm^Zbk
!A>I:"emkZmbhg on exhaust air; and
s HAVE SUITABLE 00% AND A SPILL KIT
available for emergency response.
CL2
CL2Ag
CL3
CL3Ag
47
CL4
3.8
CL2
CL2Ag
3.8.7
Efuent decontamination system to include

a mechanism that prevents blockages.
3.8.8
Alarm system to be provided to

indicate warnings and failure of efuent
decontamination system.
3.8.9
Thermally controlled efuent

decontamination systems to be equipped
with temperature monitoring devices that
are calibrated annually.
3.8.10
Drain piping connected to an efuent

decontamination system to be identied
with labels.
3.8.11
Drain piping leading to an efuent

decontamination system to be accessible
for maintenance and repair.
3.8.12
Efuent decontamination system vent lines

to be provided with HEPA ltration.
3.8.13
Efuent decontamination system vent

lines to be provided with two stages
of HEPA ltration.
48
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CHAPTER 4
OPERATIONAL PRACTICE
REQUIREMENTS
CHAPTER 4 OPERATIONAL PRACTICE

REQUIREMENTS
This chapter describes the operational practice requirements designed to mitigate risks
associated with handling or storing pathogens, toxins, or other regulated infectious
material, including infected animals. Operational practice requirements are achieved
through specic administrative controls and by performing specic documented procedures.
Although the requirements in this chapter are specied for each containment zone,
institutions or organizations may decide to combine certain biosafety program elements
(e.g., Biosafety Manual, [bheh`b\ZelZ_^mrh_\^k (BSO), biosecurity plan) for multiple
containment zones, as determined by an overarching risk assessment. The majority of
requirements in this chapter are to be based on a local risk assessment (LRA) whether it is
indicated in the text or not. Details on the use and interpretation of the matrices that follow
are provided in Chapter 2. A description of the symbols used appears in Section 2.6.
4.1
Biosafety Program Management
A biosafety program is designed to prevent infections, intoxications, and illnesses among

personnel and to protect the community and the environment from harm by preventing
the release of pathogens or toxins. The level of detail and complexity of the biosafety
program will depend on the nature of the organization (i.e., size, structure, and complexity)
and the activities performed within it. A key to the success of any biosafety program is a
strong commitment and involvement by everyone within the organization, including senior
management, supervisors, and individual personnel. The day-to-day management of the
biosafety program can be determined internally and responsibilities delegated accordingly.
4.1
Administrative Controls
4.1.1
A biosafety program to be in place for

the oversight of safety and containment
practices.
50
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4.1
4.1.2
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A biosafety representative(s)
(i.e., designated biological safety
h_\^k [BSO] in licensed facilities)
with the knowledge appropriate for the
containment levels and pathogens and
toxins handled, to be designated for the
oversight of biosafety and biosecurity
practices including:
s VERIFYING THE ACCURACY AND
completeness of licence applications,
animal pathogen import permit
applications, and transfer applications
for the movement of material imported
under the Health of Animals Act (HAA)
and Health of Animals Regulations
(HAR), as applicable;
s COMMUNICATING WITH THE 0UBLIC (EALTH
Agency of Canada (PHAC) and the
Canadian Food Inspection Agency
(CFIA) on behalf of the licence holder
and animal pathogen import permit
holder, as applicable;
s PROMOTING AND MONITORING COMPLIANCE
with applicable legislation (including
the Human Pathogens and Toxins Act
[HPTA], Human Pathogens and Toxins
Regulations [HPTR], HAA, and HAR),
conditions of licence and applicable
conditions of animal pathogen
import permits; applicable biosafety
and biosecurity standards, and the
Biosafety Manual and standard
operating procedures (SOPs), which
includes, but is not limited to;
s arranging and documenting
appropriate biosafety and
biosecurity training for personnel
pertaining to human and
animal pathogens and toxins,
as applicable;
4.1.2 continued on page 52
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CHAPTER 4 OPERATIONAL PRACTICE REQUIREMENTS
4.1
4.1.2 continued from page 51
4.1.2
4.1.3
s informing the PHAC of all

occurrences of inadvertent
possession of a human pathogen
or toxin not already authorized
by the licence;
s informing the PHAC of every
situation where a shipment of
a security sensitive biological
agent (SSBA) has not been
received within 24 hours of when
it was expected;
s conducting periodic inspections
and biosafety audits and reporting
the ndings to the licence holder
and the animal pathogen import
permit holder, as applicable;
s informing the licence holder and
animal pathogen import permit
holder, as applicable, in writing
of any non-compliance by a
person working with human or
animal pathogens, toxins, or other
regulated infectious material
that is not being corrected by that
person after they have been made
aware of it;
s ASSISTING IN THE DEVELOPMENT AND
maintenance of the Biosafety Manual
and SOPs;
s ASSISTING WITH INTERNAL INVESTIGATIONS
of incidents.
Contact information provided to the PHAC
and the CFIA, as applicable, to be kept
up to date.
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4.1
4.1.4
Program intent to be documented and

kept up to date.
4.1.5
Where non-indigenous animal

pathogens are handled, changes in
program intent and changes in SOPs that
impact biocontainment or biosafety to
be submitted to the CFIA for review prior
to implementation.
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Risk Assessments and Planning
4.1.6
An overarching risk assessment to be

conducted and documented to identify
the hazards and appropriate mitigation
strategies for the proposed activities
involving infectious material or toxins.
4.1.7
A biosecurity risk assessment to be

conducted and documented.
4.1.8
A local risk assessment (LRA) to be

conducted to examine each task involving
infectious material or toxins so that the risks
are identied and safe work practices
developed and documented.
4.1.9
A training needs assessment

to be conducted.
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4.1
Biosafety Program Elements
4.1.10
A Biosafety Manual to be developed,

implemented, kept up to date, made
available to personnel inside and outside
of containment zone, and to contain
institutional biosafety policies, programs,
and plans, based on an overarching
risk assessment and LRAs. The Biosafety
Manual to include:
s THE PROGRAM INTENT
s A BRIEF DESCRIPTION OF THE PHYSICAL
design and operation of the
containment zone and systems;
s A DESCRIPTION OF THE
s biosafety program;
s biosecurity plan;
s medical surveillance program;
s training program;
s emergency response plan
(ERP) and incident reporting
procedures;
s housekeeping program;
s facility and equipment maintenance
program for components of the
containment zone, including
integrity testing of primary
containment devices; and
s 3/0S FOR SAFE WORK PRACTICES SPECIlC
to the containment zone.
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4.1
4.1.11
A biosecurity plan, based on a biosecurity

risk assessment, to be developed,
implemented, evaluated and improved
as necessary, and kept up to date. The
biosecurity plan to include mitigation
strategies for the risks associated with:
s PHYSICAL SECURITY
s PERSONNEL SUITABILITY AND RELIABILITY
s ACCOUNTABILITY FOR PATHOGENS TOXINS
and other regulated infectious material;
s inventory;
s INCIDENT AND EMERGENCY RESPONSE AND
s INFORMATION MANAGEMENT
4.1.12
A medical surveillance program, based

on an overarching risk assessment and
LRAs, to be developed, implemented,
and kept up to date.
4.1.13
A respiratory protection program to be

in place when respirators are in use.
4.1.14
A training program, based on a training

needs assessment, to be implemented,
evaluated and improved as necessary,
and kept up to date.
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4.1
4.1.15
SOPs specic to the nature of the work

being conducted in the containment
zone to be developed and documented,
including:
s personal protective equipment
(PPE) requirements;
s ENTRYEXIT PROCEDURES FOR PERSONNEL
animals, and materials;
s USE OF PRIMARY CONTAINMENT DEVICES
s ANIMAL WORK CONSIDERATIONS
s decontamination and waste
management;
s THE SAFE AND SECURE MOVEMENT AND
transportation of infectious material
and toxins, and
s ANY PROCEDURE OR TASK INVOLVING
infectious material, toxins, and/or
infected animals, as determined by
an LRA.
4.1.16
An ERP, based on an overarching risk

assessment and LRAs, to be developed,
implemented, and kept up to date.
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4.2
Medical Surveillance Program
The medical surveillance program aims to prevent and detect illnesses related to
exposure of personnel to pathogens or toxins and to provide a response mechanism
through which potential infections and intoxications can be quickly identied and treated
before serious injury, disease, or transmission to the public occurs.
4.2
Medical Surveillance Program
4.2.1
Liaison to be established with the local

hospital/health care facility.
4.2.2
Containment zone personnel to

immediately inform appropriate internal
personnel or authority of any:
s incident that may have resulted in an
exposure of an individual to a human
pathogen or toxin in a facility; or
s DISEASE THAT MAY HAVE BEEN CAUSED
by an exposure to a human pathogen
or toxin in a facility.
4.2.3
Supervisors to contact any containment

zone personnel with unexpected work
absences.
4.2.4
Emergency medical contact card to be

issued to containment zone personnel
handling non-human primates or a
pathogen identied by a local risk
assessment (LRA).
4.2.5
Emergency medical contact card to be

issued to containment zone personnel.
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4.3
Training Program
Training is a core element of biosafety and biosecurity, and is essential to the success
of the biosafety program. It is critical that personnel be knowledgeable about the
hazards associated with the pathogens and toxins present in the work environment and
the practices and tools that can protect them from these hazards. The training program
encompasses both education (i.e., theoretical) and training (i.e., practical).
4.3
Training Program
4.3.1
Personnel to be trained on the relevant

components of the Biosafety Manual
and standard operating procedures
(SOPs), as determined by the training
needs assessment.
4.3.2
Personnel to be trained on the potential

hazards associated with the work involved,
including the signs and symptoms of
disease(s) caused by the infectious
material or toxins in use and the necessary
precautions to prevent exposure to, or
release of, pathogens or toxins.
4.3.3
Personnel to be trained on the relevant

physical design and operation of the
containment zone and containment
systems.
4.3.4
Personnel to be trained on the correct use

and operation of laboratory equipment,
including primary containment devices.
4.3.5
Personnel working with animals to be

trained in restraint and handling techniques.
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4.3
Training Program
4.3.6
Visitors, maintenance and janitorial staff,

contractors, and others who require
temporary access to the containment
zone to be trained and/or accompanied
in accordance with their anticipated
activities in the containment zone.
4.3.7
Personnel to demonstrate knowledge of

and prociency in the SOPs on which they
were trained.
4.3.8
Trainees to be supervised by authorized

personnel when engaging in activities
with infectious material and toxins until they
have fullled the training requirements.
4.3.9
Review of training needs assessment to

be conducted, at minimum, annually.
Additional or refresher training to be
provided as determined by the review
process or when warranted by a change
in the biosafety program.
4.3.10
Refresher training on emergency response

procedures to be provided annually.
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4.4
Personal Protective Equipment
Personal protective equipment (PPE) includes protective equipment and clothing that
are designed to minimize the risk of exposure to pathogens and toxins. PPE serves as
a last line of defence to prevent exposure in the event of failure in the administrative or
engineering controls. The exception to this is in animal cubicles and post mortem rooms
(PM rooms), where PPE is the primary defence for personnel against exposure. Selection of
PPE is determined by a local risk assessment (LRA) and is specic to both the pathogen(s)
and the work activities to be performed.
4.4
4.4.1
Appropriate dedicated PPE specic to

each containment zone, to be donned
in accordance with entry procedures and
to be exclusively worn and stored in the
containment zone.
4.4.2
Face protection to be used where

there is a risk of exposure to splashes
or ying objects.
4.4.3
Personnel working in animal rooms,

animal cubicles, or PM rooms to wear
dedicated protective footwear and/
or additional protective footwear,
as determined by an LRA.
4.4.4
Gloves to be worn when handling

infectious material, toxins, or
infected animals.
4.4.5
Full body coverage dedicated

protective clothing to be worn inside the
containment barrier where human or
zoonotic pathogens are handled.
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4.4
4.4.6
Dedicated protective clothing and

footwear to be worn inside the
containment barrier where strict animal
pathogens are handled.
4.4.7
An additional layer of protective clothing

to be donned in accordance with entry
procedures prior to work with infectious
material, toxins, or animals infected with
zoonotic pathogens.
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Positive-pressure suits that have passed

integrity testing to be worn inside the
4.4.8
4.4.9
[Not required for laboratory work

areas where infectious material is
exclusively handled in a Class III
biological safety cabinet (BSC) line.]
Respirators to be worn where there is a
risk of exposure to infectious aerosols
that can be transmitted through the
inhalation route or to aerosolized toxins,
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4.5
Entry and Exit of Personnel, Animals, and Material
The operational practices for entry/exit are critical to maintaining containment integrity and
ensuring that only trained and authorized individuals can access the containment zone. In
high containment zones, adherence to operational procedures allows inward directional
Zbkhp (IDA) to be maintained and keeps contaminated or potentially contaminated
personal protective equipment (PPE) inside the containment barrier. The following
matrix presents essential elements for standard operating procedures (SOPs) outlining
entry and exit procedures.
4.5
Entry and Exit of Personnel, Animals,

and Material
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Access and Authorization
4.5.1
Containment zone, animal room,

animal cubicle, and post mortem room
(PM room) doors to be kept closed.
4.5.2
Access to containment zone to be

limited to authorized personnel
and authorized visitors.
4.5.3
Authorization to an individual to access a

part of the facility where security
sensitive biological agents (SSBAs) are
present and accessible to be granted only
after confirmation that a valid Human
Pathogens and Toxins Act 4ecurity
$learance (HPTA 4ecurity $learance) has
been issued to that individual.
4.5.4
An individuals authorization to access a

part of a facility where SSBAs are present
and accessible to be removed when the
individual is no longer in need of access
to that part of the facility or when the
individual no longer holds a valid HPTA
4ecurity $learance.
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4.5

and Material
4.5.5
Access to supporting mechanical and

electrical services for the containment
zone to be limited.
4.5.6
Access to supporting mechanical and

electrical services for the containment
zone to be limited and to be kept
locked at all times.
4.5.7
Access to areas housing an ^_n^gm

decontamination system to be limited
and doors to be kept closed and locked
at all times.
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Entry Procedures
4.5.8
Current entry requirements to be posted

at point(s) of entry to the containment
zone, animal rooms, animal cubicles,
and PM room.
4.5.9
Personnel to verify correct reading

of monitoring device(s) that visually
demonstrate IDA, prior to entry into
area where IDA is provided.
4.5.10
Personal clothing to be stored separately

from dedicated PPE.
4.5.11
Personal belongings to be kept separate

from areas where infectious material or
toxins are handled or stored.
4.5.12
Personal belongings not required for work

to be left outside the containment zone or
in change areas outside the containment
barrier.
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4.5
4.5.13

and Material
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Personnel to doff personal clothing and

footwear and don dedicated clothing and
PPE when entering the containment zone.
Exit Procedures
4.5.14
Personnel to doff dedicated PPE, in a

manner that minimizes contamination
of the skin and hair, when exiting the
containment zone.
4.5.15
Personnel to remove gloves and wash

hands when exiting the containment
zone, animal room, animal cubicle,
or PM room.
4.5.16
Personnel to remove gloves and wash

hands at the containment barrier when
exiting the containment zone, animal
room, animal cubicle, or PM room.
4.5.17
Personnel to doff dedicated PPE (or

additional layer of PPE, when worn)
when exiting the containment barrier of:
s ANIMAL CUBICLES0- ROOMS EXCEPT
when exiting to the dirty corridor; or
s THE CONTAINMENT ZONE
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4.5

and Material
4.5.18
Personnel to decontaminate eyeglasses

at the containment barrier prior to exit,
unless protected from contamination by
additional layer of PPE.
4.5.19
Where non-indigenous animal

pathogens are handled, personnel to
remove all clothing and shower out when
exiting the containment barrier of the
containment zone.
4.5.20
Personnel to remove all clothing and

shower out when exiting:
s THE CONTAINMENT BARRIER OF THE ANIMAL
cubicle/PM room, except when
exiting to the dirty corridor; or
s THE CONTAINMENT BARRIER OF THE
containment zone.
4.5.21
Personnel wearing positive-pressure

suits to proceed through a chemical
decontamination shower, remove
dedicated protective clothing, and
remove all clothing and shower out,
when exiting the containment barrier.
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4.6
Work Practices
The use of safe work practices when handling infectious material or toxins helps protect
personnel from exposure to pathogens and toxins, and helps prevent their release. Good
microbiological laboratory practices are the foundation for all safe work practices
involving biological material. In containment zones where infectious material and toxins
are handled or stored, safe work practices include the proper use and maintenance of
biocontainment systems, biosafety equipment (e.g., biological safety cabinets [BSCs],
centrifuges), as well as aspects of general containment zone maintenance (e.g., tidiness,
clutter). Safe work practices documented in standard operating procedures (SOPs) can
be easily understood and implemented by all personnel.
4.6
Work Practices
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General
4.6.1
Contact of the face or mucous membranes

with items contaminated or potentially
contaminated with pathogens or toxins
to be prohibited.
4.6.2
Hair that may become contaminated

when working in the containment zone
to be restrained or covered.
4.6.3
Type of footwear worn to be selected

to prevent injuries and incidents,
in accordance with containment
zone function.
4.6.4
Personnel to remove jewellery before

entering the containment zone.
4.6.5
Oral pipetting of any substance to

be prohibited.
4.6.6
Open wounds, cuts, scratches, and grazes

to be covered with waterproof dressings.
66
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4.6
Work Practices
4.6.7
Trafc ow patterns from areas of lower

contamination (i.e., clean) to areas of
higher contamination (i.e., dirty) to be
established and followed, as determined
by a local risk assessment (LRA).
4.6.8
Dedicated paper/computer work areas

to be utilized for paperwork and report
writing.
4.6.9
Use of needles, syringes, and other sharp

objects to be strictly limited and avoided
when suitable alternatives are available.
4.6.10
Bending, shearing, re-capping, or

removing needles from syringes to
be avoided, and, when necessary,
performed only as specied in SOPs.
4.6.11
Work surfaces to be cleaned and

decontaminated with a disinfectant
effective against the pathogen(s) in use,
or a neutralizing chemical effective
against the toxin(s) in use, at a frequency
to minimize the potential of exposure to
infectious material or toxins.
4.6.12
Personnel trained and able to provide

immediate emergency assistance to be
available outside the containment zone
when work is being conducted.
4.6.13
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O^kb\Zmbhg of inward directional

