CCAC guidelines on: genetically-engineered animals used in science

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Dec 11, 2012 (4 years and 6 months ago)

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Draft


August 2011



FINAL DRAFT

August 2011






CCAC guidelines on:

genetically
-
engineered animals

used in science



Thank you for your willingness to review this CCAC draft document.



The deadline for comments
is October 28, 2011
.

Comments may be returned to the CCAC in
whatever format is convenient (typed or handwritten, and
written on the draft or listed separately, identifying the paragraph number where appropriate).

Comments should be sent to:

Ms. Julie Dale

Director of Guidelines Development

Canadian Council on Anim
al Care

1510
-
130 Albert Street


Draft


August 2011



Ottawa, ON K1P 5G4

email:
jdale@ccac.ca


Please note that it is
still
a draft document and therefore
may

undergo
further

change before publication
.




TABLE OF CONTENTS

1.

PREFACE

................................
................................
................................
................................
......

1

SUMMARY OF THE GUIDELINES LISTED IN THIS DOCUMENT

................................
................................

3

2.

GENERAL CONSIDERATIO
NS

................................
................................
................................
.........

9

2.1

Definition of genetically
-
engineered animal and scope of this guidelines document

..................

9

2.2

Ethical overview

................................
................................
................................
..........................

10

2.3

Communication

................................
................................
................................
...........................

11

2.4

Research planning

................................
................................
................................
.......................

12

3.

FACILITIES, FACILITY MANAGEM
ENT AND STAFFING

................................
................................
...

16

3.1

Facilities and facility management

................................
................................
..............................

16

3.2

Staffing

................................
................................
................................
................................
........

17

4.

GENERATION AND WELFARE ASSESSMENT

................................
................................
.................

19

4.1

Generation of a new genetically
-
engineered animal line

................................
...........................

19

4.2

Welfare
assessment

................................
................................
................................
....................

22

4.3

Monitoring and mitigation strategies

................................
................................
.........................

29

5.

ACQUISITION AND TRANSPORTATION

................................
................................
........................

30

5.1

Acquisition of existing genetically
-
engineered animal lines

................................
.......................

30

5.2

Transportation

................................
................................
................................
.............................

31

6.

HUSBA
NDRY

................................
................................
................................
..............................

33

6.1

Identification and record
-
keeping

................................
................................
...............................

33

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August 2011



6.2

Housing and husbandry

................................
................................
................................
...............

34

6.3

Genetically
-
engineered animal breeding colony/broodstock management

..............................

37

6.4

Archiving

................................
................................
................................
................................
......

38

6.5

Rederivation

................................
................................
................................
................................

39

7.

USE AND DISPOSITION OF GENETICALLY
-
ENGINEERED ANIMALS

................................
.................

41

7.1

Phenotyping and experimental procedures

................................
................................
................

41

7.2

Disposition/euthanasia

................................
................................
................................
...............

43

REFERENCES

................................
................................
................................
................................
....

45

GLOSSARY

................................
................................
................................
................................
.......

49

Appendix 1: Implementation of the Three Rs

................................
................................
....................

51

Appendix 2: Regulations

................................
................................
................................
...................

52

Appendix 3: Databases
for genetically
-
engineered animal lines

................................
.........................

54

Appendix 4: Welfare assessment

................................
................................
................................
......

55

Appendix 5: Nomenclature resources

................................
................................
...............................

62

Appendix 6: Example of a genetically
-
engineered animal passport

................................
....................

63

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1

1.

PREFACE

1. The Canadian Council on Animal Care (CCAC) is the national peer review agency responsible f
or
setting and maintaining standards for the ethical use and care of animals (vertebrates and
cephalopods) in science throughout Canada. The

CCAC publishes guidelines on
the general care and
use of animals

in science
,
and on issues of
current and emerging
ethical
concern. The
CCAC guidelines
on: genetically
-
engineered animals

used in science

rep
laces the
CCAC guidelines on: transgenic animals

(
CCAC, 1997
a)
. CCAC guidelines documents are published exclusively on the CCAC website,
which
should be consulted

for the most up
-
to
-
date versions.

2. CCAC guidelines are developed in response to advances in laboratory animal science,

emergence of
new technologies and the needs of the CCAC Assessment Program.

The
guidelines on genetically
-
engineered animals are provided to assist animal care committee (ACC) members and
researchers

in
evaluating the ethical aspects of the generation, ca
re, use and disposal of genetically
-
engineered
animals in accordance with the
CCAC policy statement on: ethics of animal investigation
(
CCA
C, 1989
)
.

3.
Review of the
CCAC guidelines on: transgenic animals
(
CCAC, 1997
a)

was prompted by the nee
d to
1) clarify the assignment of a category of invasiveness to protocols involving genetically
-
engineered
animals; and 2) review the implications of advancements in genetic engineering in relation to animal
welfare. The
CCAC guidelines on: transgenic anim
als

(
CCAC, 1997
a)

stated that new lines of genetically
-
engineered animals were to be classified as categor
y of invasiveness D (see
CCAC policy statement on:
categories of invasiveness in animal experiments

(
CCAC, 1991
)
), and re
-
clas
sified as appropriate when
the welfare of the animals was assessed. This reclassification was often not performed, leading to
misreporting of animal use in Canada. Additionally, during the development of the current guidelines
document the federal regulati
ons governing the use of genetically
-
engineered animals, the New
Substance Notification Regulations (Organisms), underwent review, and an attempt has been made to
align the current guidelines with the collection of information anticipated to be required by

the
Federal government, thereby reducing redundancy in reporting and regulatory burden on researchers.

4
.
It is recognized that many genetically
-
engineered animals do not have different welfare concerns
than conventional animals; however, the guidelines a
re aimed at timely identification of any welfare
concerns that may arise and the creation of appropriate strategies to address those concerns
. Other
issues associated with genetically
-
engineered animals that challenge implementation of the Three Rs
(
http://www.ccac.ca/en_/threer
)
include
the additional
animals
that
are used
in the
generat
ion of a

genetically
-
engineered animal

line

and invasive procedures used for generation, genotyping and
phenotyping.


5. These

guidelines are intended to provide assistance in the implementation of best practices and the
achievement of Russell and Burch’s Three Rs for use of animals in science
(
Russell and Burch, 1959
)
.
The CCAC recognizes that guidelines with the
term ‘should’ may

be subject to
interpretation by
properly constituted ACCs, and that in some case
s, an ACC may accept a different standard of practice
on the basis of adequate evidence and in keeping with the principles of the Three Rs. This discretion is
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August 2011


2

not extended to any other parties. Where the CCAC considers that no lower standard of practice is

acceptable, the term ‘must’ has been used. This document is intentionally broad to provide guidance
applicable to the range of genetically
-
engineered animals used in science; details relevant to particular
species will be

included in the guidelines docume
nts emerging from the revision of volume 2 of the
CCAC Guide to the Care and Use of Experimental Animals

(
CCAC, 1982
)
.