Zbkhp (IDA) to be performed routinely,
as described in SOPs.
67
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4.6
Work Practices
4.6.14
Verication of the integrity of primary

containment devices to be performed
routinely, as described in SOPs.
4.6.15
BSCs, when present, to be certied upon

initial installation, annually, and after any
repairs, modication, or relocation.
4.6.16
Operation of containment and life safety

systems to be veried daily, as described
in SOPs.
4.6.17
Integrity of positive-pressure suits to be

veried routinely, as described in SOPs.
Handling Infectious Material and Toxins

4.6.18
Good microbiological laboratory

practices to be employed.
4.6.19
Samples of pathogens, toxins, or other

regulated infectious material to be
opened only in containment zones that
meet the containment level requirements
to which that infectious material or toxin
has been assigned.
4.6.20
Containers of pathogens, toxins, or other

regulated infectious material stored outside
the containment zone to be labelled,
leakproof, impact resistant, and kept either
in locked storage equipment or within an
area with limited access.
68
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4.6
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4.6.21
Containers of pathogens, toxins, or other

regulated infectious material stored outside
leakproof, impact resistant, and kept in
locked storage equipment and within an
area with limited access.
4.6.22
Containers of security sensitive

biological agents (SSBAs) stored outside
leakproof, impact resistant, and kept
in locked storage equipment that is xed
in place (i.e., non-movable).
4.6.23
Pathogens, toxins, and other regulated

infectious material to be stored inside the
containment zone.
4.6.24
Work Practices
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A certied BSC to be used for

procedures involving open vessels
of infectious material or toxins that:
s MAY PRODUCE INFECTIOUS aerosols
or aerosolized toxins, when aerosol
generation cannot be contained
through other methods;
s INVOLVE high concentrations
of infectious material or toxins; or
s INVOLVE large volumes of infectious
material or toxins.
[Not required when collecting samples
from or inoculating animals housed in an
animal cubicle.]
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4.6
4.6.25
Work Practices
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All activities involving open vessels

of infectious material or toxins to be
performed in a certied BSC or other
appropriate primary containment device.
[Not required when collecting samples
from or inoculating animals housed in an
animal cubicle.]
4.6.26
Procedures to be followed to prevent

the inadvertent spread of contamination
from items removed from the BSC after
handling infectious material or toxins.
4.6.27
Personnel to wash hands after completing

tasks that involve the handling of infectious
material or toxins and before undertaking
other tasks in the containment zone.
4.6.28
Centrifugation of infectious material where

inhalation is the primary route of infection
to be carried out in sealed safety cups (or
rotors) that are unloaded in a BSC.
4.6.29
Centrifugation of infectious material to

be carried out in sealed safety cups
(or rotors) that are unloaded in a BSC.
4.6.30
Use of on-demand open ames in a

BSC to be strictly limited and avoided
when suitable alternatives are available;
sustained open ames to be prohibited
in a BSC.
70
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4.6
4.6.31
4.6.32
Work Practices
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Procedures, as determined by an LRA, to

be in place to prevent a leak, drop, spill,
or similar event during the movement
of infectious material or toxins within the
containment zone or between containment
zones within a building.
Large scale cultures of infectious
material or toxins to be contained within
a closed system or other primary
containment device.
4.6.33
Collecting samples, adding materials, or

transferring culture uids from one closed
system to another to be performed in a
manner that prevents the release of aerosols
or the contamination of exposed surfaces.
4.6.34
Experimentally infecting cells or

other specimens derived from the
person conducting the experiment
to be prohibited.
Housekeeping and General Maintenance
4.6.35
Containment zone (including oors) to be

kept clean, free from obstructions, and
free from materials that are in excess,
not required, or that cannot be easily
decontaminated.
4.6.36
Routine cleaning, as described in

SOPs, to be carried out by containment
zone personnel or other staff trained
specically for this task.
71
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4.6
Work Practices
4.6.37
An effective rodent and insect control

program to be maintained.
4.6.38
Water seals in drainage traps to be

maintained through regular usage or lling.
4.6.39
An acceptable mechanism to be utilized

for the safe removal of abà^_\b^g\r
iZkmb\neZm^Zbk!A>I:"em^kl.
4.6.40
A basic tool kit to be available inside

72
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4.7
Animal Work Considerations
Due to their unpredictable behaviour, especially when ill, in vivo work with pathogens
and toxins involving live animals increases the risk associated with any given procedure.
In addition, large volumes of contaminated waste are generated in animal containment
zones. Special considerations and handling techniques for work with animals help prevent
personnel exposure or release of pathogens or toxins where animals are infected or
potentially infected with zoonotic pathogens, or are asymptomatic carriers of human
pathogens. Safe work practices for animal work considerations documented in standard
operating procedures (SOPs) can be easily understood and implemented by all personnel.
4.7
4.7.1
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Proper methods of restraint to be used to

minimize scratches, bites, kicks, crushing
injuries, and accidental self-inoculation.
production areas.]
4.7.2
4.7.3
Primary containment caging housing

infected animals to be identied with labels.
production areas.]
Handling procedures to be employed to
minimize the creation of aerosols and
dissemination of dust from cages, refuse,
and animals.
production areas.]
4.7.4
Entry/exit procedures to be employed

to minimize the release of aerosolized
or airborne pathogens from animal
cubicles and post mortem rooms
(PM rooms).
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4.7
4.7.5
Animals and carcasses to be securely

moved into, out of, and within the
containment zone.
production areas.]
4.7.6
4.7.7
Animal carcasses to be removed from

cubicles/PM rooms via the dirty corridor,
or if removed via the clean corridor, to be
divided into smaller portions (as necessary)
and placed into labelled, leakproof, and
impact resistant transport containers.
Inoculation, surgical, and necropsy
procedures to be designed and carried
out to prevent injuries to personnel and
minimize the creation of aerosols.
production areas.]
4.7.8
Inoculation, surgical, and necropsy

procedures with animals in small animal
containment zones (SA zones) to be
carried out in a certied biological
safety cabinet (BSC) or other appropriate
primary containment device.
production areas.]
4.7.9
Animals to be disinfected and/or cleaned

at site of injection or exposure following
inoculation or aerosol challenge with
production areas.]
74
CL2
CL2Ag
CL3
CL3Ag
CL4
4.8
Decontamination and Waste Management
Effective decontamination of waste is critical in all containment zones so that

contaminated material is treated and safely disposed of. The principles of sterilization,
disinfection, and decontamination are essential for reducing the risk of pathogen
and toxin transmission within containment zones, or release to the environment or the
community. Decontamination and waste management procedures documented in standard
operating procedures (SOPs) can be easily understood and implemented by all personnel.
4.8
4.8.1
Gross contamination to be removed

prior to decontamination of surfaces and
equipment, and disposed of accordingly.
4.8.2
Disinfectants effective against the

pathogen(s) in use and neutralizing
chemicals effective against the toxin(s)
in use to be available and used in the
containment zone.
4.8.3
Sharps to be discarded in containers that

are leakproof, puncture-resistant, and
tted with lids, or specially constructed
for the disposal of sharps waste.
4.8.4
Primary containment devices to be

decontaminated prior to maintenance.
4.8.5
All clothing and personal protective

equipment (PPE) to be decontaminated
when a known or suspected exposure
has occurred.
4.8.6
PPE to be decontaminated prior to disposal

or laundering unless laundering facilities
are located within the containment zone
and have been proven to be effective in
decontamination.
CL2
CL2Ag
CL3
CL3Ag
75
CL4
4.8
4.8.7
Contaminated liquids to be
decontaminated prior to release
to sanitary sewers.
4.8.8
Contaminated equipment, materials,

and waste to be:
s DECONTAMINATED AND LABELLED AS
decontaminated prior to cleaning,
disposal, or removal from the
containment zone or prior to removal
from the animal rooms, animal
cubicles, or post mortem rooms
(PM rooms), as described in SOPs; or
s PLACED IN CLOSED LABELLED AND
leakproof containers that have been
surface decontaminated prior to
removal from the containment zone,
animal rooms, animal cubicles, or
PM rooms, as described in SOPs
for the safe and secure movement
or transportation to a designated
decontamination area or storage
outside of the containment zone.
4.8.9
All equipment, materials, and waste to

be decontaminated at the containment
barrier and labelled as decontaminated
prior to removal from the containment
zone and prior to removal from the animal
rooms, animal cubicles, or PM rooms.
4.8.10
Decontamination technologies and

processes to be validated prior to initial
use and when signicant changes to
the processes are implemented or new
pathogens are introduced.
76
CL2
CL2Ag
CL3
CL3Ag
CL4
4.8
4.8.11
Decontamination technologies and

processes to be routinely veried,
as described in SOPs. Frequency of
o^kb\Zmbhg to be determined by a
local risk assessment (LRA).
4.8.12
Doors of pass-through technologies

not to be opened simultaneously.
4.8.13
Contaminated bedding to be:

s REMOVED AT A ventilated cage
changing station or within a certied
biological safety cabinet (BSC) prior
to decontamination; or
s DECONTAMINATED WITHIN CONTAINMENT
cages.
4.8.14
Animal cubicles, PM rooms, and

dirty corridor, when present, to
be decontaminated when grossly
contaminated and at the end of
an experiment.
4.8.15
Procedures for full room decontamination

to be developed, validated, and followed,
CL2
CL2Ag
CL3
CL3Ag
77
CL4
4.9
Emergency Response
In order to promote personnel safety and the containment of pathogens and toxins, plans need
to be in place for situations where biosafety or biosecurity issues may arise as the result of an
emergency. Emergency situations may include incidents or accidents, medical emergencies,
re, chemical or biological spills, power failure, animal escape, failure of primary containment
devices (e.g., biological safety cabinet [BSC]), loss of containment (e.g., heating, ventilation,
and air conditioning [HVAC]), and natural disasters. Protocols for incident reporting and
investigation are an integral component of an emergency response plan (ERP) as incidents
may be indicative of deciencies in biosafety and/or biosecurity systems.
4.9
Emergency Response
4.9.1
The ERP is to describe emergency

procedures applicable to the
containment zone for:
s ACCIDENTSINCIDENTS
s MEDICAL EMERGENCIES
s lRES
s CHEMICALBIOLOGICAL SPILLS SMALLLARGE
inside/outside BSC and centrifuge);
s POWER FAILURE
s ANIMAL ESCAPE IF APPLICABLE
s FAILURE OF PRIMARY CONTAINMENT DEVICES
s puff-back from class II B2 BSCs,
where present;
s LOSS OF CONTAINMENT
s EMERGENCY EGRESS
s NOTIlCATION OF KEY PERSONNEL AND
relevant federal regulatory agency
(or agencies);
s NATURAL DISASTERS AND
s INCIDENT FOLLOW UP AND RECOMMENDATIONS
to mitigate future risks.
4.9.2
ERP to include procedures for any

infectious material or toxins stored
78
CL2
CL2Ag
CL3
CL3Ag
CL4
4.9
Emergency Response
4.9.3
ERP to include procedures for

positive-pressure suit damage, loss
of breathing air, and loss of chemical
decontamination shower.
4.9.4
ERP to include additional exit procedures

to describe emergency scenarios where
showering out of the containment zone
is necessary.
4.9.5
ERP to include additional emergency

egress procedures for life-threatening
emergencies.
4.9.6
A biological spill kit to be available

inside the containment zone.
CL2
CL2Ag
CL3
CL3Ag
Incident Investigation and Reporting
4.9.7
Incidents involving pathogens, toxins,

other regulated infectious material,
infected animals, or involving failure
of containment systems or control
systems to be reported immediately
to the appropriate internal authority.
4.9.8
Incident investigation to be conducted

and documented for any incident
involving pathogens, toxins, other
regulated infectious material, infected
animals, or failure of containment systems
or control systems, in order to determine
the root cause(s).
79
CL4
4.9
Emergency Response
4.9.9
The Public Health Agency of Canada

(PHAC) to be informed without delay
via the submission of an exposure
ghmb\Zmbhgkîhkm following:
s AN EXPOSURE TO A HUMAN PATHOGEN
or toxin; or
s RECOGNITION OF A DISEASE THAT HAS
or may have been caused by an
exposure to a human pathogen
or toxin.
4.9.10
An exposure follow-up report

documenting the completed investigation,
to be submitted to the PHAC within:
s DAYS OF THE SUBMISSION OF AN
exposure notication report involving
a security sensitive biological
agent (SSBA); or
s DAYS OF THE SUBMISSION OF AN
exposure notication report involving
a human pathogen or toxin other
than an SSBA.
80
CL2
CL2Ag
CL3
CL3Ag
CL4
4.10
Records and Documentation
A biosafety program will generate records for most activities. These records provide
evidence that a specic activity was performed, document the results achieved, and can
also be used for the ongoing improvement of the biosafety program.
4.10
4.10.1
Training and refresher training to be

documented; records to be kept on le.
4.10.2
Inventory of pathogens, toxins, and

other regulated infectious material in
long-term storage to be maintained,
including location and risk group(s).
Inventory to include pathogens, toxins,
and other regulated infectious material
stored outside of the containment zone.
4.10.3
Inventory of Risk Group 3 (RG3) and

Risk Group 4 (RG4) pathogens and
security sensitive biological agent
(SSBA) toxins in long-term storage, both
inside and outside the containment zone,
to be maintained and to include:
s SPECIlC IDENTIlCATION OF THE
pathogens, toxins, and other
regulated infectious material; and
s A MEANS TO ALLOW FOR THE DETECTION
of a missing or stolen sample in a
timely manner.
4.10.4
Drawings and physical specications

(including as built drawings of all
structures and services pertaining to
the containment zone) and reports of
performance and o^kb\Zmbhg tests of
containment systems to be kept on le.
CL2
CL2Ag
CL3
CL3Ag
[Not required for $-SA zones.]
81
CL4
4.10
4.10.5
Records of regular inspections of the

containment zone and corrective actions
to be kept on le.
4.10.6
Records of building and equipment

maintenance, repair, inspection, testing,
or certication, including performance
verication and testing records, in
accordance with containment zone
function, to be kept on le.
4.10.7
Equipment used for performance

verication and testing of containment
systems and essential biosafety
equipment to have a valid calibration
certicate at the time of testing;
calibration certicates to be kept on le.
4.10.8
A record of all individuals entering

and exiting the containment zone to
be maintained and kept on le.
4.10.9
Records of validation and routine

verication of decontamination
technologies and processes to be
kept on le.
82
CL2
CL2Ag
CL3
CL3Ag
CL4
4.10
4.10.10
Records and documentation pertaining to:

s licence activities involving human
pathogens and toxins to be kept on
le for a minimum of 5 years; and
s animal pathogen import permit
requirements for animal pathogens,
toxins, and other regulated infectious
material to be kept on le for a
minimum of 2 years following the date
of disposal, complete transfer, or
inactivation of the imported material.
4.10.11
Records of incidents involving

pathogens, toxins, other regulated
infectious material, infected animals, or
losses of containment to be kept on le
for a minimum of 10 years.
4.10.12
Access to records and documentation

pertaining to controlled activities with
human pathogens and toxins to be
restricted to authorized personnel.
CL2
CL2Ag
CL3
CL3Ag
83
CL4
Chapter 5
PERFORMANCE AND VERIFICATION

TESTINGREQUIREMENTS
CHAPTER 5 PERFORMANCE AND

VERIFICATION TESTING REQUIREMENTS
The requirements presented in Matrices 5.1 through 5.3 describe the minimum performance
and o^kb\Zmbhg tests necessary to demonstrate compliance with the physical containment
requirements outlined in Chapter 3 and, in some cases, with the operational practice
requirements outlined in Chapter 4. Reports demonstrating the successful completion of
these tests are requested by the Public Health Agency of Canada (PHAC) and the Canadian
Food Inspection Agency (CFIA) in support of applications for a licence for controlled
activities with human pathogens and toxins, applications for an animal pathogen
import permit, or _Z\bebmr \^kmb\Zmbhg (or recertication) of containment zones. In
addition, test reports will be monitored by the PHAC and the CFIA for ongoing compliance
verication, including during on-site inspections and audits. The corresponding physical
containment and operational practice requirements from Chapters 3 and 4 are referenced
in the individual explanatory note (found in the Appendix) for each requirement in Matrices
5.1-5.3. The containment level columns indicate the containment levels to which each
requirement applies. In certain scenarios, the PHAC or the CFIA may request additional
tests, on a case-by-case basis, to demonstrate performance or verication of containment
systems other than those tests described in the matrices below. A description of the symbols
used appears in Section 2.6.
5.1
Performance and Verication Tests for

All Containment Levels
The following matrix describes the minimal performance and o^kb\Zmbhg tests to be performed
by all types of work areas at all containment levels (CL2-CL4).
5.1
5.1.1
I^k_hkfZg\^Zg]O^kb\ZmbhgM^lml_hk
Performance and verication tests

described in 5.1.2-5.1.7 to be conducted
and documented at minimum annually, or
more frequently as indicated by:
s A CHANGE REPAIR OR MODIlCATION
to the containment system;
s A CONDITION OF licence; or
s A REQUEST OF THE 0UBLIC (EALTH !GENCY
of Canada (PHAC) or the Canadian
Food Inspection Agency (CFIA).
86
CL2
CL2Ag
CL3
CL3Ag
CL4
5.1
5.1.2
Visual inspections of the containment

zone to be conducted in order to identify
faults and/or deterioration; when found,
corrective actions to be taken.
5.1.3
Visual inspections of small in-line lters to

be conducted and lters to be replaced in
accordance with maintenance schedules
or as necessary to maintain function.
5.1.4
Decontamination technologJFT and

processes to be validated through the use
of representative loads in conjunction
with application-specic biological
indicators, chemical integrators, and/or
parametric monitoring devices consistent
with the technology/method.
5.1.5
Class II biological safety cabinets (BSCs)

to be certied in accordance with NSF/
ANSI 49, where possible.
CL2
CL2Ag
CL3
CL3Ag
87
CL4

5.1
5.1.6
Verication of the following manufacturers

specications to be demonstrated, where
the design of a BSC or custom ventilated
enclosure does not permit certication in
accordance with NSF/ANSI 49:
s INTEGRITY OF THE abà^_\b^g\r
iZkmb\neZm^Zbk!A>I:"em^kl to be
tested in accordance with the HEPA
lter test method IEST-RP-CC034.3
or equivalent;
s MAINTENANCE OF A MINIMUM AVERAGE
inow velocity of 0.38 m/s (75 ft/
min) through the front opening during
normal operation to be veried;
s AIRmOW PATTERN INSIDE THE CABINET AND
at access opening to ensure no back
streaming of air to be demonstrated;
s INTEGRITY OF "3#S DESIGNED WITH
positive-pressure plenums to be
demonstrated by determining that
exterior surfaces of all plenums,
welds, gaskets, and plenum
penetrations or seals, are free
of leaks (to be performed during
initial installation, if any panels
are removed, or if the cabinet is
relocated); and
s ALARMS TO BE DEMONSTRATED TO FUNCTION
as intended.
5.1.7
Integrity of primary containment

devices other than BSCs (e.g., process
equipment, closed systems, primary
containment caging) to be tested in
accordance with testing procedures and
acceptance criteria appropriate for the
equipment and design.
88
CL2
CL2Ag
CL3
CL3Ag
CL4
5.2
Additional Performance and Verication Tests

for Select CL2 and CL2- Ag Zones and All
CL3-CL4 Zones
In addition to the performance and o^kb\Zmbhg tests to be performed for all containment
zones outlined in Matrix 5.1, the following matrix describes the additional performance
and verication tests to be performed by any containment zone at all containment levels
CL2-CL4, with the exception of CL2 laboratory work areas and CL2 SA zones, unless,
as indicated, prions or security sensitive biological agents (SSBAs) are handled in these
work areas.
5.2
5.2.1
:]]bmbhgZeI^k_hkfZg\^Zg]O^kb\Zmbhg
Tests for Select CL2 and CL2-Ag Zones
and All CL3-CL4 Zones

described in 5.2.3-5.2.11 to be
conducted and documented at minimum
every two years, or more frequently,
where specied or as indicated by:
s A CONDITION OF licence;
s A CONDITION OF animal pathogen
import permit; or
s A REQUEST OF THE 0UBLIC (EALTH !GENCY
of Canada (PHAC) or the Canadian
Food Inspection Agency (CFIA).
CL2
CL2Ag
CL3
CL3Ag
[Does not apply where SSBAs are present.]
5.2.2

described in 5.2.3-5.2.15 to be
conducted and documented at
minimum annually, or more frequently
as indicated by:
s A CONDITION OF LICENCE OR
s A REQUEST OF THE 0(!# OR THE #&)!
89
CL4

5.2
5.2.3
Operation of controlled access systems

and security systems to be veried to be
functioning as intended.
CL2
CL2Ag
PS
[Not required in CL2 SA zones where

Prions and SSBAs are not present.]
5.2.4
Emergency power and uninterruptJCMe

power supply (UPS) systems to be tested
under representative electrical load
conditions.
PS

Prions and SSBAs are not present.]
5.2.5
5.2.6
Operation of communication systems

to be veried.
Prions are not present.]
Integrity of penetrations of the
containment barrier, seals, and
surfaces, to be visually inspected.
Prions are not present.]
5.2.7
Testing to be performed at all critical

doors on the containment barrier, where
bgpZk]]bk^\mbhgZeZbkhp (IDA) is
provided, to verify, using a smoke pencil or
other visual aid that does not inuence the
direction of airow, that IDA is maintained
in accordance with facility design.
[Not required in CL2 SA zones.]
90
CL3
CL3Ag
CL4
5.2
CL2
CL2Ag
5.2.8
Abà^_\b^g\riZkmb\neZm^Zbk!A>I:"
em^kl to be tested in situ by particle
challenge testing using the scanning
method in accordance with IEST-RPCC034.3 or IEST-RP-CC006.3. When
scan testing is not possible, probe testing
is acceptable.
5.2.9
Operation of mechanical or electronic

door interlocks and associated manual
overrides to be veried.
5.2.10
Decontamination technology alarms

to be veried to function as intended.
5.2.11
Water supply backow preventers to be

tested at minimum annually in accordance
with CAN/CSA B64.10/B64.10.1.
5.2.12
Integrity of the seals of containment

barrier penetrations, animal cubicle
penetrations, and post mortem room
(PM room) penetrations to be tested with
a smoke pencil or other aid that does not
inuence the direction of airow.
CL3
CL3Ag
Compressed breathing air and

systems to be veried in accordance
with CAN/CSA-Z180.1.
5.2.13
[Not required for CL4 laboratory work

areas where infectious material
is exclusively handled in a Class III
biological safety cabinet (BSC) line.]
91
CL4

5.2
5.2.14
5.2.15
Positive-pressure suits to be tested to

ensure operation in accordance with
manufacturers specications.
Chemical decontamination shower
systems, including the disinfectant tank
low level alarm and parametric monitoring
and recording devices, to be veried to
function as intended.
92
CL2
CL2Ag
CL3
CL3Ag
CL4
5.3
Performance and Verication Tests to be Conducted

on Commissioning of Containment Zone
In addition to the performance and o^kb\Zmbhg tests to be performed outlined in Matrices

5.1 and 5.2, the following matrix describes the performance and verication tests
to be performed by any containment zone at the indicated containment levels upon
commissioning of the containment zone. Test reports documenting successful completion
of these tests will be requested by the Public Health Agency of Canada (PHAC) and the
Canadian Food Inspection Agency (CFIA) for a newly commissioned containment zone to
support an application for a new licence for controlled activities with human pathogens
and toxins, for an application for an animal pathogen import permit for a containment
zone that has not previously imported an animal pathogen, or for the initial facility
\^kmb\Zmbhg for a containment zone intending to conduct activities with non-indigenous
animal pathogens. Retesting of the containment system(s) is indicated by a change,
repair, or modication to the containment device or system; a condition of licence; or a
request of the PHAC or the CFIA.
5.3
I^k_hkfZg\^Zg]O^kb\ZmbhgM^lml
to be Conducted on Commissioning
of Containment Zone
5.3.1
Drain piping leading to an ^_n^gm

decontamination system to be tested in
accordance with the National Plumbing
Code of Canada. Air test on drainage
system to be performed at standard code
pressure specications of 35,000 Pa (i.e.,
141 inches of water gauge [in. w.g.]).
5.3.2
Integrity of abà^__b\b^g\riZkmb\neZm^
BJS )&1"
GJMUFS housings to be tested
in situ by pressure decay in accordance
with ASME N511; test pressure to be
determined in accordance with ASME
AG-1. Acceptance criteria include the
provision that the rate of leakage is not
to exceed 0.1% of volume/minute at
1,000 Pa (i.e., 4 inches water gauge
[in. w.g.]) minimum test pressure.
CL2
CL2Ag
CL3
CL3Ag
93
CL4

5.3
I^k_hkfZg\^Zg]O^kb\ZmbhgM^lml
to be Conducted on Commissioning
of Containment Zone
5.3.3
Heating, ventilation, and air conditioning

(HVAC) system and controls to be
veried during scenarios simulating
failure of system components, including
exhaust fan(s), supply fan(s), power, and
Class II B2 biological safety cabinet
(BSC) exhaust fan(s) (where present), as
determined by containment zone design.
Acceptance criteria include demonstration
that reversal of inward directional
Zbkhp (IDA) is not sustained at critical
doors; Class II B2 BSC puff-back is
minimized, and HVAC system alarms
and interlocks operate as intended.
5.3.4
Supply ductwork, located between

backdraft protection and containment
barrier, and exhaust ductwork, located
between containment barrier and HEPA
lter or isolation damper, to be visually
inspected and tested in situ by pressure
decay in accordance with ASME
N511; test pressure to be determined
in accordance with ASME AG-1.
Acceptance criteria include the provision
that the rate of leakage is not to exceed
0.1% of volume/minute at 1,000 Pa
(i.e., 4 in. w.g.) minimum test pressure.
5.3.5
Integrity of containment barrier to be tested

by pressure decay testing. Acceptance
criteria include two consecutive tests with
a maximum of 250 Pa (i.e., 1 in. w.g.)
loss of pressure from an initial 500 Pa
(i.e., 2 in. w.g.) over a 20 minute period.
[Not required in CL3-Ag zones where
only human and/or indigenous animal
pathogens are handled.]
94
CL2
CL2Ag
CL3
CL3Ag
CL4
REFERENCES
REFERENCES
Legislation
Health of Animals Act (S.C. 1990, c. 21). (20).
Health of Animals Regulations (C.R.C., c. 296). (201).
Human Pathogens and Toxins Act (S.C. 2009, c. 24). (20).
Human Pathogens and Toxins Regulations (SOR/2015-44). (2015).
Human Pathogens Importation Regulations (SOR/94-558). (1994) (repealed 2015).
Guidance Documents
"VTUSBMJB(SPVQ
-JTUPG)VNBOBOE"OJNBM1BUIPHFOTBOE5PYJOTGPS&YQPSU
$POUSPM. 3FUSJFWFE GSPNIUUQXXXBVTUSBMJBHSPVQOFU
FOIVNBO@BOJNBM@QBUIPHFOTIUNM.
Canadian Commission on Building and Fire Codes, & the National Research Council
Canada. (2010). National Plumbing Code of Canada, 2010 (9th ed.).
Ottawa, ON, Canada: Institute for Research in Construction, National
Research Council Canada.
Canadian Food Inspection Agency. (2010). Containment Standards for Facilities
Handling Aquatic Animal Pathogens (1st ed.). Ottawa, ON, Canada:
Canadian Food Inspection Agency.
Canadian Food Inspection Agency. (2007). Containment Standards for Facilities
Handling Plants Pests (1st ed.). Ottawa, ON, Canada: Canadian Food
Inspection Agency.
Canadian Food Inspection Agency. (2005). Containment Standards for Laboratories,
Animal Facilities and Post Mortem Rooms Handling Prion Disease Agents.
Ottawa, ON, Canada: Canadian Food Inspection Agency.
Canadian Food Inspection Agency. (1996). Containment Standards
for Veterinary Facilities (1st ed.). Ottawa, ON, Canada:
Canadian Food Inspection Agency.
Government of Canada. (2015). Canadian Biosafety Handbook (2nd ed.).
Ottawa, ON, Government of Canada.
Government of Canada. (2013). Canadian Biosafety Standards and Guidelines
(1st ed.). Ottawa, ON, Government of Canada.
Public Health Agency of Canada. (2004). Laboratory Biosafety Guidelines
(3rd ed.). Ottawa, ON, Canada: Public Health Agency of Canada.
8PSME0SHBOJTBUJPOGPS"OJNBM)FBMUI
0*&-JTUFEEJTFBTFT JOGFDUJPOTBOE
JOGFTUBUJPOT3FUSJFWFE GSPNIUUQXXXPJFJOUBOJNBM
IFBMUIJOUIFXPSMEPJFMJTUFEEJTFBTFT
96
Technical Standards and Codes

ANSI/SMACNA 016-2012, HVAC Air Duct Leakage Test Manual, 2nd Edition.
(2012). Chantilly, VA, USA: Sheet Metal and Air Conditioning Contractors
National Association, Inc.
ASME AG-1-2012, Code on Nuclear Air and Gas Treatment. (2012).
New York, NY, USA: American Society of Mechanical Engineers.
ASME N511-2007, In-service Testing of Nuclear Air Treatment, Heating,
Ventilating, and Air-Conditioning Systems. (2007). New York, NY, USA:
American Society of Mechanical Engineers.
CAN/CSA B64.10-11/B64.10.1-11, Selection and Installation of Backow
Preventers/Maintenance and Field Testing of Backow Preventers. (2011).
Mississauga, ON, Canada: Canadian Standards Association.
CAN/CSA Z180.1-13 (R2013), Compressed Breathing Air and Systems.
(2000). Mississauga, ON, Canada: Canadian Standards Association.
IESTRP-CC001.5, HEPA and ULPA Filters. (2010). Rolling Meadows, IL, USA:
Institute of Environmental Sciences and Technology.
IESTRP-CC0,5FTUJOH$MFBOSPPNT. (200). Rolling Meadows, *- 64"
Institute of Environmental Sciences and Technology.
IESTRP-CC034, HEPA and ULPA Filter Leak Tests. (2009). Rolling Meadows,
IL, USA: Institute of Environmental Sciences and Technology.
NSF/ANSI 49-201, Biosafety Cabinetry: Design, Construction, Performance,
and Field Certification. (201). Ann Arbor, MI, USA: National Sanitation
Foundation / American National Standards Institute.
97
APPENDIX
APPENDIX EXPLANATORY NOTES

The following table provides additional information pertaining to the physical containment,
operational practice, and performance and verication testing requirements outlined in
Chapters 3, 4, and 5, respectively. Specically, these notes include a brief explanation
of the risk(s) mitigated by a requirement and typical examples of how the requirement may
be achieved. Further guidance on the subjects can be found in the Canadian Biosafety
Handbook, 2nd Edition.
Requirement
Number(s)
Explanatory Note
3.1.1
A door is a physical barrier that protects against the release of infectious

material or toxins by separating the containment zone (i.e., the dirty or
contaminated area) from public and administrative areas (i.e., clean
or uncontaminated areas) while also providing a security barrier to limit
access to the zone.
3.1.2
Separating dedicated containment zone paper/computer work stations

from benches and other areas where infectious material and toxins are
actively handled minimizes the risk of contamination of ofce materials
which may be difcult to decontaminate (e.g., paper, notebooks) or
may become damaged by decontamination (e.g., electronic devices).
This can be achieved by locating these stations in a dedicated room
within the containment zone, by installing a physical partition (e.g.,
splash shield) between a paperwork station adjacent to a laboratory
bench, or by locating paper/computer work stations in a space inside
the containment zone but at a distance from benches.
3.1.3
The design and choice of materials used for walls, ceilings, oors,
and barrier devices are critical for ensuring that the containment zone
has the structural stability to withstand both internal stresses, such
as the negative or positive pressures that result from a failure of the
supply air or exhaust air fan, and external stresses, such as external
weather conditions (e.g., temperature extremes, humidity, wind),
environmental disasters (e.g., tornado, earthquake), and security
threats (e.g., break-in, theft). This is best achieved at the design stage
by locating the containment zone away from external envelope walls
and choosing construction materials in accordance with anticipated
function of the zone.
100
Requirement
Number(s)
Explanatory Note
3.1.4
Separating laboratory work areas inside LA zones from spaces where

animals are housed (i.e., animal cubicles) prevents contamination of
work materials not associated with animal work. This can be achieved
by designing the animal containment zone to include a laboratory work
area for activities that do not directly involve the animals (e.g., preparing
samples or inoculants).
3.1.5
Cold storage equipment located in, or adjacent to, the PM room is

important to protect against the putrefaction of animal carcasses during
temporary storage while awaiting necropsy or disposal. This can be
achieved by including an integral cold room (suitable for large-sized
animals) or installing a refrigerator or freezer of adequate size.
3.2.1
Basic pest control and security measures on windows that can open
protect against the entry of small-sized animals and insects and prevent
the release of infectious material out of the containment zone, especially
where the window opens directly to the outdoors. This can be achieved
by tting windows with properly installed screens that are in good repair
and by closing and locking windows to prevent unauthorized entry, in
particular, when the containment zone is unoccupied.
3.2.2
Keeping windows on the containment barrier that can open locked in a

closed position protects against unauthorized entry to the containment zone.
3.2.3
Sealed windows protect against unauthorized entry and maintain the air
pressure differentials in containment zones where IDA is provided.
3.2.4
Glazing materials, such as double-glazed windows, tempered glass,