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3

SUMMARY OF THE GUIDELINES LISTED IN THIS
DOCUMENT

2.

G
eneral Considerations


Guideline 1:

All animals, whether genetically
-
engineered or not, are subject to the sa
me general principles that
govern the ethical treatment of animals, including the Three Rs.

Guideline 2:


A communication strategy should be in place to ensure that information relevant to the welfare of the
animals is effectively shared between research a
nd animal care staff.


Guideline 3:

P
rincipal investigator
s

must provide clear justification for the generation and/or use of a genetically
-
engineered animal.

Guideline 4:

Investigators must take every reasonable step to ensure they are not generating new
genetically
-
engineered animal lines when scientifically adequate lines exist and are accessible.

Guideline
5
:

A
nimal care committee
s that review protocols involving genetically
-
engineered animals

should include
persons with relevant expertise, and in the a
bsence of such expertise, should seek advice from
independent experts who can provide an understanding of the nature and likely implications

of the
proposed studies.

3. Facilities
,

Facility Management

and Staffing

Guideline 6:

Facilities housing geneticall
y
-
engineered animals must be designed in accordance with CCAC
guidelines.


Guideline 7:

Consideration should be given to any additional demands on staff time (animal care or research
team staff) which may be required for

the

welfare assessment of genetical
ly
-
engineered animals
and the implementation of appropriate mitigation strategies where welfare concerns are
identified.

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4

Guideline 8:

Investigators must ensure

their own competence, as well as that of
their staff
,

before generating
or using genetically
-
eng
ineered animals.

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5

4.

G
eneration and Welfare Assessment

Guideline 9:

Every reasonable effort must be made to ensure that the methods used to generate genetically
-
engineered animals are 1) appropriate for the species; 2) in line with the Three Rs; and 3) cap
able of
addressing the research objectives.

Guideline
10
:

Investigators must justify the methodology they will use to identify genetically
-
engineered founders
and
offspring
when requesting to generate a genetically
-
engineered animal line or acquire animals

for
which this is
unknown
.

Guideline 11:

The least invasive method of tissue collection and the minimum amount of tissue necessary to
ascertain the genotype should be used; re
-
sampling should be avoided. The use of more invasive
methods must be justified

and approved by the animal care committee.

Guideline 12:

Reasonably anticipated endpoints must be established before the generation of genetically
-
engineered animals and revised in the light of additional information on the effects of the genetic
modifica
tion on the animals’ welfare.

Guideline 13:

When investigators or core facilities produce new lines under protocols approved by the ACC, they
should report to the ACC on the efficiency of the methods used to genetically engineer the animals.


Guideline 14:

When investigators or core facilities generate genetically
-
engineered animal lines, procedures for the
generation and care of those animals must be part of an approved animal use protocol.


Guideline 15:

All genetically
-
engineered animal lines, including
those produced by crossing or backcrossing existing
lines, must undergo a welfare assessment. This assessment should be part of the documentation
accompanying the animals. A welfare status should be assigned to each line based on the welfare
assessment.

Gu
ideline 16:

If no welfare issues are identified, the new genetically
-
engineered animal line can be monitored to the
same extent as other animals with no welfare concerns. If welfare concerns are raised by an initial
welfare assessment, the principal invest
igator, veterinarian and ACC must be notified, and mitigation
strategies and/or appropriate endpoints
must

be developed in line with the research goals, which may
require a more extensive evaluation of the genetically
-
engineered animal line.

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6

Guideline 1
7
:

Veterinarians who report to an
animal care committee

should

assist in preparing standard operating
procedures for animal welfare assessment and establishment of appropriate endpoints for genetically
-
engineered animals.

Guideline
18
:

Effective monitoring a
nd mitigation strategies must be developed and used to minimize any pain or
distress observed or anticipated in genetically
-
engineered animals, unless required for the experiment
and approved by the animal care committee.

5.

Acquisition and Transportation

Guideline 19:

Genetically
-
engineered animals that are brought into or maintained in an institution should have
accompanying documentation, which should include the genotype or means of identifying genetically
-
engineered animals, along with information on
welfare concerns and mitigation strategies, if available.

Guideline

20:

Before genetically
-
engineered animals are brought into an institution, the protocol for their use and/or
further propagation must be approved by the
animal care committee
.

Guideline
21
:

As with the procurement of conventional animals used in science, investigators should ensure that
animals to be acquired have been generated in accordance with CCAC or equivalent standards.

Guideline
22
:

Genetically
-
engineered animals must be transported

in accordance with CCAC guidelines, with
consideration given to the impact of the genetic modification on the welfare of the animal
s

and
prevent
ing

release
of the animals
to the environment.

6.

Housing and Husbandry

Guideline
23
:

Genetically
-
engineered an
imals must be given correct standardized nomenclature for species where
such
standards

exist.

Guideline 24:

A genetically
-
engineered animal passport or similar documentation should be developed for each new
genetically
-
engineered animal line, be accessible

to the animal care committee, and accompany the
line if it is transferred to another institution.


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7

Guideline 25:

When completing the Animal Use Data Form for reporting annual animal use to the CCAC, genetically
-
engineered animals should be identified sepa
rately from non
-
genetically
-
engineered animals.


Guideline
26
:

The housing environment for genetically
-
engineered animals must be at least at the standard required
for conventional animals of the same species; however, the use of

strategies to address the
welfare
concerns of a genetically
-
engineered animal line may dictate the use of particular
housing and/
or
husbandry practices.

Guideline
27
:

Genetically
-
engineered animal breeding colonies/broodstock must be efficiently managed according to
approved protoc
ols, anticipated need, and the principle of reduction.

Guideline
28
:

Based on the Three Rs, investigators should use archiving technologies (e.g., cryopreservation) as a
reduction and sharing strategy for managing genetically
-
engineered animal breeding
col
onies/broodstock.

Guideline
29
:

As with all animals used in science, appropriate biosecurity measures to prevent genetic or pathogenic
contamination should be in place to minimize the need to rederive colonies/broodstock.

7.

Use and Disposition of Genetic
ally
-
Engineered Animals

Guideline
30
:

In assigning a category of invasiveness to an experimental protocol, as described in the
CCAC policy
statement on: categories of invasiveness in animal experiments

(
CCAC, 1991
)
, consideration must be
given to both the welfare status of the animals and the procedures to be performed.

Guideline
31
:

Any welfare concerns of the genetically
-
engine
ered animal must be taken into account when choosing
procedures.

Guideline 32:

Investigators should take reasonable steps to publicize all available phenotype and welfare
information, along with mitigation strategies.

Guideline
33
:

As with conventional a
nimals, care must be exercised in choosing the most appropriate time and
method of euthanasia, including for surplus animals produced by breeding colonies/broodstock.
Consideration must be given to the possibility that the phenotype of a genetically
-
engine
ered animal
could impact the effectiveness or humaneness of a given method

of euthanasia
.


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8


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9

2.

GENERAL CONSIDERATIONS

2.1

Definition of genetically
-
engineered animal and scope of this
guidelines document

6. For the purpose of this guidelines document, a
g
enetically
-
engineered animal

is a mammal,

bird,
fish, reptile, amphibian or cephalopod that has had a change in its nuclear or mitochondrial DNA (
e.g.,
addition, deletion or substitution of some part on the animal’s genetic material or insertion of foreign

DNA, including by chemical mutagenesis) achieved through a deliberate human technological
intervention. While this document focuses on the development and use of genetically
-
engineered
animal lines with germline transmission, animal welfare concerns apply

equally to those animals with
engineered transient expression of DNA, RNA and protein products that may affect the welfare of the
animals.

7
. This guidelines document covers
genetically
-
engineered founders as well as offspring

resulting from
natural breed
ing of genetically
-
engineered animals or genetically
-
engineered founders with
conventional

animals. Transgenic animals or transchromosomic animals that are engineered so as not
to transmit modifications to the next generation are also included within the s
cope of this document.

8
. The key feature of the CCAC definition of a genetically
-
engineered animal is that the genetic
modification
was

achieved deliberately through human technological intervention.

S
pontaneous
mutants that arise from time to time in bre
eding colonies and are then bred to develop a model of
human disease are not captured in the above definition
; however, the same welfare concerns arise
and analogous considerations apply
.

Similarly, marker
-
assisted breeding of livestock (i.e. conventional
selective breeding assisted by information on the genotype of the animal) can achieve an altered
phenotype, and can do so more rapidly than traditional selective breeding, but the modification is not
achieved through a technological intervention

at the gen
etic level
.

Although not included in these
guidelines, protocols involving spontaneous mutants and animals produced through marker
-
assisted
selection
(
Gibson and Bishop, 2005
)

require particular attention to address welfare concerns related to
alteration in
phenotype (see CCAC guidelines regarding the housing and husbandry of individual
speci
es) and the establishment of endpoints which minimize pain and distress
(see
CCAC guidelines on:
choosing an appropriate endpoint in experiments using animals for research

(
CCAC, 1998
)
).

9.
The previous version of th
is

guidelines
document
(
CCAC, 1997a
)

used the term
transgenic ani
mals
,
first introduced by Gordon
and

Ruddle

(
1981
)
. The term was broadly defined as “an animal in which
there has been a deliberate modificati
on of the genome


the material responsible for inherited
characteristics


in contrast to spontaneous mutation”

(
FELASA, 1995
)
.

10
.

The term
genetically
-
engineered animal

reflects more current terminology and captures the
broader spectrum of animals that can be generated through genetic engineering techniques.

All types
of animals

that fall under this definition are
covered by this
guidelines

document

for the following
reasons
: there is a possibility
that

expected or
unexpected effects on the phenotypes of these animals
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August 2011


10

could compromise animal welfare; the generation
, maintenance

a
nd use of these types of animals
raises ethical issues

and public
concern
(
e.g., Macnaghten, 2004
)
; and

these
types of animals
need to
be assessed for risk and housed according to federal, provincial and municipal regulations.

2.2

Ethical overview

Guideline 1:

All animals, whether
genetically
-
engineered or not, are subject to the same general principles that
govern the ethical treatment of animals, including the Three Rs.

1
1
. The CCAC's surveillance system for animals used in science is based on the principles of humane
science, i.e
. the Three Rs of Russell & Burch
(
1959
)



Reduction, Replacement and Refinement (see
http://www.ccac.ca/en_/threer

for a description of the Three Rs). These principles are laid out in the
CCAC policy statement on: ethics of animal investigation

(
CCAC, 1989
)
. The ethics of animal
investigation applies to all species covered by the CCAC system, i.e. all vertebrates and cephalopods,
including genetically
-
engineered animals.

12
.
The

generation and

use of genetically
-
engineered animals are subject to all of the considerations
raised by the
CCAC guidelines on: animal use protocol review

(
CCAC, 1997b
)
. Protocols involving
genetically
-
engineered animals
must

therefore be reviewed in the same manner as other protocols.
However, particular
attention
should

be

g
iven to the procedures involved and possible welfare
concerns for
genetically
-
engineered founders and
the
offspring

produced by breeding genetically
-
engineered animals (detailed in
s
ection
s

4.
2

Welfare
A
ssessment

and 7.1 Phenotyping and
Experimental Proced
ures
).

13
.

In implementing these guidelines, animal care committees
(ACCs)
and
investigators should take
into account the special features

of each genetically
-
engineered
animal
line
which may affect the
animals’
welfare
. In addition, they
should be sensiti
ve to ethical concerns and

alert to technological
changes in this rapidly evolving field

(see
Banner, 1998 and Appendix 1
)
.

14.
When reviewing a protocol,
ACCs

mus
t

weigh the potential harms the
genetic modification

and
associated procedures
may cause the research animals

and any adverse effects over the
experimental
lifetime of those animals (e.g., compromised welfare), against the potential benefits to humans, oth
er
animals or the environment that may result from the
studies
(
APC, 2003
;
CCAC, 1997
b
;
2000
)
. In this
context, it may be useful to consider the welfare o
f the genetically
-
engineered animals as
encompassing more than freedom from pain and distress. Definitions of animal welfare include
components of biological functioning and well
-
being, and recognize that there are both positive and
negative states of well
-
being
(
Fraser and Duncan, 1998
)
. The World Organisation for Anim
al Health
(OIE) defines animal welfare as “how an
animal

is coping with the conditions in which it lives” and
summarizes good animal welfare as follows:

An
animal

is in a good state of
welfare

if (as indicated by

scientific evidence) it is
h
ealthy, comfortable, well nourished, safe, able to express innate behaviour, and if it is
not suffering from unpleasant states such as pain, fear, and distress. Good
animal
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11

welfare

requires
disease

prevention and veterinary treatment, appropriate shelter,
management, nutrition, humane handling and humane
slaughter
/
killing
.


(
OIE, 2009; http://w
ww.oie.int/eng/normes/mcode/en_chapitre_1.7.1.htm
)

For genetically
-
engineered animals, this becomes important in ensuring that appropriate provisions
are made to mitigate any effects on animal welfare, anticipated or not, including establishing
appropria
te endpoints.

2.
2
.1

Applying the Three Rs

15
.

According to the
CCAC policy statement on: ethics of animal investigation

(
CCAC, 1989
)
,

it

is the
responsibility of the local
ACC

to ensure that animals are used only
after

the investigator's best efforts
to find a non
-
animal model have failed

(examples of information sources for
non
-
animal models

are
provided on the CCAC microsite,
http://www.ccac.ca/en/alternatives/index.html
)
. As for any ot
her
animals covered by the CCAC
, investigators using genetically
-
engineered animals are required to use
the most humane methods on the smallest
number of
appropriately selected
animals necessary to
obtain valid information.