wire glass, heat or chemically strengthened glass, laminated safety
glass, acrylic glazing, polycarbonate glazing, and adhesive window
security lms, provide various degrees of protection against breakage,
security threats, or environmental threats.
3.2.5
Windows located on the containment barrier that allow the public to

see into animal rooms, animal cubicles, or PM rooms pose a biosecurity
risk and compromise animal well-being.
101
Requirement
Number(s)
Explanatory Note
3.2.6
Ensuring that equipment on the containment barrier maintains a

continuous seal with the adjacent wall, ceiling, or oor protects the
integrity of the containment barrier and protects against the inadvertent
release of infectious material or toxins. A bioseal (i.e., a biological
sealing ange or gasket made of exible material that surrounds the
body of the barrier autoclave or other equipment on the containment
barrier and creates a hermetic seal between the clean and dirty sides
of the containment zone) is an example of a treatment that can be used
to create a seal around a barrier autoclave or other equipment that
penetrates the containment barrier.
3.2.7
Preventing the simultaneous opening of doors on both sides of passthrough technologies prevents a containment breach and the release
of infectious material or toxins from the containment zone. Examples
of suitable mechanisms at CL3 include mechanical or electronic door
interlocks, visual or audible alarms, or SOPs preventing personnel from
opening doors simultaneously.
3.2.8
Preventing the simultaneous opening of doors on both sides of pass-through

technologies prevents a containment breach and the release of infectious
material or toxins from the containment zone. Examples of suitable
mechanisms at CL3-Ag include mechanical or electronic door interlocks.
3.2.9
Preventing the simultaneous opening of doors on both sides of passthrough technologies prevents a containment breach and the release
of infectious material or toxins from the containment zone. A backup
mechanism is included at CL4 to provide added protection. Examples
of suitable mechanisms at CL4 include mechanical or electronic door
interlocks, and visual or audible alarms to prevent personnel from
opening doors simultaneously.
102
Requirement
Number(s)
Explanatory Note
3.2.10
Penetrations of the containment barrier at or below the work surface

and any other surface that may become contaminated (e.g., conduits,
plumbing, and wiring) may leave gaps between the penetration and
the surface of the containment barrier that could allow the inadvertent
release of infectious material. Sealing these gaps with sealants that
are non-shrinking and compatible with the chemical disinfectants in use
(e.g., silicone, polyurethane, or polyether caulking) allows for proper
spill cleanup and surface decontamination, and maintains the integrity
of the containment barrier.
3.2.11
Penetrations of the containment barrier (e.g., dunk tanks, pass-through

chambers, conduits, plumbing, and wiring) may leave gaps between
the penetration and the surface of the containment barrier that could
allow the inadvertent release of infectious material. Sealing these gaps
with sealants that are non-shrinking and compatible with the chemical
disinfectants in use (e.g., silicone, polyurethane, or polyether caulking)
allows for proper spill cleanup and surface decontamination, and
maintains the integrity of the containment barrier.
3.2.12
Penetrations of the containment barrier of animal cubicles and PM room

(e.g., dunk tanks, pass-through chambers, conduits, plumbing, and wiring)
may leave gaps between the penetration and the surface of the containment
barrier that could allow the inadvertent release of infectious material.
Sealing these gaps with sealants that are non-shrinking and compatible with
the chemical disinfectants in use (e.g., silicone, polyurethane, or polyether
caulking) allows for proper spill cleanup and surface decontamination, and
maintains the integrity of the containment barrier.
3.3.1
Lockable doors provide a basic security barrier to prevent unauthorized

access to the containment zone and to safeguard the infectious material
and toxins stored inside.
103
Requirement
Number(s)
Explanatory Note
3.3.2
Biohazard warning signage is a critical messaging tool designed to

advise that infectious material or toxins are present in the containment
zone, to indicate any special entry requirements, and to provide contact
information in case of an emergency.
3.3.3
Project-specic signage posted at the point(s) of entry for animal rooms,

animal cubicles, and PM rooms is an important messaging tool to inform
of any special entry requirements for a particular experiment or study
where hazards unique to a room or cubicle exist (i.e., the same hazard
is not present in the other adjoining rooms within the containment zone).
3.3.4
Controlled access systems are used to provide restricted access to the

part of the containment zone where SSBA are present and accessible to
authorized individuals only, and to maintain the security of that part of the
containment zone at all times (i.e., locked at all times) when SSBA are
present and accessible (including when vacant and during emergency
evacuation situations). Examples of controlled access systems include
biometric readers, electronic access card systems, keypads, key code
systems, key-locks with non-reproducible keys, or an equivalent system.
3.3.5
Controlled access systems are used to provide restricted access to the

containment zone to authorized individuals and to maintain the security
of the containment zone at all times (including when vacant and during
emergency evacuation situations). Examples of controlled access
systems include biometric readers, electronic access card systems,
keypads, key code systems, key-locks with non-reproducible keys, or an
equivalent system.
3.3.6
Controlled access systems are used to provide restricted access to

animal cubicles and PM rooms to authorized individuals and to maintain
the security of those areas at all times (including when vacant and
during emergency evacuation situations). Examples of controlled access
systems include biometric readers, electronic access card systems,
keypads, key code systems, key-locks with non-reproducible keys, or an
equivalent system. Physical systems are preferred over other mechanisms
to restrict access.
104
Requirement
Number(s)
Explanatory Note
3.3.7
Keys that cannot be reproduced without the authorization of the

containment zone director, supervisor, manager, or other delegated
individual (as determined by the organization) protects against
unauthorized entry and restricts access to the containment zone to only
authorized personnel to whom keys have been issued.
3.3.8
Providing backup to a controlled access system maintains the security

of the containment zone, even in the event of a power failure or
emergency release of electronic locking systems. Examples of alternate
controlled access systems include a physical key code system or keylocks with non-reproducible keys.
3.3.9
Dedicated storage space is necessary to store the PPE used in the

containment zone (e.g., lab coat, coveralls, face shields, respirators)
and to separate these items from personal clothing (e.g., coats, hats,
boots) in order to prevent contamination. Hooks, lockers, shelves, or
spaces within dedicated change areas are examples of dedicated
storage space for PPE.
3.3.10
Dedicated clothing change areas at the entry and exit to the containment
zone provide the space necessary to don and doff dedicated PPE,
including protective clothing, used inside the containment zone and
protects against the contamination of personal clothing. The clothing
change areas may be an anteroom, part of an anteroom, or, in some
cases, a designated area at the entry and exit of the containment zone
that is separated into a clean change area and dirty change area
by a line demarcating the clean and dirty areas.
3.3.11
The presence of an anteroom (or anterooms) at the point(s) of personnel

entry to and exit from a containment zone creates a buffer space to
maintain IDA created through the negative air pressure differentials and
prevents the migration of potentially contaminated air to an area of
lower contamination or lower containment (i.e., outside the containment
barrier or outside the containment zone). This may be achieved by a
single anteroom for entry and exit for the containment zone or multiple
anterooms dedicated for the entry and exit of personnel separate
from the entry and exit of animals or equipment, as determined by the
function and activities of the containment zone.
105
Requirement
Number(s)
Explanatory Note
3.3.12

entry to and exit from a containment zone or each animal cubicle
and PM room creates a buffer space to maintain IDA created through
the negative air pressure differentials and prevent the migration of
potentially contaminated air to an area of lower contamination or
lower containment (i.e., outside the containment barrier of the animal
cubicle or PM room, or outside the containment zone). This may be
achieved by a single anteroom for entry and exit for the containment
zone or multiple anterooms dedicated for the entry and exit of personnel
separate from the entry and exit of animals or equipment, as determined
by the function and activities of the containment zone.
3.3.13

entry to and exit from a containment zone and each animal cubicle
and PM room creates a buffer space to maintain IDA created through
the negative air pressure differentials and prevents the migration of
potentially contaminated air to an area of lower contamination or
lower containment (i.e., outside the containment barrier of the animal
cubicle or PM room, or outside the containment zone). This may be
achieved by a single anteroom for entry and exit for the containment
zone or multiple anterooms dedicated for the entry and exit of personnel
separate from the entry and exit of animals or equipment, as determined
by the function and activities of the containment zone.
3.3.14
A walk-through body shower, located on the containment barrier of the

containment zone between the clean and dirty change areas in the
anterooms, allows personnel to wash their hair and body to remove any
potential contamination before exiting the containment zone. The body
shower is considered to be an extension of the dirty change area,
and is inside the containment barrier.
3.3.15
A walk-through body shower, located on the containment barrier of the

containment zone and another located on the containment barrier of
each animal cubicle and PM room, between the clean and dirty
change areas in the respective anterooms, allows personnel to wash
their hair and body to remove any potential contamination before
exiting the containment barrier of the animal cubicle, PM room, or
containment zone. The body shower is considered to be an extension of
the dirty change area, and is inside the containment barrier.
106
Requirement
Number(s)
Explanatory Note
3.3.16
In CL4 zones where personnel wear positive-pressure suits, a chemical

shower (or suit decontamination shower) is a critical safety feature to be
included in the dirty change area of the anteroom to decontaminate
the suit before it is removed. The location of the chemical shower in
the exit sequence is critical to prevent exposure of personnel. The
chemical shower is located at the immediate exit from the contaminated
area, followed by the designated suit change area(s) (i.e., dirty
change areas where the decontaminated suit is removed), then by
the walk-through body shower(s) on the containment barrier which
allows personnel to wash their hair and body to remove any potential
contamination, and, nally, by clean change area(s) just before the
exit from the containment zone. The chemical decontamination shower,
suit change area, and walk-through body shower are considered to be
an extension of the dirty area within the anteroom, and are inside the
3.3.17
Preventing the simultaneous opening of anteroom critical door(s) with

adjacent and sequential anteroom doors maintains the negative air
pressure differentials and limits the migration of potentially contaminated
air into the clean change area or outside the containment zone.
Examples of suitable mechanisms include physical means, such as
mechanical or electronic door interlocks (equipped with manual
overrides for emergency exit), visual or audible alarms, or, operational
means, such as SOPs and signs preventing personnel from opening
doors simultaneously.
3.3.18
Mechanical or electronic door interlocks (equipped with manual

overrides for emergency exit) prevent the simultaneous opening of
anteroom critical door(s) with adjacent and sequential anteroom doors,
which maintains the negative air pressure differentials and limits the
migration of potentially contaminated air into the clean change area
or outside the containment zone.
3.3.19
Preventing the simultaneous opening of chemical decontamination

shower doors maintains the negative air pressure differentials and limits
the migration of potentially contaminated air into the change areas of
the anterooms.
107
Requirement
Number(s)
Explanatory Note
3.3.20
Sealable doors (e.g., three-sided or four-sided gasket, four-sided door

jamb) are designed to allow leakage of air under normal operating
conditions yet are capable of being sealed to isolate individual animal
cubicles and PM rooms for gaseous decontamination and pressure
decay testing when needed.
3.3.21
Airtight doors (e.g., inatable air pressure resistant or compression

seals) are designed to ensure 0% leakage of air under normal operating
conditions to maintain the integrity of the containment barrier and to
withstand pressure decay testing and gaseous decontamination.
3.4.1
Cleanable and resistant surface materials and nishes (e.g., paint,

epoxy, and other protective nishes) provide protection against the
stresses associated with activities performed inside the containment
zone, which may include repeated decontamination (e.g., chemical,
gaseous), frequent high pressure washing in animal containment zones,
and activities causing impacts and scratches (e.g., movement of largesized animals across oors, equipment resting on surfaces, animal
cages). Other examples of surfaces that may become contaminated as
a result of the procedures in use or spills of infectious material or toxins,
and require decontamination, include animal holding units, shelves, and
interiors of drawers and cabinets, as determined by the activities and
function of the containment zone. Non-absorbent materials may include
stainless steel, epoxy resin surfaces or chemical resistant plastic laminate
for benchtops, and urethane or vinyl for stools and chairs.
3.4.2
The continuity of adjacent surfaces (e.g., walls and oors, benchtops

and other work surfaces) and overlapping material (e.g., ooring,
baseboards, coving, backsplashes) provides a continuous barrier
designed to prevent contaminated liquids from reaching surfaces that
are hard to access and decontaminate. Heat welding or a continuous
bead of silicone sealant, such as caulking, can be used to create a
continuous surface.
3.4.3
Smooth rims and corners reduce the risk of punctures or tears in positivepressure suits that could result in exposure to pathogens or toxins.
108
Requirement
Number(s)
Explanatory Note
3.4.4
Backsplashes that are continuous or sealed at the junction between

the wall and bench provide a continuous barrier designed to prevent
contaminated liquids from reaching surfaces that are hard to access and
decontaminate. Heat welding or a continuous bead of silicone sealant
as caulking can be used to create a continuous surface.
3.4.5
Slip-resistant oors (e.g., textured surfaces) help prevent slips and falls
and the associated risk of exposure to infectious material or toxins (e.g.,
via a splash, spill, inoculation or scratch). Different rooms or spaces may
require a different degree of slip resistance (i.e., coefcient of friction),
as determined by the activities and function of the different spaces
inside the containment zone (e.g., animal room versus a storage room).
3.4.6
Appropriate oor design (including oor joists, spacing, and trusses)

and materials allow ooring to resist damage and withstand the
anticipated loads represented by heavy animals and caging equipment,
as applicable.
3.4.7
The continuity of the seal between the oor and wall allows liquids on
the oor to be contained and facilitates decontamination after a spill in
a laboratory work area and routine cleaning and decontamination of
animal rooms and cubicles. Heat welding or continuous bead of silicone
sealant, such as caulking, can be used to create a continuous surface.
3.4.8
The continuity of the seal between the wall and ceiling facilitates
maintenance, cleaning, and decontamination. Heat welding or continuous
bead of silicone sealant, such as caulking, can be used to create a
continuous surface.
3.4.9
Interior surfaces (i.e., walls, oors, ceiling) made of materials that limit
the penetration of gases and liquids (e.g., stainless steel, epoxy resin
or laminate covering, non-porous materials) provide room integrity,
facilitate surface and room decontamination, and serve to contain any
large volumes of contaminated liquids that may be present (e.g., animal
wastes, large scale process uids).
109
Requirement
Number(s)
Explanatory Note
3.4.10
Minimizing and appropriately shielding protruding obstructions (e.g.,

pipes, conduits) prevents injury to animals and personnel. It also helps
prevent containment breaches caused by damage to the protruding
obstruction by a large-sized or distressed animal.
3.5.1
IDA, established by HVAC system design through negative air pressure

differentials, ensures that air ows from areas of lower containment or
low risk of contamination to areas of higher containment (i.e., higher risk
of contamination), never the reverse. This prevents the release and the
spread of contamination to lower levels of containment by establishing
a physical containment barrier of air against airborne or aerosolized
infectious material or toxins. For example, IDA forces air to ow from
the clean change area into the dirty change area, from the dirty
change area into the laboratory work area, and from the laboratory
work area into the animal cubicle.
3.5.2
IDA, established by HVAC system design through negative air pressure

differentials, ensures that air ows from areas of lower containment or
low risk of contamination to areas of higher containment (i.e., higher risk
of contamination), never the reverse. This prevents the release and the
spread of contamination to lower levels of containment by establishing
a physical containment barrier of air against airborne or aerosolized
infectious material or toxins. For example, IDA forces air to ow from
the clean change area into the dirty change area, from the dirty
change area into the laboratory work area, and from the laboratory
work area into the animal cubicle.
3.5.3
Monitoring devices that visually demonstrate IDA (e.g., EJGGFSFOUJBM