Key concerns about the use of genetically
-
engineered animals include the
inherent inefficiency of the genetic engineering methods which generate surplus animals, the potential
pain and distress caused by
phenotypes with compromised welfare, unexpected welfare issues, and
issues particular to the husbandry and maintenance of breeding colonies. Responsibilities of
investigators to reduce animal use and refine their procedures are
described in Section 4.1, and
welfare assessment, monitoring and strategies to mitigate pain and distress are covered in
S
ection
s

4.2
and 4.3. Recommendations on housing and husbandry, good breeding colony/broodstock
management and archiving strategies are

provided in Section 5.

1
6
.

Consideration of how the tenet of the Three Rs may be applied to each study is essential to ensure
the humane use of genetically
-
engineered animals in
science
.

While the techniques for generating
genetically
-
engineered animals a
re continually changing, the guiding ethical
principles should always

be the Three Rs. As with the use of other animals, a balance is needed between using fewer animals
and repetitive
procedures on
the same animals
,

if the
procedures

result in substantial
pain being
inflicted on fewer animals. Examples of how the Three Rs might be implemented in the generation of
genetically
-
engineered animals are provided in


Appendix 1.

2.3

Communication

Guideline 2
:

A communication strategy should be in place to ensure t
hat information relevant to the welfare of
the animals is effectively shared between research and animal care staff.


17.
Because the
generation

and use of genetically
-
engineered animals require high levels of scientific,
technical and animal care expertis
e and skills, and because this work is typically shared between
research teams and animal care teams, communication of all relevant information between these
teams is crucial to the welfare of the animals, as well as to scientific success. Senior administr
ators
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12

must ensure that research and animal care teams can work constructively together, within an
appropriate institutional an
imal care and use structure (see the
CCAC policy statement for: senior
administrators responsible for animal care and use programs

(
CCAC, 2008
)
).


18.
When genetically
-
engineered animals are
generat
ed

in a core facility within the larger purpose
-
built, institutionally managed animal facilities, the
investigators

in the core facility should
be well
supported by
the ACC, veterinarian and animal care staff. Effective communication among these
groups is
essential to ensure protocols are properly followed and any animal welfare concerns are
documented and addressed.

19.
There should be a strong communications link between the ACC and the biosafety committee or
officer, such that protocols involving poten
tial biosecurity risks are properly assessed, with
documentation of the risks and requirements for mitigation measures (see Section 2.4.2
Responsibilities of ACCs).

2.4

Research
p
lanning

20
. The care and use of genetically
-
engineered animals fall within t
hree phases


generation, breeding
and phenotyping/experimental use


which are not necessarily distinct from each other. Animals
involved in the generation of a new genetically
-
engineered animal line and the breeding of subsequent
generations must be cove
red by an ACC
-
approved experimental protocol (see Section 4.1 Generation
of a new genetically
-
engineered animal line) until the welfare of the offspring has been assessed and
the ACC has determined the animals can be transferred to a breeding protocol (see

Section 6.2
Genetically
-
engineered animal breeding colony/broodstock management). Procedures to be
conducted to genotype and/or phenotype the animals must be included in the experimental protocol.
Additionally, any scientific use of those animals must be
covered by an approved experimental
protocol. All experimental protocols must be assigned a category of invasiveness to ensure the
appropriate level of care for the animals and proper reporting of animal use data to the CCAC (see
Section 7.1 Phenotyping an
d Experimental Procedures).

21.
All applicable federal, provincial/territorial and municipal regulations regarding the generation, use
and disposal of genetically
-
engineered animals must be followed. See Appendix 2 for a starting point
for consideration of

relevant regulations.

2
.
4
.1

Responsibilities of
Investigators

Guideline
3
:

P
rincipal
investigators must provide

clear justification for the generation and/or use

of a genetically
-
engineered animal.

22.
As with the use of other animals, investigators
must

only develop or use genetically
-
engineered
animals if there is no reasonable and practically accessible

alternative that will answer the research
question.

In line with

the Three Rs

tenet
, efforts
must

be made to explore other options for answering
the res
earch question.

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13

23
. The
investigator must justify to the ACC that using a scientifically validated research design that
involves a genetically
-
engineered animal line is the best way of answering the approved research
question in keeping with the CCAC requi
rements
.

In addition to the information required on an animal
use protocol form identified in the
CCAC guidelines on: animal use protocol review

(
CCAC, 1997
b)

and
the

CCAC terms of reference for animal care committees
(
CCAC, 2006
)
, investigators must include plans
for genotyping, welfare assessment (see Section 4.2 Welfare
a
ssessment), subsequent

on
-
going
monitoring of the welfare of the genetically
-
engineered animal

line and addressing potential welfare
concerns (see Section 4
.
3
Monitoring and mitigation strategies
). Once the protocol has received
approval from the ACC, then the particular
genetically
-
engineered animal
line may

be
brought into the
institution
,

or the

investigator
may
generate the animal
s

or undertake to have the animal
s

generated
by a service provider.

Guideline 4:

Investigators must take every reasonable step to ensure they are not generating new genetically
-
engineered animal lines when scientificall
y adequate lines exist and are accessible.

24
. There are many confounding issues that may prevent the use of a pre
-
existing animal model. These
include intellectual property protection; scientific barriers; quality of the animals; incorrect analysis or
use

of a compromised background strain; health status; availability; and

accessibility. However,
investigators should perform

a literature search and consult the available databases to ensure their
research goals cannot be met with an already existing, availa
ble and accessible genetically
-
engineered
animal line, and should be able to provide evidence that this has been done (see Appendix 3 for a list
of potential databases).

2
5
. The protocol for the generation of a new line should include the
elements identif
ied in the
CCAC
guidelines on: animal use protocol review

(
CCAC, 1997
b)
,
as well as the
following:



a form to be used to report the general condition of the animals generated, which includes
anticipated effects of the genetic modification and the basis for these predictions (see Appendix
4

for an example);



the identification of

an SOP for monitoring the animals during their experimental lifetime; and



a statement regarding the expected length of time required until the welfare status of the line can
be determined (see Section 4.2 Welfare
a
ssessment).

2.
4
.2

Responsibilities
of
A
CC
s


Guideline
5
:

Animal care committees that review protocols involving genetically
-
engineered animals

should
include persons with relevant expertise, and in the absence of such expertise, should seek advice
from independent experts who can provide an und
erstanding of the nature and likely implications of
the proposed studies.

26
. The
CCAC terms of reference for animal care committees

(
CCAC, 2006
)

should be consulted for
information on the general roles and responsibilities of institutional ACCs.

Draft


August 2011


14

27. ACCs should ensure that sufficient expertise is available to provide a review of protocols
conce
rning the generation and use of genetically
-
engineered animals. This

may require providing
training for ACC members, in particular to assist in the understanding of commonly used terms and
methods.

28
. ACCs that regularly deal with projects involving genet
ically
-
engineered animals should have two or
more professionals with experience in this area on the committee. Committees that rarely review
studies involving genetically
-
engineered animals may need to rely on
ad hoc

expert advice regarding
the potential w
elfare implications of proposed techniques on the individual animals. However, given
the wide range of species and methods employed, and the rapid evolution of technology, even
committees highly experienced with reviewing protocols involving genetically
-
en
gineered animals may
need to periodically seek outside reviews from other professionals.

29
. The ACC should be attentive to matters such as

the type of anesthetic and analgesic to be used; the
welfare assessment method that will be used (recording sheets
and other supporting documents
should be provided by the investigator); endpoints; euthanasia method; disposition of “surplus”
animals; the management of breeding colonies, including screening protocols to identify appropriate
animals; and characteristics
of the housing and husbandry for the animals (e.g., barrier vs.
conventional facilities).

3
0
. The ACC should work with the investigator and animal care staff to assign an appropriate category
of invasiveness in animal experiments and develop a list of inst
ructions for monitoring the animals and
dealing with any adverse effects (intended or unanticipated) resulting from the genetic modification
(
s
ee
S
ections 4.2 Welfare
a
ssessment, 4.3 Monitoring and
m
itigation
s
trategies and 7.1 Phenotyping
and
e
xperimental

p
rocedures). The breeding strategy may depend on the impact of the modification
on the phenotype and/or fertility of the animal (see Section 6.2 Genetically
-
engineered animal
breeding colony/broodstock management).

31
. A
system must be

established to ensu
re that where the methods used to generate the genetically
-
engineered animals or the animals themselves have potential biosafety concerns, the protocols are
forwarded to the biosafety committee/officer for review prior to approval by the ACC. The biosafety

committee or officer must ensure the organism is appropriately classified and adequate risk
assessment is undertaken to be able to ascertain the necessary conditions for housing and husbandry
of the animals and their subsequent disposal.

2.4.3

Responsibil
ities of Veterinarians

32
. Veterinarians who report to an ACC must have input into the standard operating procedures (SOPs)
for the housing and husbandry of the animals, animal welfare assessment, and the establishment and
recognition of appropriate endpoi
nts. The veterinarian bears the final responsibility for
decisions
regarding
euthanasia of an animal if the endpoint has been reached or if an animal experiences
unrelievable pain and/or distress that has not been approved by the ACC as part of

a

research
protocol
(
CCAC, 1998
)
.

Draft


August 2011


15

33
. Veterinarians working in facilities where genetically
-
engineered animal lines are being generated
and used should have

adequate
knowledge to understand the terminology, generati
on techniques,
screening procedures, and phenotyping techniques, and be able to provide appropriate care for the
species involved. In some cases, it may be necessary to consult other veterinarians with experience in
the welfare implications of proposed tec
hniques on particular animals. The veterinarians should be
familiar with any animal
-
based procedures to be used by investigators in their institution, and provide
advice in developing protocols in line with the Three Rs. The veterinarians and senior animal

care staff
should also provide appropriate training to others involved in the care of genetically
-
engineered

animals when needed.


Draft


August 2011


16

3.

FACILITIES, FACILITY
MANAGEMENT AND STAFFING

3.1

Facilities and
f
acility
m
anagement

Guideline 6:

Facilities housing gene
tically
-
engineered animals must be designed in accordance with CCAC
guidelines.


34
. The

basic

requirements for facilities that house genetically
-
engineered animals are the same as for
conventional animals,
as detailed in

the
CCAC guidelines on: laboratory

animal facilities

-

characteristics, design, and development

(
CCAC, 2003a
)
.

However, some asp
ects of facility design and
maintenance are particularly important for

some

genetically
-
engineered animals

and the facility
manager and investigator need to discuss the requirements of the animals to make sure that the
facility can provide for their needs.

Additionally, specific
SOPs must be in place to prevent the release
of
genetically
-
engineered
animals into the environment.

35
. Genetically
-
engineered animals may require specialized equipment
for phenotypic analysis
(e.g., equipment for imaging, cardiov
ascular monitoring, etc.)
.
Properly equipped procedure
rooms
within purpose
-
built animal facilities
are important to eliminate the need to transport the
animals, and thus ensure the health of the animals is not compromised, reduce the
introduction of exper
imental variables, and maintain the necessary conditions for
confinement of the
genetically
-
engineered
animals.

When this equipment
is not available in
local procedure rooms,

appropriate transport SOPs must be in place (as noted in Section 5.2
Transportati
on).

3.1.1

Responsibilities of Institutions

36. There must be competent veterinary and animal care service providers whose numbers and
expertise match the nature and scope of the institutional program
, as noted in the
CCAC policy
statement for: senior admi
nistrators responsible f
or animal care and use programs

(
CCAC, 2008
)
.

37. A senior institutional official must be identified in the Terms of Reference of the institution’s ACC
as having overall responsibility for the institution’s animal care and use program. This person must
ensure that appropriate animal care and use serv
ices are provided to meet the institution’s scientific
goals, and that all animal care and use is conducted according to institutional and CCAC policies and
guidelines
(
CCAC, 2008
)
.

3.1.2

Responsibilities of ACCs

38
. ACCs should work with facility managers to ensure that appropriate facilities and equipment are
available, e
ither at the home institution or through a pre
-
arranged agreement with another institution,
prior to the approval of protocols involving the introduction or generation of new genetically
-
engineered animals.

Draft


August 2011


17

3.1.3

Responsibilities of Facility Managers

39
.
Facility managers are responsible for the day
-
to
-
day management of the animal care facilities and
the daily care of the animals housed there
in
.

40
.

It is the responsibility of the facility manage
r

to work with the
ACC

to
determine the animal facility
capac
ity required to house the genetically
-
engineered animals.
Large numbers of animals
may be

required in the generation of genetically
-
engineered animals

and

the subsequent maintenance of
breeding colonies
/broodstock.

F
acilit
y
managers must therefore

ensure t
hat sufficient resources are
available to house and care for the anticipated

numbers of animals involved.

41
. Facility managers play a key role in encouraging reduction and refinement, such as through
implementing SOPs for efficient breeding colony/broodst
ock management, and for housing and
husbandry strategies that seek to optimize the welfare of animals in the research setting. They must
work with the veterinarian and ACC to ensure that SOPs for health surveillance and containment of
genetically
-
engineere
d animals are developed and implemented.

42
. Genetically
-
engineered animals must be kept under confined or contained conditions in
accordance with government regulations and must not be released to the environment (see Appendix
2
). Strict procedures to pre
vent the incidence of zoonotic disease transmission
must

be in place,
especially when the animal is generated for a challenge or infectivity related experiment (immune
compromised). Routine and specific sanitation measures should be employed according to t
he
assessment of the veterinarian supervising the genetically
-
engineered animals. Where necessary,
medical
advice should be sought

concerning risks for transmission of zoonoses.