QSFTTVSF gauge, floating ball, alarms) allow personnel to verify that
the HVAC system is working properly and that IDA is being maintained.
3.5.4
Pressure differential monitoring lines that penetrate the containment barrier

designed to prevent the migration of potentially contaminated air (e.g.,
provided with HEPA ltration, small in-line lters, or fully sealed) protect
against the release of aerosolized or airborne infectious material or toxins
in the event of positive pressurization due to HVAC system failure.
110
Requirement
Number(s)
Explanatory Note
3.5.5
Alarms that indicate HVAC system failure (e.g., strobic visual alarms,
intercom audible alarms) are critical to protect and enable personnel,
both inside and outside the containment zone, to quickly initiate
emergency procedures and repairs to prevent containment breaches.
3.5.6
Independent supply and exhaust air systems prevent the contamination

of areas outside the containment zone in the event of HVAC system
failure or airow reversal. Where air systems in CL3 are combined with
those of other areas, HEPA lters or isolation dampers (e.g., gas-tight
dampers, bubble-tight dampers) are examples of backdraft protection
that can be used to prevent contaminated air from reaching areas of
lower containment.
3.5.7
Backdraft prevention on supply air ductwork prevents the contamination

of supply air ductwork outside of the containment barrier in the event
of HVAC system failure or airow reversal. HEPA lters or isolation
dampers (e.g., gas-tight damper, bubble-tight damper) are examples of
backdraft protection that can be used on the supply air duct.
3.5.8
HEPA lters provide backdraft protection and prevent infectious material

and toxins from being released in the event of HVAC system failure or
airow reversal.
3.5.9
Air system interlocks shutdown or divert the supply air system

automatically (i.e., without relying on user intervention) upon failure of the
exhaust air system to prevent sustained air positive pressurization of the
containment zone and to prevent the release of infectious material and
toxins. Examples of HVAC system interlocks include control logic built
into the containment zones building automation system and hard-wired
connections between the supply air and exhaust air fans.
3.5.10
Filtering exhaust air before recirculation or directly venting exhaust air

to the outdoors protects against infectious aerosols or aerosolized toxins
from being recirculated within the containment zone or building.
111
Requirement
Number(s)
Explanatory Note
3.5.11
Filtering exhaust air protects against airborne infectious material,

infectious aerosols, or aerosolized toxins from being recirculated within
the containment zone or building. HEPA ltration of the exhaust air adds
a higher level of protection to prevent the release of infectious material
and toxins from the containment zone.
3.5.12
Filtering exhaust air protects against airborne infectious material, infectious

aerosols, or aerosolized toxins from being recirculated within the
containment zone or building. HEPA ltration of the exhaust air adds a
higher level of protection to prevent the release of infectious material and
toxins from the containment zone. Including two stages of HEPA ltration
provides backup protection against the release of RG4 pathogens.
3.5.13
Certied HEPA lters are factory manufactured and tested in accordance

with the applicable IEST standard to demonstrate that they meet their
design function.
3.5.14
HEPA lter housings designed to withstand the pressures associated

with regular operation, gaseous decontamination, and the applied
pressures associated with routine integrity testing of containment zone
ductwork (i.e., pressure decay testing), reduce the potential of rupture
or developing a leak through which infectious material and toxins could
be released.
3.5.15
A mechanism to isolate HEPA lters (i.e., isolation dampers such as gas

tight damper, bubble tight damper) is critical to prevent the release of
contaminated air when removing, decontaminating, or testing (e.g., in situ
scan testing) lters.
3.5.16
An airtight seal on containment zone supply air ductwork prevents

the release of infectious material and toxins and facilitates gaseous
decontamination.
3.5.17
An airtight seal on containment zone exhaust air ductwork prevents

the release of infectious material and toxins and facilitates gaseous
decontamination.
112
Requirement
Number(s)
Explanatory Note
3.5.18
Location of airow control devices is critical for the effective control of

HVAC systems. This can be best achieved by locating airow control
devices on the supply air system on ductwork outside the containment
barrier, upstream of the supply isolation backdraft damper or HEPA
lter; and by locating airow control devices on exhaust air systems on
ductwork outside the containment barrier, downstream of the exhaust
HEPA lter.
3.5.19
Sections of the air supply and exhaust systems, including ductwork

and fans, outside the containment barrier require regular repairs,
maintenance, cleaning, and inspections to conrm that the HVAC
system continues to operate as designed. This is best achieved by
locating these sections of the air supply and exhaust systems so that they
are easily accessible by maintenance and other personnel.
3.6.1
Stand-off fasteners (e.g., surface mounted brackets) prevent the

accumulation of contamination (e.g., bedding, contaminated liquids)
behind conduits, piping, and other services by allowing accessibility for
maintenance, cleaning, and decontamination.
3.6.2
Shut-off valves and other controls for the main water supply are
located outside the containment zone to provide better accessibility
to maintenance and other personnel for regular maintenance, repair,
inspection, and emergency shut-off.
3.6.3
Backow prevention (e.g., reduced pressure principle systems, dual

check valves) and isolation valves (e.g., gate valves) on containment
zone water supply piping prevent potentially contaminated water or air
from entering water supply piping and allow control of water supply for
emergency shut-off.
3.6.4
Handwashing prevents the spread of many types of pathogens and

toxins outside of the containment zone. Locating a dedicated sink
near the point(s) of exit from the containment zone reduces the risk of
recontaminating hands after washing but before exit. Designating a
laboratory sink for handwashing is an alternate example to facilitate
handwashing by personnel upon exit.
113
Requirement
Number(s)
Explanatory Note
3.6.5
Handwashing sinks with hands-free capability (e.g., electronic eyes/

infrared sensors, foot pedals/pumps, elbow-controlled taps) reduce
contamination of the sink area and the potential for recontaminating
washed hands.
3.6.6
Emergency eyewash and shower equipment provide on-the-spot

treatment to ush out, dilute, and remove any hazardous materials,
including infectious material or toxins, that have contaminated the eyes,
face, or body before serious injury can occur.
3.6.7
Design features, such as capped or raised oor drains, are incorporated

into large scale production areas to prevent the inadvertent release of
infectious material or toxins into sanitary sewers.
3.6.8
Dikes or dams are used in areas where large scale process equipment
is kept to contain the full volume of large scale process uids in the event
of a leak or a spill and prevent the release of infectious material and
toxins to the sanitary sewer.
3.6.9
Drainage traps create a water seal that prevents contaminated air within
the containment zone from entering the piping, sewer, and/or efuent
decontamination systems. Deep seal P-traps with a seal depth of 125
mm or greater is a common example used in containment zones where
negative air pressure differentials are present.
3.6.10
Independent drain lines connected directly to an efuent decontamination

system prevent contaminated liquids from entering drain lines that service
other areas, or the sanitary sewer system.
3.6.11
The collection and redirection of autoclave condensate to drains

servicing areas inside the containment barrier prevent the release of
potentially contaminated liquids from the autoclave and directs them
to an efuent decontamination system so that they are decontaminated
prior to release to the municipal sewage system. Closed connections
prevent the release of the condensate before entering the drain; open
connections are only acceptable where the drain is located inside the
containment barrier or when the autoclave is equipped with an autodecontamination function that will effectively decontaminate the steam
condensate internally.
114
Requirement
Number(s)
Explanatory Note
3.6.12
Plumbing vent lines that are HEPA ltered or independent from those
of lower levels of containment prevent the release of infectious material
and toxins into the piping that services other areas.
3.6.13
Plumbing vent lines that are HEPA ltered prevent the release of
infectious material and toxins into the piping that service other areas.
Isolation dampers (i.e., bubble-tight or gas-tight) or other means of
properly isolating lters, allow the lters to be isolated for gaseous
decontamination.
3.6.14
Plumbing vent lines that are HEPA ltered and independent from those of
lower levels of containment provide backup mechanisms to prevent the
release of infectious material and toxins from CL4 into the piping that
service other areas. Isolation dampers (i.e., bubble-tight or gas-tight) or
other means of properly isolating lters, allow the lters to be isolated
for gaseous decontamination.
3.6.15
For personnel safety, supplied breathing air and hose connections are
provided in all areas of CL4 zones where positive-pressure suits are
worn, including chemical showers and suit change rooms.
3.6.16
In the event that the supplied breathing air system fails, backup air
supply systems (e.g., backup air cylinders, reserve air tank) need
to provide enough air to allow time for emergency evacuation of all
personnel from CL4 zones where positive-pressure suits are worn.
3.6.17
Locating light ballasts and starters outside the containment barrier makes
them readily accessible to personnel for maintenance and repair.
3.6.18
The continued operation of equipment critical for the containment and

security of infectious material and toxins (e.g., BSCs, ventilated cage
racks, electronic controlled access systems) during power outages is
crucial to maintain containment integrity and to safeguard the security
of the zone. In high containment zones, this includes HVAC systems
and controls, as well as equipment essential for personnel safety (e.g.,
air supply). Emergency power can be provided through the building
generator or a UPS system.
115
Requirement
Number(s)
Explanatory Note
3.6.19
In situations where a delay may occur before emergency power is

supplied, UPS allows the continued operation of life safety equipment
(e.g., supplied air to positive-pressure suits), building automation
systems, and security systems (e.g., electronic controlled access systems,
closed circuit television).
3.7.1
When properly maintained and used in conjunction with good

laboratory techniques, primary containment devices provide effective
personnel and environmental protection when working with open
vessels of infectious material, toxins, or infected animals. Examples
of primary containment devices include BSCs, fermenters, primary
containment caging (e.g., microisolator cages), ventilated cage racks,
and centrifuges with sealable cups or rotors.
3.7.2
When properly maintained and used in conjunction with good

laboratory techniques, primary containment devices provide effective
personnel and environmental protection when working with open
vessels of infectious material, toxins, or infected animals. Examples
of primary containment devices include BSCs, fermenters, primary
containment caging (e.g., microisolator cages), ventilated cage racks,
and centrifuges with sealable cups or rotors.
3.7.3
In the event of an exhaust fan failure, Class II B2 BSCs can produce a

reversal of airow from the face of the BSC (i.e., puff-back) as a result
of a delayed reaction to shutdown. Every effort should be made to
eliminate puff-backs mechanically (e.g., supply blower brake, isolation
damper for BSC supply intake). Where puff-backs cannot be eliminated,
it can be physically minimized (i.e., duration and airow velocity as
low as achievable) and additional operational mechanisms can be
implemented to address the risks associated with the puff-back, based
on the pathogen(s) and procedure(s) in use. Examples of operational
mechanisms include the use of additional PPE, such as respirators and
face protection, by all personnel in the immediate work area, and posting
emergency protocols to be followed in the event puff-back occurs.
116
Requirement
Number(s)
Explanatory Note
3.7.4
Preventing the release of infectious material and toxins from process

equipment, closed systems, and other primary containment devices
prevents contamination and protects personnel. This may include the use
HEPA lters on ports and vents, incineration, gaseous decontamination,
or fully enclosing the primary vessels in ventilated housings that are
exhausted through HEPA lters (e.g., walk-in containment enclosure).
3.7.5
Sensing devices (e.g., to monitor pressure, temperature) continually

monitor the integrity of process equipment and closed systems
during large scale processes and production, allowing failures to
be immediately signaled to personnel by audible or visual alarms in
order to initiate emergency procedures or repairs before a breach of
containment occurs.
3.7.6
The protective air curtain created at the front of the BSC is quite fragile
and can be easily disrupted by air currents or drafts created by trafc
or HVAC systems in close proximity. Locating a BSC away from high
trafc areas, doors, open windows, and air supply/exhaust diffusers
protects the BSC air curtain and protects personnel from exposure and
the release of pathogens and toxins.
3.7.7
The construction and installation of large pieces of equipment used

in large scale processes and production in a way that allows for
an effective means of cleaning and decontamination (e.g., autodecontamination or sterilization cycle) reduces the risk of exposure to the
3.7.8
Primary containment caging systems are designed to prevent the release

of pathogens, infectious material, or toxins shed by infected animals or
animals that have been intentionally exposed to a pathogen or toxin.
Ventilated microisolator cages are an example of animal caging that
could be used in an SA zone.
117
Requirement
Number(s)
Explanatory Note
3.7.9
Primary containment caging systems with HEPA ltration either on the

individual cage (e.g., lter-top cage) or through a ventilated enclosure
(e.g., microisolator cages in a HEPA ventilated cage rack) are designed
to protect personnel and prevent the release of pathogens, infectious
material, or toxins shed by infected animals or animals that have been
intentionally exposed to a pathogen or toxin.
3.7.10
Preventing animal escape is fundamental to protecting personnel and

animal safety, as well as preventing the release of infectious material or
infected animals.
3.7.11
The decontamination of waste and other contaminated material

inside the containment zone or their safe and secure transport to a
decontamination area (e.g., centralized facility decontamination
location or certied off-site waste disposal service) prevent the release
of pathogens from the containment zone. Examples of decontamination
technologies include autoclaves and incinerators.
3.7.12
Decontamination technologies (e.g., autoclaves, incinerator) within the

containment barrier allow for the decontamination of materials prior to
their removal from the zone.
3.7.13
Decontamination technologies (e.g., double-door autoclave, dunk tanks,

fumigation chamber) provided on the containment barrier allow for the
decontamination of all materials prior to removal from the containment
zone, and eliminates the possibility of the material becoming
recontaminated before leaving the zone.
3.7.14
Monitoring of operational parameters such as date, cycle number,

time, temperature, chemical concentration, and pressure conrms that
the decontamination technology is working properly (i.e., verifying that
parameters are achieving validated levels).
3.7.15
The appropriate temperature and treatment time to effectively

decontaminate waste materials are determined based on pathogen(s)
in use. For example, an autoclave capable of operating at 134C is
critical for the effective decontamination of all waste materials coming
from a containment zone where prions are handled and autoclaving is
used as a single-step decontamination process of this material.
118
Requirement
Number(s)
Explanatory Note
3.7.16
Chemical
decontamination
technologies
(e.g.,
chemical
decontamination showers, dunk tanks, efuent decontamination
systems) rely on the supply of chemicals for effective decontamination.
Maintaining supply tanks of these chemicals at a sufcient level is critical
for effective decontamination and to prevent a breach of containment.
3.7.17
Vacuum systems are used to aspirate liquid from a tube or to create a

void in ltration units (e.g., centralized vacuum system). Vacuum pumps
may cause aerosolization of infectious material or toxins and lead to
contamination of vacuum lines and pumps. A device such as a HEPA
lter, a small in-line lter (e.g., 0.2 m lter), or a disinfectant trap can
be used to protect vacuum systems from internal contamination with
3.7.18
A communication system increases personnel safety in the event of an

emergency in the containment zone, and can also be used to minimize
the movement of notebooks/papers and personnel into and out of the
containment zone. Examples of communication systems include telephones,
fax machines, intercom systems, networked computers, pagers, 2-way
radios, and communicating through a window (e.g., notes and signs,
hand signals).
3.7.19
Visual monitoring of laboratory work areas, animal zones, and large

scale production areas from outside the containment barrier improves
personnel safety and allows quick emergency response and assistance.
3.8.1
An efuent decontamination system allows all liquid waste leaving the

containment zone to be decontaminated, prior to release into sanitary
sewers. Efuent decontamination systems can include holding tank(s)
in conjunction with one or more decontamination tanks designed to
decontaminate liquid efuent waste by heat or chemical means in a
continuous ow or in static volumes (batches).
3.8.2
The appropriate parameters, such as temperature, chemical concentration,

and treatment time, to effectively decontaminate pathogens in liquid efuent
using a heat-based or chemical-based efuent decontamination system are
determined based on the pathogen(s) in use. For example, a temperaturebased efuent decontamination system capable of operating at 134C
is critical for the effective single-step thermal decontamination of all liquid
waste coming from a containment zone where prions are handled.
119
Requirement
Number(s)
Explanatory Note
3.8.3
Drain piping sloping towards the efuent decontamination system allows

for gravity ow and reduces the risk of blockage.
3.8.4
To be effective, it is essential that the entire efuent decontamination

system (i.e., piping, joints, valves, and tanks) be able to withstand
repeated exposure to the heat and/or the caustic chemicals necessary
for decontamination during normal operation. For example, chemical/
heat fused or welded joints protect the integrity of the entire system, as
conrmed by pressure decay testing.
3.8.5
Rooms housing an efuent decontamination system that serve as a

primary decontamination technology are designed to provide the
appropriate level of security to prevent unauthorized access, and in
the event of an efuent decontamination system failure, to prevent a
containment breach by containing any untreated waste.
3.8.6
Rooms housing an efuent decontamination system that serve as a

primary decontamination technology are designed to provide the
appropriate level of security to prevent unauthorized access, to contain
any infectious aerosols or aerosolized toxins, and in the event of an
efuent decontamination system failure, to prevent a containment breach
by containing any untreated waste.
3.8.7
Sludge and sediments can form blockages inside the efuent

decontamination system that are difcult to effectively decontaminate,
and impede the proper mechanics of the system. Examples of suitable
mechanisms include a grinder, trap, strainer, sloping of the piping, or
SOPs for routine maintenance procedures to deal with the sludge and
sediment buildup.
3.8.8
Alarm systems (e.g., audible alarm, automated electronic messaging) to

indicate failure(s) of the efuent decontamination system allow personnel
to quickly respond to a potential issue or emergency situation.
3.8.9
Temperature monitoring devices (e.g., electronic gauges, unit mounted

sensors) are essential to verify that the efuent decontamination system
reaches and maintains the required temperature for decontamination
of the infectious material or toxins. Annual calibration of temperature
monitoring devices conrms accuracy and functioning of the devices.
120
Requirement
Number(s)
Explanatory Note
3.8.10
Accurate labelling of all drainage piping leading to an efuent

decontamination system allows for the correct identication of these
components and a faster response by personnel in the event of a failure
or leak. Examples of suitable labelling include colour-coding, directional
arrows and hazard symbols.
3.8.11
Readily accessible drainage piping leading to an efuent

decontamination system allows maintenance and other personnel access
for regular maintenance, repair, and inspection.
3.8.12
HEPA lters provided on plumbing vent lines from the efuent

decontamination system prevent the release of pathogens and toxins
and the potential downstream contamination of the vent lines.
3.8.13
HEPA lters provided on plumbing vent lines from the efuent

decontamination system prevent the release of pathogens and toxins and
the potential downstream contamination of the vent lines. Two stages of
HEPA ltration provide backup protection against RG4 pathogens.
4.1.1
A robust biosafety program allows for the effective implementation

and maintenance of biosafety practices in accordance with the
organizations activities and regulatory requirements.
4.1.2
The biosafety representative(s) (i.e., designated BSO in licensed

facilities) is responsible for ensuring that the key elements of the
biosafety program are implemented to promote the safety and security
of the pathogens, toxins, and other regulated infectious material. In
order to effectively accomplish these responsibilities, the BSO requires
knowledge and experience appropriate for the facility and containment
level, and the pathogens and toxins handled.
4.1.3
Keeping contact information current prevents any delays in

correspondence, should information need to be relayed to the facility.
4.1.4
Program intent describes the scope of a facilitys activities, including the

pathogens, toxins, and other regulated infectious material, as well as
animal species in use.
121
Requirement
Number(s)
Explanatory Note
4.1.5
Program intent describes the scope of a facilitys activities, including

the pathogens, toxins, and other regulated infectious material, as well
as animal species in use. In containment zones where non-indigenous
animal pathogens are handled, submitting changes to program intent
or to SOPs that may impact biocontainment or biosafety to the CFIA
prior to implementation allows the CFIA to conrm the changes are
acceptable and that containment will be maintained.
4.1.6
An overarching risk assessment is a broad assessment that reects

the overall activities being conducted within the facility and is often
performed during the initial development or review of a facilitys
biosafety program.
4.1.7
A biosecurity risk assessment identies and prioritizes assets, denes

threats, and determines risk and mitigation strategies. It is through a
biosecurity risk assessment that it is determined whether it is necessary to
develop a basic or a complex biosecurity plan.
4.1.8
LRAs are site-specic risk assessments that are conducted to identify

hazards based on the infectious material and toxins in use and the
activities being performed. They are used to dene safe work practices
for the activities being performed and to develop SOPs.
4.1.9
A training needs assessment will determine key components of

the training program, including objectives, content, target groups,
implementation strategy, and retraining cycles.
4.1.10
The Biosafety Manual contains information related to the containment

zone. Individual elements of the Biosafety Manual may or may not be
housed in a single physical location. For example, in high containment
zones, training records may be stored outside the containment zone,
while printed copies of SOPs are generally kept inside the containment
zone. The Biosafety Manual can be developed using expertise from
various individuals such as the BSO, research or technical staff and a
medical advisor, if necessary.
122
Requirement
Number(s)
Explanatory Note
4.1.11
A description of the biosecurity plan constitutes one of the elements in

the Biosafety Manual. Evaluation and improvement of the biosecurity
plan promotes the continued renement of the overall biosafety
program, and may be done following an incident, changes in program
intent (e.g., working with a new pathogen), or any other situation that
may affect the biosecurity plan (e.g., facility renovations).
4.1.12
A medical surveillance program helps prevent, detect, and treat illnesses

associated with the exposure of laboratory personnel to pathogens and
toxins. The focus of the program is primarily preventive, although it also
provides a response mechanism through which a potential infection or
intoxication can be identied and treated in a timely manner.
4.1.13
LRAs and the requirements set out in other provincial/territorial and

federal legislation determine when respiratory protection is needed.
4.1.14
The training program includes topics such as the Biosafety Manual,