3.1.4

Responsibilities of Investigators

43
. Investigators must collaborate wit
h the ACC and authorities within the institution to ensure that
appropriate information concerning their research protocol(s) is submitted to the relevant regulatory
agencies. Investigators must comply with federal, provincial and local regulations regardi
ng any
environmental effects, disposal and maintenance of genetically
-
engineered animals. Appendix 2
provides a starting point for consideration of relevant regulations.


3.2

Staffing

44.
As noted in the Preface, many genetically
-
engineered animals do not
have different welfare
concerns than conventional animals; however, the guidelines are aimed at timely identification of any
welfare concerns that may arise and the creation of appropriate strategies to address those concerns.

Guideline 7:

Consideration sh
ould be given to any additional demands on staff time (animal care or research
team staff) which may be required for
the
welfare assessment of genetically
-
engineered animals and
the implementation of appropriate mitigation strategies where welfare concerns

are identified.

45
. As described in
S
ection 4.2 Welfare
a
ssessment, all new genetically
-
engineered animal lines must
undergo a welfare assessment. If no welfare issues are identified, the new line can be monitored to
Draft


August 2011


18

the same extent as other animals with
no welfare concerns. However, if welfare concerns are
identified, a more extensive evaluation of the line should be carried out to develop appropriate
mitigation strategies. Therefore, maintaining genetically
-
engineered animals may require investing
more t
ime and resources into husbandry and observation whe
n

there are animal welfare concerns.
This includes the time and training necessary to carry out
in depth
welfare assessment of the
animals.
Genetically
-
engineered animals should not be generated or brough
t into a facility unless there are
sufficient human resources to perform the welfare assessment and apply appropriate mitigation
strategies to address anticipated welfare concerns.

3.3.1

Responsibilities of Institutions

46
. The institution is responsible f
or ensuring that staff is trained for the facility maintenance and
animal care tasks required by their position, and are aware of the associated risks and how to minimize
them.

3.3.2

Responsibilities of
I
nvestigators

Guideline 8:

Investigators must ensure
the
ir own competence, as well as that of their
staff
,
before generating or
using genetically
-
engineered animals.

47
. Investigators, graduate students and research assistants/technicians should complete appropriate
theoretical training as outlined in the
Re
commended Syllabus for an Institutional Animal User Training
Program

(
CCAC, 1999
)

and all those who will handle animals need to have the necessary technical skills
which are acquired through practical training. This training should be ke
pt up
-
to
-
date, as best practices
will continue to evolve.

Draft


August 2011


19

4.

GENERATION AND WELFARE ASSESSMENT

4.1

Generation of a new genetically
-
engineered animal line

48. Some methods of genetic engineering are more efficient in terms of reduction and refinement of
an
imal use in some species than in others, and not all methods are currently available for use in all
species. Methods such as targeted mutagenesis, targeted gene replacement, and conditional or
inducible gene constructs
,
which allow control of when and wher
e a gene of interest will be expressed
or silenced, may help prevent unintended negative effects on the welfare of genetically
-
engineered
animals.

49. In institutions where a number of genetically
-
engineered animal lines are developed, core facilities
may

be available that include the necessary expertise and equipment. Use of these facilities can help
ensure effective methods of producing animals are used, and may reduce numbers of animals
involved. As noted in Section 2.3 Communication, it is important th
at there is effective communication
among scientific personnel in the core facilities, veterinary and animal care staff, and the ACC to
ensure that the Three Rs are well applied; institutional SOPs, particularly those on anesthesia,
analgesia and surgical
procedures, are respected; and SOPs specific to core facilities are developed as
necessary.

50. As soon as the genetically
-
engineered animal with the desired genetic modification or phenotypic
trait is identified, a breeding strategy is needed to further
propagate the animals. The genetically
-
engineered founders and their offspring require welfare assessment (see Section 4.2 Welfare
assessment) to identify and document any welfare concerns and determine the need for additional
monitoring and/or mitigation
strategies (see Section 4.3 Monitoring and mitigation strategies).

51. The breeding procedure should be regarded as an experiment at least until a welfare assessment
has been performed and information regarding welfare concerns and mitigation strategies ha
s been
documented by the investigator and forwarded to the ACC. During this phase, a category of
invasiveness should be assigned to the protocol, as explained in Section 7.1 Phenotyping and
experimental procedures. The investigator should report back to th
e ACC as soon as feasible on any
additional information related to animal welfare that is obtained through further phenotyping.

52. Animals that have been used for non
-
germline genetic manipulation should not normally be bred.
Where the research goals requ
ire these animals to be bred, special approval must be obtained from an
ACC, giving appropriate consideration to the impact of the manipulation on the welfare of the animals.

53. Stable germline transmission does not necessarily equate to a stable phenoty
pe and animal
welfare, since phenotypes can change (e.g., age
-
dependent phenotypes, background effects,
homozygosity, and breeding to other mutant lines). Therefore, appropriate monitoring is needed
whenever the experimental lifespan of an animal is extend
ed or the genetic background or gene
-
specific genotype is changed.

Draft


August 2011


20

4.1.1

Responsibilities of Investigators

Guideline 9:

Every reasonable effort must be made to ensure that the methods used to generate genetically
-
engineered animals are 1) appropriate for

the species; 2) in line with the Three Rs; and 3) capable of
addressing the research objectives.

54.
To ensure efficient generation of genetically
-
engineered animals,
investigators should

make
certain
that staff are well

trained and have demonstrated prof
iciency in performing the
techniques required
and in humane handling of the animals. Where facilities
with expertise in
genetic engineering

of
animal
s

exist and work according to appropriate, ACC
-
approved procedures
,
they should be used to
generate new gen
etically
-
engineered animals or their expertise should be sought. Investigators must
provide scientific justification if they do not intend to use reputable core facilities; cost should

not be
the primary justification
.

55
.

Investigators must

adequately jus
tify the genetic manipulation introduced

in view of the
scientific
questions
being
ad
d
ress
ed
.
Investigators should attempt

to refine their generation protocols and
incorporate best practices to minimize the numbers of animals involved in generation of an a
nimal
model and minimize any potential pain and distress.

56.

Crossing existing genetically
-
engineered animal
lines (e.g., to create compound mutants)

can have
phenotypic effects
, anticipated or not, that affect animal welfare,

and is considered to be
the
generation of a new line; therefore, the same protocol review outlined in Section 2.4 Research
p
lanning applies.

Guideline
10
:

Investigators must justify the methodology they will use to identify genetically
-
engineered founders
and
offspring

when requesti
ng to generate a genetically
-
engineered animal line or acquire animals
for which this is unknown.

5
7. Rapid and accurate identification of genetically
-
engineered founders and
offspring

is a necessary
pre
-
requisite for compliance with the Three Rs in the us
e of genetically
-
engineered animals.