SOPs, entry/exit procedures, emergency response, and PPE. Evaluation
and improvement of the training program is critical for accuracy and
relevancy and to provide staff with training appropriate to their work.
4.1.15
SOPs are the foundation for safe work practices and describe the
specic chain of events for a particular activity.
4.1.16
A comprehensive ERP allows personnel to react quickly and

appropriately in a predetermined manner to emergency situations. It
sets out the procedures that personnel need to follow in response to
various emergency situations, in order to protect their health and safety,
to prevent the release of pathogens and toxins from the containment
zone, as well as to protect the security of the biological material stored
within the zone.
123
Requirement
Number(s)
Explanatory Note
4.2.1
Due to the increased risks associated with RG4 pathogens, establishing

a liaison with the local hospital or health care facility allows medical
staff to be aware of the pathogens that are handled or stored in the
containment zone, so that they can establish appropriate procedures for
treatment in the event of an accidental exposure.
4.2.2
Informing the appropriate internal personnel or authority promptly of

any potential exposure incidents or disease that is potentially related
to an exposure in the containment zone allows for rapid initiation of
the facilitys internal investigation process to determine if an exposure
(i.e., inhalation, ingestion, inoculation, or absorption of a pathogen
or toxin) has occurred. This will allow for timely follow up actions to
be taken to reduce or mitigate any further impacts including prompt:
identication and implementation of corrective actions to prevent other
potential exposures or further spread; administration of the appropriate
post-exposure and medical treatment; and notication of the PHAC of
the exposure incident.
4.2.3
Due to the high virulence of RG4 pathogens, immediate investigation of

unexpected work absences of CL4 personnel allows for prompt medical
attention to be obtained should the absence be related to a possible
exposure incident.
4.2.4
An emergency medical contact card provided by the employer

summarizes important information regarding higher risk pathogens or
toxins, as determined by an LRA, or the risk of exposure to Macacine
herpesvirus 1 (which is carried by certain non-human primates, such as
macaques), where non-human primates are being handled. This card
can provide information on pathogens that cause diseases that are
unlikely to be recognized by health care staff, information on routes of
transmission, symptoms, and preventive and therapeutic treatments.
124
Requirement
Number(s)
Explanatory Note
4.2.5
An emergency medical contact card provided by the employer

summarizes important information regarding the RG4 pathogens, and
any other infectious material handled, as determined by an LRA, or the
risk of exposure to Macacine herpesvirus 1 (which is carried by certain
non-human primates, such as macaques), where non-human primates
are being handled. This card can provide information on pathogens
that cause diseases that are unlikely to be recognized by health care
staff, information on routes of transmission, symptoms, and preventive
and therapeutic treatments.
4.3.1
The Biosafety Manual contains information essential for promoting a

safe and secure workplace. Training on its relevant aspects, including
associated SOPs, is critical in the understanding of its contents by
trainees and personnel.
4.3.2
Training personnel on the potential hazards and mitigation strategies

associated with the infectious material or toxins in use helps prevent
exposures and the release of contaminated materials. Providing training
on general signs and symptoms of concern helps staff quickly identify
potential exposure incidents.
4.3.3
In order to reduce the risk of personnel exposure and to maintain

containment of infectious material and toxins, personnel need to
understand the operation and design of containment zone systems
relevant to their activities. This may include the detection of system failures
and determining when it is safe, or not, to enter the containment zone.
4.3.4
Training on the safe use of laboratory equipment, including BSCs and other
primary containment devices, increases the likelihood that the devices will
be utilized correctly and increases personnel safety from exposures.
4.3.5
Since animals can behave unpredictably, and different species have

different risks, providing training in animal restraint and handling
techniques increases both personnel and animal safety, especially when
the animal is diseased or has been exposed to a pathogen or toxin.
125
Requirement
Number(s)
Explanatory Note
4.3.6
Accompanying and training individuals requiring temporary access

to the containment zone conrms and encourages the correct use of
procedures. This helps protect the individuals against exposure and
helps prevent breaches of containment.
4.3.7
Having personnel that are knowledgeable of the SOPs used in the

containment zone and procient in their use reduces the potential for
exposure or release when infectious material or toxins are being handled.
4.3.8
To protect all personnel from exposure and to help prevent breaches of

containment, the direct supervision of trainees by a trained individual
conrms that correct procedures are followed when handling infectious
material and toxins or when responding to an incident or accident.
4.3.9
Trained and knowledgeable staff creates a safer and more secure work
environment and help prevent breaches of containment. Reviewing the
training needs assessment on an annual basis will identify gaps in the
current training program. These gaps can be addressed by additional
or refresher training.
4.3.10
Annual refresher training on emergency response procedures conrms that

personnel remain knowledgeable on these infrequently used procedures
and that they can respond immediately and effectively to emergencies.
4.4.1
Using, wearing and storing dedicated PPE, such as lab coats, aprons,
solid-front gowns, coveralls, full body suits, and disposable sleeves, in
a specied area (e.g., CL3 laboratory, CL2 animal cubicle, or CL3
PM room) protects individuals from exposure and prevents the spread of
contamination outside of the containment zone.
4.4.2
Wearing eye or face protection (e.g., goggles or face shield) protects

personnel from exposure by preventing infectious material and toxins
from coming into contact with the mucous membranes of the eyes, and
where full face protection is worn, the nose, or mouth.
126
Requirement
Number(s)
Explanatory Note
4.4.3
Wearing dedicated footwear (e.g., boots or shoes) or additional

protective footwear (e.g., boot or shoe covers) at all times in animal
rooms, animal cubicles, and PM rooms protects individuals from
exposure by preventing the spread of contamination outside of the
containment zone.
4.4.4
Wearing appropriate gloves (e.g., latex, nitrile, vinyl) protects personnel

from exposure and prevents the spread of contamination by protecting
hands from contamination.
4.4.5
Full body coverage dedicated protective clothing (e.g., protective

suits, coveralls, surgical scrubs in conjunction with lab coat or back
closing gown) protects personnel from exposure by providing a layer
of protection between personnel and the infectious material and toxins.
4.4.6
Dedicated protective clothing and shoes (e.g., full body suits, coveralls,
surgical scrubs in combination with dedicated lab coat or back
closing gown), that are worn only inside the containment barrier (i.e.,
completely removed upon exit), prevent the spread of contamination
and protect against the release of infectious material or toxins from the
containment zone.
4.4.7
An additional layer of protective clothing (e.g., solid-front gowns with

tight-tting wrists, waterproof aprons, head covers) protects personnel from
exposure by providing an additional layer of protection in the event that
the outer layer of protective clothing is compromised or contaminated.
4.4.8
In CL4 zones where work is not performed entirely in a Class III BSC
line, positive-pressure suits that function properly (as determined by
integrity testing) protect personnel from exposure by providing an
additional layer of protection between personnel and the infectious
material and toxins, as well as delivering clean, breathable air.
4.4.9
Respirators protect personnel from airborne pathogens, infectious

aerosols, or aerosolized toxins that are not contained in a primary
containment device (e.g., a certied BSC or HEPA-ltered cage).
127
Requirement
Number(s)
Explanatory Note
4.5.1
Doors are kept closed to maintain the integrity of the containment barrier,
to maintain security, and to prevent animal escape. In high containment
zones, keeping doors closed is also essential for the proper operation of
the HVAC system.
4.5.2
Limiting access to the containment zone, animal rooms, animal cubicles,

and PM rooms to authorized personnel protects the safety of individuals
entering the containment zone and the security of all biological material
in the zone.
4.5.3
Due to the increased biosecurity risks associated with SSBAs, only

individuals having a valid HPTA 4ecurity $learance issued by the PHAC
are allowed to enter the part of a the facility where SSBAs are handled
and stored. A HPTA 4ecurity $learance is not necessary when an
individual is accompanied and supervised, or when the SSBAs are not
present or are locked up and inaccessible.
4.5.4
Due to the increased biosecurity risk associated with SSBAs and to

limit the number of personnel able to access the SSBA, only individuals
requiring access to the part of the facility where SSBAs are handled and
stored are allowed in that area.
4.5.5
Limiting access to mechanical and electrical services supporting the

containment zone (e.g., electrical panels, mechanical penthouse, and
HVAC system control areas) to authorized personnel protects the safety
of individuals entering these areas and the security of all biological
material in these areas and the containment zone.
4.5.6
Limiting access to mechanical and electrical services supporting the

containment zone (e.g., water supply shut-off valves and controls,
mechanical penthouse, electrical panels, and HVAC system control
areas) to authorized personnel protects the safety of individuals entering
these areas and the security of all biological material in these areas and
4.5.7
Limiting access to rooms housing an efuent decontamination system

to authorized personnel protects the safety of individuals entering these
areas and protects against the release of pathogens and toxins.
128
Requirement
Number(s)
Explanatory Note
4.5.8
Posting entry requirements at the point(s) of entry keeps individuals

entering the containment zone, animal room, animal cubicle, or PM
room informed of special entry requirements (e.g., PPE or additional
safety requirements that may exist depending on the nature of the
activities being conducted).
4.5.9
Verication of the correct readings on the airow monitoring devices

allows personnel to conrm that proper airow is provided before they
enter the area.
4.5.10
Storing personal clothing separate from dedicated PPE (including

protective clothing, where worn), either outside the containment barrier
or in dedicated change areas, protects individuals from exposure by
preventing cross-contamination to personal clothing and the spread of
contamination outside of the containment barrier.
4.5.11
Keeping personal belongings (e.g., outerwear, backpacks, purses, cell

phones, mp3 players) out of areas where infectious material or toxins
are handled or stored protects individuals from exposure by preventing
the contamination of these items and the spread of contamination
outside of the containment barrier. This also avoids the need to subject
personal belongings to the destructive effects of the decontamination
process in the event it becomes contaminated.
4.5.12
Leaving personal belongings (e.g., cell phones, mp3 players, purses,

outerwear) outside the containment zone or in assigned change areas
outside the containment barrier protects individuals from exposure
by preventing the contamination of these items and the spread of
contamination outside of the containment barrier. This also avoids the
need to subject personal belongings to the destructive effects of the
decontamination process upon exit from the containment barrier.
4.5.13
Removing personal clothes/footwear and donning of dedicated

protective clothing, including dedicated protective footwear, protects
individuals from exposure by preventing the contamination of personal
clothing and footwear and the spread of contamination outside of the
129
Requirement
Number(s)
Explanatory Note
4.5.14
Removing dedicated PPE in a particular order (i.e., gloves last) or manner

that protects personnel from exposure by preventing the contamination of
skin and hair and by reducing the potential of creating aerosols.
4.5.15
Gloves are the last piece of PPE that is removed when exiting a
containment zone. Handwashing is performed after removing gloves,
immediately before leaving the containment zone so that hands are not
recontaminated before exit. Handwashing is one of the most effective
ways to protect personnel from potential exposure by preventing the
spread of many types of pathogens and toxins through the inadvertent
contamination of surfaces, objects, or other items that may be touched
outside of the containment zone.
4.5.16
Gloves are the last piece of PPE that is removed when exiting a
containment zone. Handwashing is performed after removing gloves,
at the containment barrier immediately before leaving the containment
zone, animal cubicle or PM room, so that hands are not recontaminated
before exit. Handwashing is one of the most effective ways to protect
personnel from potential exposure by preventing the spread of many
types of pathogens and toxins through the inadvertent contamination
of surfaces, objects, or other items that may be touched outside of the
containment zone.
4.5.17
Removing dedicated PPE or the additional layer of PPE (e.g., dedicated

scrubs, boots, coveralls, lab coats, aprons, gowns, full body suits,
shoe covers, head and face protection), when exiting the containment
barrier of animal cubicles, PM rooms, or the containment zone protects
individuals from exposure by preventing the spread of contamination
outside of the containment barrier.
4.5.18
Eyeglasses can become contaminated when worn inside the

containment zone. Decontamination of eyeglasses at the containment
barrier protects individuals from exposure by preventing the spread of
contamination outside of the containment barrier.
130
Requirement
Number(s)
Explanatory Note
4.5.19
Removing all clothing worn inside the containment barrier, including

undergarments, and showering out upon exit of the animal cubicle, PM
room, or containment zone prevents the spread of contamination outside
of the containment barrier and protects the community by preventing the
release of non-indigenous animal pathogens.
4.5.20
Removing all clothing worn inside the containment barrier, including

undergarments, and showering out upon exit of the animal cubicle, PM
room, or containment zone prevents the spread of contamination outside
of the containment barrier and protects the community by preventing the
release of pathogens and toxins.
4.5.21
Exit procedures from CL4 zones are particularly stringent given the
nature of RG4 pathogens. A chemical shower to remove potential
contamination from positive-pressure suits, followed by the removal of all
clothing worn inside the containment barrier, including undergarments,
and showering protects individuals from exposure and prevents the
spread of contamination outside of the containment barrier.
4.6.1
Prohibiting activities such as eating, drinking, applying cosmetics,

inserting ear buds, or inserting/removing contact lenses reduces the
risk of personnel exposure through the contact of mucous membranes
of the eyes, nose, ears and mouth with contaminated or potentially
contaminated items.
4.6.2
Restraining or covering hair (including beards) reduces the risk that it

becomes contaminated through accidental contact with gloved hands,
specimens, containers, equipment, or surfaces.
4.6.3
Completely enclosed footwear prevents exposure, contamination, or

injury to the feet. Non-slip footwear, with low or no heels, prevents
slipping or tripping and enhances personnel safety.
131
Requirement
Number(s)
Explanatory Note
4.6.4
Jewellery may become contaminated or impede personnel

decontamination (e.g., via handwashing, showering) if infectious
material and toxins become trapped between these items and the skin.
Jewellery may also interfere with PPE (e.g., tear gloves or positivepressure suits). This also avoids the need to subject jewellery to the
destructive effects of the decontamination process upon exit.
4.6.5
Oral pipetting, or mouth aspiration, of any substance, especially

infectious material or toxins is strictly prohibited in the containment zone
to prevent the ingestion or aspiration of infectious material or toxins.
4.6.6
Intact skin provides protection from infection and intoxication. Any

breach of the skin (e.g., scratch, cut, wound, rash) may provide a portal
of entry for pathogens and toxins, and needs to be protected with a
bandage or other suitable waterproof dressing.
4.6.7
Maintaining trafc ow patterns for the movement of personnel,

equipment, samples, and animals from areas of least contamination
(i.e., clean) to areas of greatest contamination (i.e., dirty) limits the
spread of contamination.
4.6.8
Separating spaces dedicated to report writing and other paperwork

from areas where infectious material or toxins are actively handled
protects against contamination of these areas and any associated
materials and supplies that cannot be easily decontaminated.
4.6.9
Needles, syringes, and other sharp objects can cause punctures or

needlestick injuries, and potentially result in the injection or inoculation
of personnel. Examples of suitable alternatives include safety-engineered
needles and plasticware.
4.6.10
Needles, syringes, and other sharp objects can cause punctures or