Guideline 11:

The least invasive method of tissue collection and the minimum amount of tissue necessary to
ascertain the genotype should be used; re
-
sampling should be avoided. The use of more invasive
methods must be

justified and approved by the animal care committee.

5
8. With larger animals (e.g., cattle and pigs), it is relatively easy to use less invasive procedures for the
collection of tissue samples for genotyping. However, for smaller species (e.g., mice and z
ebrafish),
more invasive methods may be required to obtain sufficient tissue for DNA extraction; in this case,
relevant anesthesia/analgesia regimes should be examined by the ACC and applied as necessary.
Tissue obtained at the time of ear punching (mammal
s) or fin clipping (fish) for identification purposes
should be used for genotyping, rather than using additional
tissue biopsy
procedures.

Draft


August 2011


21

5
9. The techniques chosen should provide sufficient material to ensure that repeat sampling is not
required. If two

samples are required, the sampling interval should be specified and

approved by the
ACC. Re
-
sampling should be

avoided unless identified as a necessary requirement of the experimental
protocol.

Guideline 12:

Reasonably anticipated endpoints must be establ
ished before the generation of genetically
-
engineered animals and revised in the light of additional information on the effects of the genetic
modification on the animals’ welfare.

60
. Reasonably anticipated welfare concerns
must

be considered and endpoint
s established in
consultation with the veterinarian and the ACC before the generation of genetically
-
engineered
animals, recognizing that some phenotypic effects may not be known in advance (see the
CCAC
guidelines on: choosing an appropriate endpoint in e
xperiments using animals for research, teaching
and testing

(
CCAC, 1998
)
). Pilot projects should be considered in establishing endpoints when there is
insufficient information from which to make reasonable

predictions. The endpoints should then be
revised in light of the welfare assessment (see Section 4.2 Welfare
a
ssessment) and observed
phenotypic characteristics, particularly
in cases

with a negative impact on the animals’ welfare.

61
. The agreed endpoi
nts should become part of the animal passport accompanying the animal line
(see Section 6.1
I
dentification and
r
ecord
k
eeping), so that ACCs in institutions where the line may be
used have the information available when considering protocols involving the
use of these animals.


Guideline 13:

When investigators or core facilities produce new lines under protocols approved by the ACC, they
should report to the ACC on the efficiency of the methods used to genetically engineer the animals.


62
. Prior to produc
ing a new line, investigators should determine appropriate benchmarks for
successfully producing the new line in accordance with the Three Rs, based on
available scientific
literature and
in
consultation with others having relevant expertise. Deviation fro
m these should be
justified to the ACC, and remedial action should be taken to correct problems if the success rate is low
in relation to the current standards in the field.

63
. When investigators or core facilities produce new animal lines, they should k
eep records of the
efficiency of the generation program. These records should include the following information noted by
Wolfensohn and Lloyd
(
2003
)
:



the number of animals used per DNA construct

or ES cell line

(keeping in mind the different
categories of animals, such as embryo sources, breeding males, su
rrogate mothers, etc. in the case
of mammals, and gene transferred eggs in the case of fish or amphibians);



pregnancy/fertility rates;



percentage success in generating viable new modifications (where this is not known until a later
life stage, estimates
should be made and confirmed when results are obtained);

Draft


August 2011


22



the genotypes/phenotypes produced (e.g., mosaics, chimeras, non
-
takes (i.e. “wild
-
type” animals
that do not carry the
transgene), non
-
chimeras (i.e. wild
-
type that do not carry the ES cell line)
an
d unwanted genotypes)
;



the results of health screening and any deaths or welfare problems identified; and



the breeding performance of each genetically
-
engineered
animal
line and the fate of the animals
produced
.



64.
This
information
c
an be used to

iden
tify inefficiencies in production of genetically
-
engineered
animals, and lead to re
-
dressing these inefficiencies to reduce the number of animals used.


65. Where an animal line has been produced by an investigator or core facility, the efficiency of the
g
enetic engineering program in general should be reported to the ACC as part of the generation
protocol and at the protocol renewal stage.


4.
1
.2

Responsibilities of ACCs

Guideline 14:

When investigators or core facilities generate genetically
-
engineered a
nimal lines, p
rocedures for
the generation and care of
those
animals must be

part of an approved animal use
protocol
.

66
. Standard Operating Procedures

(SOPs)

for

the generation of genetically
-
engineered
animals should
be reviewed by the ACC
as needed, i
n
keeping with the rapidly evolving nature of the field of genetic
engineering and with continual advances in research on animal welfare. SOP
s

describing the specific
procedures to be used and the age of the animal
(s)

involved in the procedure should be deve
loped in
accordance with the Three Rs and approved by the veterinarian and ACC. ACCs must be sure to also
evaluate the impact of the restraint on the animal when approving protocol
s
.

67
. Core facilities should be subject to external review on a regular bas
is to ensure they are in keeping
with best practices. Where centralized core facilities provide services to produce genetically
-
engineered animals, external scientific and procedural reviews may be appropriate to identify and
assist with implementation of
best practices.

4.1.3

Responsibilities of Veterinarians

68
. The veterinarians should be familiar with any special care needed to handle genetically
-
engineered
animals, breeding colonies and
neonates, and should be aware of any potential complications that
may result from the alteration to the animals’ genome.

69
. The veterinarian should assist in the development of SOPs for the generation of genetically
-
engineered animals and

their monitoring, and in the establishment of endpoints.

4.2

Welfare assessment


7
0
. The effects of genetic engineering on animal welfare is of significant public concern
(
Mench, 1999
)

and has been documented for a number of species (for example, see NRC
(
2002
)
; Hallerman
et al
.
(
2007
)

and Wells et al.
(
2006
)
)
. These publications and others point to

the need to
perform
good
Draft


August 2011


23

welfare
assessment of genetically
-
engineered animals, not only on founder animals but also on
subsequent generations. They also indicate that common husbandry
practices (e.g.
,

backcrossing)

may
have unanticipated consequences for genetically
-
engineered animals.


71. Careful welfare assessment can help staff to determine which lines may have compromised
welfare. In turn, information obtained during characterization of the pheno
type of a line can assist
with identification of parameters which might be useful in devising appropriate welfare assessment
SOPs. Where possible, controlled language for observation will assist in the clear understanding of
welfare issues (see
http://mousewelfareterms.org
).

72.
Additional
human

resources
may be

needed to carry out welfare assessment

(as noted in Section
3.3 Staffing)
.
Welfare assessment often involves the investigator, veterinarian, research assis
tants and
animal care staff, and requires an efficient system of reporting and communication (see Section 2.3
Communication).

4.2.1

Responsibilities of Investigators

Guideline 15:

All genetically
-
engineered animal lines, including those produced by crossi
ng or backcrossing
existing lines, must undergo a welfare assessment. This assessment should be part of the
documentation accompanying the animals. A welfare status should be assigned to each line based
on the welfare assessment.