needlestick injuries, and potentially result in the injection or inoculation
of personnel. There is a high risk of exposure from a needlestick injury
when performing activities such as bending, shearing, re-capping, or
removing needles from syringes that contain pathogens or toxins.
132
Requirement
Number(s)
Explanatory Note
4.6.11
The regular decontamination of work surfaces with an appropriate

disinfectant or neutralizing chemical minimizes the spread of
contamination and protects personnel from inadvertent exposure to
pathogens and toxins.
4.6.12
Having access to appropriate emergency assistance during and after

regular work hours enhances the personal safety of containment zone
personnel in the event of an emergency situation.
4.6.13
Routine verication that IDA is being maintained is essential to identify

potential breaches of the containment barrier.
4.6.14
The integrity of primary containment devices is veried to conrm that

they are functioning as intended and to prevent personnel exposure to
pathogens or toxins resulting from a breach of containment.
4.6.15
Moving or repairing BSCs can damage seals or the HEPA lter, and
result in a breach of containment. Certifying BSCs in accordance
with applicable standards or specications (e.g., NSF/ANSI 49 or
manufacturers specications, as applicable) conrms the integrity of the
HEPA lter and seals, and that the BSC is functioning as specied to
prevent exposure of individuals and the release of infectious material
and toxins into the environment.
4.6.16
Daily verication of containment and life safety system such as IDA,

supply air, disinfectant level in the chemical shower, and efuent
decontamination systems, are critical to maintain containment and to
protect personnel working in the containment zone.
4.6.17
The integrity of positive-pressure suits is veried to conrm that they

are functioning as intended to prevent personnel exposure and deliver
clean, breathable air.
4.6.18
Good microbiological laboratory practices (i.e., including the use of PPE,

handwashing, disinfecting work areas, use of procedures that minimize
the creation of aerosols, and proper decontamination and disposal of
materials) protect containment zone personnel from exposure by reducing
the risk of cross-contamination and the spread of contamination.
133
Requirement
Number(s)
Explanatory Note
4.6.19
Handling pathogens, toxins, and other regulated infectious material

in containment zones that do not meet the minimum requirements can
result in personnel exposure incidents or the accidental release of the
4.6.20
Safe and secure storage of pathogens, toxins, and other regulated

infectious material is critical to prevent the release of infectious
material and toxins. Additional precautions are necessary for proper
identication and security of all pathogens, toxins, and other regulated
infectious material stored outside the containment zone.
4.6.21
Safe and secure storage of pathogens, toxins, and other regulated

infectious material is critical to prevent the release of infectious
material and toxins. Additional precautions are necessary for proper
identication and security of all infectious material and toxins stored
outside the containment zone. For higher risk pathogens and toxins,
this includes the additional precaution of storage in locked storage
equipment that is within an area with limited access.
4.6.22
Safe and secure storage of pathogens, toxins and other regulated

infectious material is critical to prevent the release of pathogens and
toxins. Additional precautions are necessary for proper identication
and security of all pathogens, toxins, and regulated infectious material
stored outside the containment zone. Given their dual-use potential,
SSBAs have more stringent requirements to safeguard their security.
Fridges, freezers, or cabinets that are xed in place by bolting to the
wall or oor so that they are not-movable, and that are locked with
a padlock, combination lock, or other means to restrict access, are
examples of safe and secure storage equipment for use with SSBAs.
4.6.23
Due to the inherent characteristics of RG4 pathogens (e.g., high

virulence and severity of associated diseases, ease of transmission,
available host range, impact of introduction or release) and the
associated higher level of risk, the security requirements for RG4
pathogens are more stringent, and cannot be stored outside the CL4
containment zone.
134
Requirement
Number(s)
Explanatory Note
4.6.24
BSCs provide effective primary containment while simultaneously

providing personnel and environmental protection from infectious aerosols
or aerosolized toxins.
4.6.25
BSCs provide effective primary containment while simultaneously

providing personnel and environmental protection from infectious aerosols
or aerosolized toxins. Given their higher risk, a BSC is used for all
activities involving open vessels of RG3 and RG4 pathogens or toxins.
4.6.26
Removing gloves before exiting the BSC is an example of a procedure

that protects personnel from exposure and prevents the spread of
contamination by preventing the inadvertent contamination of surfaces,
objects or other items that may be touched after exiting the BSC. In
containment zones where two pairs of gloves are worn when handling
infectious material or toxins, the outer pair is removed upon exit from the
BSC to avoid the spread of contamination.
4.6.27
Handwashing is one of the most effective ways to prevent the spread

of contamination by many types of pathogens and toxins and protects
personnel from potential exposure (e.g., from touching eyes or mouth) by
preventing the inadvertent contamination of surfaces, objects, or other
items that may be touched; this includes activities such as paperwork
or computer work, that does not directly involve the manipulation of
infectious material. In containment zones where two pairs of gloves
are worn when handling infectious material or toxins, the outer pair is
removed upon exit from the BSC, and hands are washed immediately
after the inner pair is removed.
4.6.28
Centrifugation can create aerosols. When centrifuging infectious

material or toxins where inhalation is the primary route of infection or
intoxication, sealed safety cups or rotors will contain any infectious
aerosols produced. Sealed safety cups and rotors are unloaded in a
BSC to protect individuals from exposure to any aerosolized material
and prevent the spread of contamination.
135
Requirement
Number(s)
Explanatory Note
4.6.29
Sealed safety cups (or rotors) for centrifugation prevents the release of
infectious aerosols and aerosolized toxins. The sealed safety cups (or
rotors) are unloaded in a BSC to protect individuals from exposure to
any aerosolized material and prevent the spread of contamination.
4.6.30
Open ames (e.g., Bunsen burner) and on-demand open ames (e.g.,
touch-plate microburner) can damage the BSC HEPA lter and can
create turbulence that disrupts airow patterns. Examples of suitable
non-ame alternatives include microincinerators or sterile disposable
inoculation loops.
4.6.31
Ensuring that procedures are in place to prevent leaks, spills or drops,

or to contain the infectious material and toxins when these events occur,
protects personnel from exposure incidents and prevent the accidental
release of pathogens and toxins.
4.6.32
Closed systems or other primary containment devices (e.g., fermenters,

processing vessels) for use with the large volumes associated with
large scale cultures provide effective primary containment and provide
personnel and environmental protection from infectious aerosols and
aerosolized toxins.
4.6.33
The release of infectious aerosols or aerosolized toxins and the

contamination of exposed surfaces can be prevented when collecting
samples, adding materials, and transferring culture uids containing
infectious material or toxins from one closed system to another by
working through appropriate sampling ports.
4.6.34
Since autologous cells (i.e., cells derived from an individuals own body)
are not recognized as foreign by the immune system, the experimental
infection of ones own cells may place the individual at a greater risk of
infection in the event of an exposure.
4.6.35
A clean, uncluttered work environment allows appropriate decontamination

of the containment zone. It also minimizes slipping, tripping, fall, and
collision hazards that could potentially lead to exposure incidents or the
spread of contamination. Storing excess materials outside the containment
zone also protects this material from becoming contaminated.
136
Requirement
Number(s)
Explanatory Note
4.6.36
Routine cleaning by trained cleaning staff reduces contamination. Due

to the increased security measures, access controls, and risks associated
with the pathogens and toxins in use, these tasks can be assigned to
containment zone personnel. Alternatively, where dedicated janitorial
staff is expected to perform these activities, authorization to access the
containment zone for individual janitorial personnel is provided only
upon the completion of all necessary biosafety training and issue of any
HPTA 4ecurity $learances, as applicable.
4.6.37
Preventing the entry/exit of rodents and insects is important to prevent

the inadvertent transfer and transport of pathogens and toxins out of the
containment zone. Examples of suitable measures include the installation
of traps, screens, or door sweeps, in conjunction with regular monitoring
and maintenance.
4.6.38
Water seals in drainage traps located in sinks, showers, and oor

drains are maintained through regular use or routine lling to prevent
the passage of potentially contaminated air and to limit the amount of
stagnant contaminated water.
4.6.39
Decontamination of HEPA lters through in situ fumigation with

formaldehyde or vaporized hydrogen peroxide allows for the
decontamination of HEPA lters prior to their removal. In containment
zones where prions are handled, an alternative mechanism for the
safe removal of HEPA lters is required since prions are not completely
inactivated when using the most common decontamination methods.
Examples of suitable alternatives include using HEPA lters with a bagin/bag-out capability or using procedures to contain the HEPA lter for
removal followed by its subsequent decontamination.
4.6.40
To avoid interruptions in operation, the availability of a basic tool kit

inside high containment zones allows containment zone personnel
or trained maintenance personnel to conduct minor repairs that do
not require the assistance of a specialized tradesperson. This also
avoids the need to subject the tools to the destructive effects of the
decontamination process.
137
Requirement
Number(s)
Explanatory Note
4.7.1
Since animals can behave unpredictably, and different species entail

different risks, using proper animal restraint and handling techniques
provides both personnel and animal safety, especially when the animal
is diseased or has been exposed to a pathogen or toxin.
4.7.2
Labelling of primary containment caging housing infected animals is

critical for personnel to be aware of the risks associated with the animals.
4.7.3
Animal handling procedures that minimize the creation of aerosols, the

dissemination of dust containing infectious material or toxins, and limit
the amount of allergens dispersed reduce the risk of personnel exposure
and prevent the spread of contamination.
4.7.4
When animal procedures that may lead to aerosolized or airborne

pathogens are performed (e.g., necropsy, surgery, inoculation),
appropriate procedures for entry to and exit from the animal cubicle or
PM room protect individuals from exposure and prevents the spread of
contamination. Examples include procedures to avoid the simultaneous
opening of doors in a sequence (e.g., anteroom doors) when entering
into an animal cubicle or PM room or allowing sufcient time for any
aerosols to settle before opening the door to enter/exit.
4.7.5
Live animals are securely moved into and around the containment zone
to prevent animal escape and injury. Safe and secure movement of
carcasses protects individuals from exposure by preventing the spread
of contamination.
4.7.6
Transporting animal carcasses in sealed, leakproof and impact-resistant

containers protects individuals from exposure by preventing the spread
of contamination. This is not required when transporting via the dirty
corridor, as the dirty corridor is considered contaminated.
4.7.7
Inoculation, surgical, and necropsy techniques generally require the

use of sharp objects and often have the potential for creating aerosols.
The appropriate choice of equipment and the use of skilful techniques
prevent punctures or needlestick injuries that could potentially result in
the injection or inoculation of personnel.
138
Requirement
Number(s)
Explanatory Note
4.7.8
Conducting inoculation, surgical, and necropsy procedures on smallsized animals in a certied BSC or other appropriate biocontainment
device, such as a ventilated animal surgery, downdraft, or backdraft
station, provides effective primary containment and protects personnel
and the environment from infectious aerosols or aerosolized toxins.
4.7.9
Decontamination and cleaning the skin and hair or feathers of the

animal following a procedure protect individuals from exposure and
prevents the spread of contamination.
4.8.1
Removing and separately treating gross contamination, especially

organic material, is essential to allow the effective decontamination of
surfaces and equipment.
4.8.2
Several factors inuence the effectiveness of a disinfectant, including

the type of pathogen being decontaminated (e.g., virus, bacteria,
prion), its state (e.g., vegetative or spore form), as well as the organic
load (i.e., the amount of organic material such as soil, bedding,
litter, feed, or manure present on a surface or in a suspension) of the
material being decontaminated. Neutralizing chemicals for microbial
toxins are selected based on the specic physical properties of
the particular toxin(s) in use, as there is a wide variety of microbial
toxins with considerably different characteristics. Disinfectants and
neutralizing chemicals available inside the containment zone allow for
decontamination of surfaces (e.g., benchtops and containers) and for
quick response to biological spills.
4.8.3
Containers that are leakproof, puncture resistant, and tted with lids
prevent punctures from sharp objects and exposure incidents for
containment zone personnel, as well as for individuals handling waste.
4.8.4
Decontamination of BSCs and primary containment devices prior to

maintenance protects individuals from exposure when working to maintain
these devices. It also prevents the release of pathogens and toxins.
139
Requirement
Number(s)
Explanatory Note
4.8.5
A known or suspected exposure (e.g., from a splash or spill) may lead

to the contamination of PPE and clothing. The decontamination of these
items following such events protects individuals from exposure and
prevents the spread and release of pathogens and toxins.
4.8.6
PPE, including protective clothing, may become contaminated during

use. Decontamination of PPE prior to disposal or laundering not only
protects personnel from exposure by preventing the inadvertent release of
pathogens or toxins, but also protects individuals handling the PPE (e.g.,
laundry service personnel) once it is removed from the containment zone.
4.8.7
Decontamination of contaminated liquids using a validated method

prior to discharge into sanitary sewers prevents the release of
pathogens or toxins.
4.8.8
Decontamination of contaminated material, including waste and

equipment, or the surface decontamination of transport containers,
protects against the inadvertent release of pathogens and toxins from the
containment zone, and protects the safety of individuals who handle,
clean, and dispose of these materials.
4.8.9
Decontamination of equipment, materials, waste, and the surface

decontamination of transport containers at the containment barrier, prior
to their removal from the containment zone, prevents the inadvertent
release of pathogens and toxins from high containment zones and
protects the safety of individuals who handle these materials. Properly
labelling the items as decontaminated prevents non-decontaminated
items from being inadvertently removed from the containment zone.
4.8.10
Validation of decontamination equipment and processes, when

changes in the process are made (e.g., new protocol, parameters,
or concentrations) or when new pathogens that have not previously
been handled by containment zone personnel, conrms that the
equipment is functioning properly and that the process is effective for the
decontamination of materials prior to their removal or release from the
containment zone.
140
Requirement
Number(s)
Explanatory Note
4.8.11
Verication of decontamination equipment (through the use of biological

indicators, chemical integrators, or parametric monitoring devices)
is performed upon initial use, and routinely thereafter to conrm that
the process is effective for the decontamination of materials prior to
their removal from the containment zone. An LRA will help determine
the procedures for routine monitoring, taking into consideration the
frequency of use of the equipment.
4.8.12
The simultaneous opening of both doors of a double-door barrier

autoclave or other pass-through technology will interfere with IDA and
may result in a breach of containment. Keeping at least one door closed
safeguards containment integrity and prevents the release of pathogens
and toxins.
4.8.13
Bedding from infected animals may be contaminated with infectious

material and toxins. The use of containment devices (e.g., BSC or a
ventilated cage changing station) when handling contaminated bedding
protects individuals from exposure and prevents the spread and release
of pathogens and toxins. Decontaminating the bedding while still inside
the containment cage can achieve the same goal.
4.8.14
Decontaminating animal cubicles, PM rooms, and the dirty corridor

whenever they are grossly contaminated and at the end of each
experiment protects personnel from exposure by preventing the spread
of pathogens and toxins.
4.8.15
A full room decontamination process is validated to conrm it is effective

against the pathogens and toxins handled. Full room decontamination
may be performed following a spill, at the end of a project, or to
facilitate regular preventive maintenance and performance testing.
4.9.1
A comprehensive ERP allows personnel to react quickly and

appropriately and in a predetermined manner to emergency situations.
It sets out the procedures for personnel to follow in response to various
emergency situations, in order to protect them from exposure, and to
prevent the release of pathogens and toxins from the containment zone.
141
Requirement
Number(s)
Explanatory Note
4.9.2
Including response procedures for emergencies involving infectious

material and toxins that are stored outside the containment zone allows
for the continued safe and secure storage of infectious material and
toxins, and also prevents the release of pathogens and toxins.
4.9.3
Including response procedures to describe emergency situations in a

CL4 zone that could potentially lead to a breach of containment, or loss
of breathing air, protects individuals from exposure to pathogens and
hazardous chemical decontaminants. Including response procedures
in the event of the loss of chemical decontamination shower protects
against the spread of contamination and the release of pathogens from
4.9.4
Including response procedures that describe emergency situations

where showers are necessary upon exit, protects against the spread
of contamination and the release of pathogens and toxins from the
containment zone.
4.9.5
Including response procedures for emergency exit to describe situations

where normal exit procedures (e.g., showers taken upon exit from
the containment barrier) are not followed during life-threatening
emergencies, protects against the spread of contamination and the
release of pathogens and toxins from the containment zone.
4.9.6
An incident (e.g., a splash or spill) may lead to the contamination of

oors and other surfaces. Decontamination following such events
protects individuals from exposure and prevents the spread and
release of pathogens and toxins. Having a biological spill kit inside the
containment zone, and readily available, is essential for a quick spill
response and to limit the spread of contamination.
4.9.7
Immediately reporting incidents to the appropriate internal authority

as established by the facilitys reporting chain allows for a rapid and
appropriate response to minimize the risk to personnel (e.g., administration
of medical assistance or rst-aid), to contain any possible release of
pathogens, toxins, and other regulated infectious material, to initiate
any repairs or corrective actions to containment systems, and, when
applicable, to initiate external notication to the regulatory agencies.
142
Requirement
Number(s)
Explanatory Note
4.9.8
An incident investigation determines the root cause(s) of the incident

(i.e., why it took place) and whether it was an isolated event, and may
allow new procedures to be implemented to prevent the recurrence of
similar events.
4.9.9
The submission of an exposure notication report without delay (i.e., as

soon as reasonably possible) allows the PHAC to assess the severity of
the exposure incident and assist the licence holder in their response, if
requested or necessary. It also prevents other potential exposures.
4.9.10
An exposure follow-up report updates the PHAC on the notied

exposure incident and allows for the identication, monitoring, and
analysis of trends related to exposures over time. New information is
included in the exposure follow-up report and may include the status
of the investigation, root cause analysis, risk mitigation strategies put in
place to prevent recurrence, and outcomes (if known).
4.10.1
Training records provide information on personnels participation in

training, and their successful completion of training requirements conrms
that employees have met all of the training requirements of their position.
4.10.2
An inventory is a list (or lists) of (biological) assets in the possession of the

containment zone. Inventories are basic records that allow individuals who
are accountable for the control of pathogens, toxins, and other regulated
infectious material in a containment zone/facility to manage and control
the material. By means of an inventory, a facility manager, containment
zone director, or BSO can document and identify the hazards, describe
the level of risk in a form readily accessible to personnel, and protect the
assets from being misused, misplaced, stolen, abandoned, or released.
Inventory information may be accessed in response to an emergency
situation where samples need to be relocated (e.g., power outage) or in
response to requests from the PHAC or the CFIA (e.g., targeted request for
specic pathogens, such as poliovirus stocks). Inventories can exist in many
different forms. The specic format (e.g., electronic or paper inventory, a
list, a logbook of samples) for establishing and maintaining inventories is
determined on an institutional/facility/containment zone level.
143
Requirement
Number(s)
Explanatory Note
4.10.3
An increased level of detail in the inventory is specied due to the

increased risk of deliberate theft associated with RG3 and RG4
pathogens and SSBA toxins so that containment zone personnel can
quickly determine if any material is missing. In addition to the exact
location (e.g., room, freezer, rack, box), inventories containing sufcient
detail to identify the specic pathogen or toxin, its risk group, the
number of vials and quantity contained in the vials, allow personnel
to be aware of the pathogens, toxins, and other regulated infectious
material that are stored long-term in the containment zone.
4.10.4
As-built drawings provide a record of all changes made to the

specications and working drawings during the construction process,
and show the exact dimensions and location of all elements of the
completed facility. In conjunction with physical specications, and
performance and verication test reports, they are a record that
demonstrates that the containment zone has been built to meet the
physical requirements.
4.10.5
Records, which may include formal internal inspection reports and followup notes from a Biosafety Committee or a Health and Safety Committee,
or informal notes/documentation on repairs required, are evidence that
internal inspections and follow-up actions have been performed.
4.10.6
Records, which can include documentation of BSC testing and

reports of certication in accordance with applicable standards and
specications, HEPA lter testing, equipment integrity testing, and
building maintenance, are evidence that internal inspections, repair, or
testing have been performed.
4.10.7
Calibration certicates are evidence that the instruments used for

performance and verication testing were accurate at the time of their
use. Calibration certicates from third-party certiers can be obtained
by request, where applicable.
4.10.8
Maintaining records of entry/exit of all individuals (including emergency

responders) conrms who was present in the containment zone in the
event of an emergency, for biosecurity purposes, and for historical
purposes in the event of known or suspected cases of exposure.
144
Requirement
Number(s)
Explanatory Note
4.10.9
Records of validation and routine testing of decontamination

technologies and processes provide evidence that the decontamination
methods are effective against the infectious material and toxins handled
or stored in the containment zone. Records of validation and verication
from third-party biohazardous waste disposal facilities can be obtained
by request, where applicable.
4.10.10
Maintaining records pertaining to licence activities and animal

pathogen import permit requirements for a minimum amount of time
allows for the documentation of pathogens, toxins, and other regulated
infectious material being, or that have been, handled or stored within
the containment zone. It also allows for their review by the PHAC or the
CFIA, if requested.
4.10.11
Records of incidents involving pathogens, toxins, other regulated

infectious material, or losses of containment allow for the monitoring
of trends and can serve to improve systems or procedures. Keeping
them on le for the specied periods allows for their review by the
PHAC or the CFIA, if required. Lessons learned and corrective actions
implemented in response to these reports are a useful reference in future
investigations and can also serve as learning tools.
4.10.12
Limiting access to sensitive information, including inventories, pathogen

and toxin storage locations, lists of authorized personnel, access codes,
and passwords, safeguards the security of the information and of the
5.1.1
Conducting regular performance and verication tests provides proof

that the containment systems are functioning as designed and intended
and that the minimal acceptable criteria continue to be met. Reports of
each test provide a record that demonstrates the appropriate test(s) have
been performed. Testing frequency is based on the risks associated
with equipment malfunctioning or failing, and is higher for higher
containment levels. Tests are performed following any changes, repairs,
or modications to any component that may affect the containment
system to conrm it continues to function as intended before operation
involving pathogens and toxins resumes.
145
Requirement
Number(s)
Explanatory Note
5.1.2
Faults or deterioration that may affect containment, including faulty

equipment, cracked or chipped paint or oors, and faulty lighting, may not
be noticed by personnel on a day-to-day basis. Faults and deterioration
are identied through routine visual inspections of the containment zone,
including surfaces, oors, walls, ceilings, and equipment.
5.1.3
Routine inspection and replacement of in-line lters is developed and

followed as part of a containment zone maintenance plan to verify
that lters continue to function as intended and to prevent a breach
of containment. This test is intended to verify the performance of lters
where present in accordance with requirements 3.5.4, 3.7.8, 3.7.9,
and 3.7.16.
5.1.4
Validation of decontamination technologies and processes conrms the

proper functioning of the equipment and the effective decontamination
of materials prior to their disposal or removal from the containment zone.
In cases where biological or chemical indicators are not appropriate
(i.e., prions), parametric monitoring devices can be used to accurately
monitor the performance of decontamination equipment. Examples
include thermocouples and gauges that capture cycle time, temperature,
and pressure. A thermocouple is an application-specic parametric
device for the validation of heat-based decontamination technologies
(e.g., autoclaves) when used for inactivating prions. This test is
intended to verify the performance of decontamination technologies in
accordance with requirements 3.7.11, 3.7.12, 3.7.13, and 4.8.10.
5.1.5
Testing of Class II BSCs to conrm they operate as intended prevents

personnel exposure resulting from a breach of containment. This test is
intended to verify the performance of Class II BSCs where present in
accordance with requirements 3.7.1, 3.7.2, 3.7.3, and 4.6.15.
5.1.6
Testing of BSCs and custom ventilated enclosures to conrm they

operate as intended prevents personnel exposure resulting from a
breach of containment. This test is intended to verify the performance
of BSCs and custom ventilated enclosures where present in accordance
with requirements 3.7.1, 3.7.2, 3.7.3, and 4.6.15.
146
Requirement
Number(s)
Explanatory Note
5.1.7
Testing of primary containment devices to conrm they operate as

intended prevents personnel exposure resulting from a breach of
containment. This test is intended to verify the performance of primary
containment devices, other than a BSC, where present in accordance
with requirements 3.7.1, 3.7.2, 3.7.4, 3.7.8, and 3.7.9.
5.2.1
Conducting regular performance and verication tests provides proof

5.2.2
Conducting regular performance and verIcation tests provides proof

5.2.3
Controlled access and security systems restrict access to the containment

zone(s) to authorized personnel only. Verication conrms that the
controlled access systems operate as designed, such that a correct
code, card, or biometric trait allows access and an incorrect code,
card, or biometric trait does not. Verication of other security systems
(e.g., closed circuit television) conrms that they operate as specied.
If key locks are used, verication can also include verifying that keys
are only distributed to authorized personnel and that they are nonreproducible. This test is intended to verify the performance of controlled
access and security systems, where installed in accordance with
requirements 3.3.4, 3.3.5, 3.3.6, 3.3.7, and 3.3.8.
147
Requirement
Number(s)
Explanatory Note
5.2.4
Containment systems used in high containment zones (including, but not

limited to, controls, fans, critical containment devices, communication
devices, and efuent decontamination systems) are critical for personnel
and environmental safety. Testing of the emergency power and UPS
systems under representative electrical load conditions conrms
that all critical containment systems will continue to operate during a
power outage. If live load testing is not possible, simulated load
testing is acceptable. This test is intended to verify the performance of
emergency power and UPS systems, where present in accordance with
requirements 3.6.18 and 3.6.19.
5.2.5
Communication systems allow the transmission of information in and out

of the containment zone, and are especially essential in the event of
an emergency. Verication of communication systems conrms that they
operate as intended. This test is intended to verify the performance of
communication systems installed in accordance with requirement 3.7.18.
5.2.6
Intact seals and surfaces maintain the containment barrier, and allow
areas or surfaces to be effectively decontaminated. A visual inspection
allows the identication of areas that have lost their integrity and that
need repair. A visual inspection includes verifying oors, walls, and
ceilings, as well as oor/wall and wall/ceiling joints for cracks, chips,
and wear. Identifying surfaces on the containment barrier that have
lost their integrity is essential to protect personnel from exposure and
to prevent a breach of containment. This test is intended to verify the
performance of seals and surfaces in accordance with requirements
3.2.6, 3.2.10, 3.2.11, 3.2.12, 3.4.1, 3.4.2, 3.4.4, 3.4.7, 3.4.8,
and 3.4.9 as applicable.
5.2.7
Visually demonstrating IDA at all critical doors on the containment barrier

will verify that air ows toward areas of higher containment, according
to design, and never the reverse. This test is intended to verify that IDA
is provided in accordance with requirements 3.5.1, 3.5.2, and 3.8.6.
5.2.8
Testing of HEPA lters conrms their integrity and prevents a breach of

containment and the release of infectious aerosols or aerosolized toxins
from the containment zone. This test is intended to verify the performance
of HEPA lters installed in accordance with requirements 3.5.6, 3.5.7,
3.5.8, 3.5.10, 3.5.11, 3.5.12, 3.6.12, 3.6.13, 3.6.14, 3.7.9,
3.8.12, and 3.8.13.
148
Requirement
Number(s)
Explanatory Note
5.2.9
Properly functioning door interlocks prevent the simultaneous opening

of two doors, which could lead to a breach of containment, while
properly functioning emergency overrides are critical in emergency
situations. Verication of door interlocks conrms that the interlocks
operate as designed, and that associated emergency manual overrides
release all interlocks when activated. Door interlocks can be tested by
opening the critical door and attempting to simultaneously open the
associated interlocked doors. Door interlocks are tested one at a time,
in both directions. This test is intended to verify the performance of door
interlocks and associated manual overrides installed in accordance with
requirements 3.2.7, 3.2.8, 3.2.9, 3.3.17, 3.3.18, and 3.3.19.
5.2.10
Testing of the low level alarm of chemical decontamination technologies

conrms that the alarm is triggered when the level of disinfectant drops
below the minimum set point. Testing of other alarms in decontamination
technologies conrms that they function as designed. This test is intended
to verify the performance of alarms on decontamination technologies
present in accordance with requirements 3.2.7, 3.2.8, 3.2.9, 3.7.16,
and 3.8.8.
5.2.11
Testing of water supply backow preventers (e.g., reduced pressure

backow preventers) conrms that they are working as intended and
protect the main water supply from contamination in the event of a
reversal of water from the containment facility. This test is intended to
verify the performance of backow prevention installed in accordance
with requirement 3.6.3.
5.2.12
Testing the integrity of seals of containment barrier penetrations,

animal cubicle penetrations, and PM room penetrations, including all
conduits and wiring, as well as seals around doors, windows, and
pass-through technologies including autoclaves and dunk tanks, with a
smoke pencil identies seals that have lost their integrity and that need
repair. Identifying surfaces on the containment barrier that have lost their
integrity is essential to protect personnel from exposure and to prevent
a breach of containment. This test is intended to verify the seals present
in accordance with requirements 3.2.6, 3.2.10, 3.2.11, and 3.2.12.
149
Requirement
Number(s)
Explanatory Note
5.2.13
Testing of compressed breathing air and associated systems, including

the switchover to backup system and alarm response, conrms that
they operate as intended and helps prevent personnel exposure. This
test is intended to verify the performance of compressed breathing air
and associated systems in accordance with requirements 3.6.15 and
3.6.16.
5.2.14
Testing of positive-pressure suits, including a visual inspection of seams,

closures, gaskets, joints and pressure testing by suit ination, conrms
that they operate as intended and helps prevent personnel exposure.
This test is intended to verify the integrity of positive-pressure suits worn
in the containment zone in accordance with requirement 4.4.8.
5.2.15
Testing of chemical decontamination shower systems in the containment

zone, including verication that the appropriate concentration or
volume of disinfectant is discharged, chemical level and conductivity
meters function as intended, and that the timer (if applicable) reaches
the minimum set time, conrms that they operate as designed. Testing of
the low level alarm conrms that the alarm is triggered when the level
of disinfectant drops below the minimum set point. This test is intended
to verify the performance of chemical decontamination shower systems
and associated parametric monitoring and recording devices and alarms
installed in accordance with requirements 3.3.16, 3.7.14, and 3.7.16.
5.3.1
Drainage piping is tested in accordance with the National Plumbing

Code of Canada to conrm that the system is working correctly. Testing
includes all drains and associated piping, as well as associated
vent lines connected to the efuent decontamination system. This
test is intended to verify the performance of drain piping installed in
accordance with requirement 3.6.10.
150
Requirement
Number(s)
Explanatory Note
5.3.2
Testing of HEPA lter housings conrms their integrity and prevents

a breach of containment and the release of infectious aerosols or
aerosolized toxins from the containment zone. This test is intended to
verify the integrity of HEPA lter housings installed in accordance with
requirement 3.5.14.
5.3.3
HVAC and HVAC control systems are critical for environmental safety
and personnel safety. Testing the HVAC system and HVAC system
components through a variety of failure scenarios demonstrates that
these systems can continue to prevent a breach of containment and the
release of infectious aerosols or aerosolized toxins from the containment
barrier, even in the event that one or more components of the system
fails. These tests are intended to verify the performance of the HVAC
system and HVAC system components and features in accordance with
requirements 3.5.1, 3.5.2, 3.5.3, 3.5.4, 3,5.5, 3.5.9, 3.5.18, and
3.7.3, as applicable.
5.3.4
Testing of containment zone ductwork conrms its integrity and prevents

a breach of containment and the release of infectious aerosols or
aerosolized toxins from the containment zone. This test is intended to
verify the integrity of the supply air and exhaust air ductwork installed
in accordance with requirements 3.5.7, 3.5.8, 3.5.16, and 3.5.17.
5.3.5
Integrity of the containment barrier in containment zones where doors

can be sealed (CL3-Ag and CL4) is veried by pressure decay testing.
Identifying surfaces on the containment barrier that have lost their
integrity is essential to protect personnel from exposure and to prevent
a breach of containment. This test is intended to verify the integrity of
the containment zone designed in accordance with requirements 3.2.6,
3.2.11, 3.2.12, 3.3.20, and 3.3.21.
151

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Canadian Biosafety Standard, Second Edition

is available on the Internet at the following address:

PDF Cat.: HP45-7/2015E-PDF

CHAPTER 2 HOW TO USE THE

Abbreviations, Definitions, and References. . . . . . . . . . . 14

CHAPTER 3 PHYSICAL CONTAINMENT

CHAPTER 4 OPERATIONAL PRACTICE

Biosafety Program Management . . . . . . . . . . . . . . . . . .

CHAPTER 5 PERFORMANCE AND VERIFICATION

Performance and Verification Tests for All

Human pathogens and toxins: Laboratory Biosafety Guidelines, 3rd Edition,

Terrestrial animal pathogens: Containment Standards for Veterinary Facilities,

Canadian Biosafety Standard (CBS) 2nd Edition

ABBREVIATIONS AND ACRONYMS

ABBREVIATIONS AND ACRONYMS

Agriculture (i.e., CL2-Ag, CL3-Ag)

Automated Import Reference System

American National Standards Institute

American Society of Mechanical Engineers

Biological safety cabinet

Biological safety ofcer

National Standard of Canada

Canadian Biosafety Handbook

Canadian Biosafety Standard

Canadian Biosafety Standards and Guidelines

Canadian Food Inspection Agency

Containment level (i.e., CL1, CL2, CL3, CL4)

Canadian Standards Association

Containment Standards for Facilities Handling Aquatic Animal Pathogens

Containment Standards for Facilities Handling Plant Pests

Emergency response plan

Health of Animals Act

Health of Animals Regulations

High efciency particulate air

Human Pathogens and Toxins Act

Canadian Biosafety Standard (CBS) 2nd Edition

Human Pathogens and Toxins Act 6ecurity &learance

Human Pathogens and Toxins Regulations

Heating, ventilation, and air conditioning

Inward directional airow

Institute of Environmental Sciences and Technology

Inches of water gauge (unit of pressure; 1 in. w.g. = 250 Pa)

Large animal containment zone

Local risk assessment

National Sanitation Foundation

Public Health Agency of Canada

Post mortem room

Personal protective equipment

Pathogen safety data sheet

Risk group (i.e., RG1, RG2, RG3, RG4)

Small animal containment zone

Sheet Metal and Air Conditioning Contractors National Association

Standard operating procedure

Security sensitive biological agent

UninterruptJCMe power supply

An unplanned event that results in injury, harm, or damage.

Dedicated room or adjoining rooms that are used for activities

A suspension of ne solid particles or liquid droplets in a gaseous

A pathogen that is capable of moving through or being carried

Doors that are designed to allow no leakage of air (0%) under