73. Welfare assessments ar
e intended to identify welfare concerns that arise as a result of the genetic
modification, both anticipated and unexpected. The investigator should ensure that those who will be
conducting the welfare assessment have been identified in advance (e.g., them
selves, trainees
research assistants/technicians or animal care staff), and that they are competent in performing the
required assessment and recording the information. This may involve consultation with the
veterinarian and facility manager.

74. Investiga
tors must properly document the welfare assessment and identification, if applicable, of
animal welfare
-
related issues of newly generated genetically
-
engineered animal lines
,

along with any

mitigation strategies, and report these back to the
ACC
. The welfa
re assessment can be done in
parallel with research on new genetically
-
engineered
animal lines, and any additional welfare
information that is acquired through the research should be communicated to others involved in the
assessment and care of the animals

(see Section 2.3 Communication).

75. The welfare assessment should encompass the normal experimental lifespan of the animal,
starting at birth
,

and be initiated as soon as possible after generation (for examples, see Appendix 4
and the R
eport of the BVAAW
F/FRAME/ RSPCA/UFAW Joint Working Group on Refinement
-

Defining and implementing protocols for the welfare assessment of laborato
ry animals
(
Hawkins et al., 2011
)
). The assessment and intervals at which the animals are to be assessed should
be designed by the investigator in consultation with exper
ts on the particular species and the
veterinarian(s). It should be comprehensive and rigorous enough so that the welfare of the line is
Draft


August 2011


24

effectively assessed, keeping in mind the expected phenotype. The welfare assessment form should be
treated as an SOP an
d approved by the ACC. Even if it does not appear that there will be welfare
issues, an assessment of subsequent generations should be performed, as welfare problems may not
emerge in the first few generations, particularly when genetically
-
engineered anim
als are made
homozygous by subsequent generations of breeding.

76
. When genetically
-
engineered animal lines are acquired from external suppliers (see Section 5.1
Acquisition of existing genetically
-
engineered animal lines) and the accompanying

animal welf
are
information is determined to be sufficient to assign a welfare status, as defined below,

a
n initial

welfare assessment is not required. The
information should be recorded in the in
-
house system and
made accessible to authorized personnel. Imported line
s which have not been previously assessed, or
for which welfare
-
related information is lacking, should be

subjected to the appropriate level of
assessment
.

77.
All genetically
-
engineered animal
lines should be

assigned a welfare status based on the welfare

assessment. Welfare status provides a general indication of the level of monitoring and mitigation
required for the animals lines, and is defined as follows:







78.
The welfare status of the animals needs to be taken into consideration in assigning a
category of
invasiveness to protocols involving the use of those animals (see Section 7.1 Phenotyping and
e
xperimental
p
rocedures), as experimental procedures may have a greater impact on animals with
compromised welfare. New genetically
-
engineered animal
lines
should

be assigned a welfare status of
‘pending’ until their welfare has been assessed.

79. Figure 1 provides a flow chart of the recommendations for assessment of the welfare of
genetically
-
engineered animals.

Guideline 16:

If no welfare issues are
identified, the new genetically
-
engineered animal line can be monitored to
the same extent as other animals with no welfare concerns. If welfare concerns are raised by an
initial welfare assessment, the principal investigator, veterinarian and ACC must be
notified, and
mitigation strategies and/or appropriate endpoints must be developed in line with the research
goals, which may require a more extensive evaluation of the genetically
-
engineered animal line.

80.
Where there are animal welfare concerns, the
li
ne should be

evaluated
beyond
the initial

assessment using appropriate tests or observations
.

Information relevant to the welfare of the animals


Standard


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Mild to moderate


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be敮e iT敮瑩晩敤Ⱐ 慬ong 睩瑨t慰p牯p物r瑥
m楴楧慴aon⁳ r慴ag楥s



Severe


睥汦l牥r con捥cn猠 U慶攠 b敥n 楤敮瑩晩敤e 瑨慴t 牥煵楲攠 數瑥n獩v攠 m楴楧慴楯n
m敡獵e敳e慮T⁣lo獥 mon楴o物rg

Draft


August 2011


25

can additionally be obtained from the results of phenotyping screens. Any relevant information
obtained from ph
enotyping must be shared with the ACC and the animal care staff to assist in
providing appropriate care for the animals. The
information should also be

added to an
animal
passport

(see Section 6.1 Identification and
r
ecord
-
keeping) and made available to ot
her potential
users of those animals through publication or an online

database
.

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August 2011


26

Figure 1

Assessment of the Welfare of Genetically
-
engineered
Animals

(
Ada
pted from Wells et al., 2006
)
























Animals Acquired or Generated

New genetically
-
engineered animal line is acquired
or generated.

Conduct a Welfare Assessment

Carry out a welfare assessment according to an ACC
-
approved SOP, using a list of key indicators of welfare.
Record results and numbers of animals assessed.

Is a welfare
concern
identified?

NO

YES

Record Conc
erns & Determine Appropriate Action

Make a full record of the signs observed and inform the
veterinarian, principal investigator and ACC.

Appropriate action is applied as determined by the ACC
working with the veterinarian and principal investigator (e.g.
,
more extensive welfare assessment, more frequent monitoring,
mitigation measures, or appropriate

endpoints
)
.



If a welfare
concern is

identified.

Continue Assessment & Monitoring

Continue welfare assessment, monitoring and recording as
determined by t
he ACC, with the application of approved
mitigation strategies and/or endpoints. Evaluate any additional
information available from phenotyping that might impact animal
welfare and record any additional welfare concerns.


Protocol Approval

Animal use pro
tocol for
the generation and/or use of a genetically
-
engineered animal line
and its handling is prepared by the investigator and approved by the ACC.

Assign Welfare Status & Monitor

Assign status of ‘standard’ and
monitor at the level of conventional
anim
als without welfare concerns.
Evaluate any additional information
available from phenotyping that
might impact animal welfare.


Assign Welfare Status

Assign a status of ‘mild to moderate’ or ‘severe’,
depending on the impact of the welfare concern on the
animals and the availability of mitigation measures.

Is sufficient information
on the welfare of the
animals available?

NO

YES

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27



Draft


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28

81.
Progress in the welfare assessment and/or the welfare
record sheets

and mitigation
strategies (see
Section 4.3 Monitoring and mitigation
strategies) must be reported to the ACC in a timely mann
er,
minimally at each annual protocol renewal. Therefore, progress on welfare assessment must be
reported no later than one year afte
r the onset of the generation protocol.
F
or longer
-
lived species
with a longer maturation period, it may take several proto
col renewal cycles before sufficient
information can be gained about the phenotype of the animals and its effect on welfare, particularly
for traits that appear later in the lifecycle. Nevertheless, protocol renewals
must

include an update on
progress towa
rd the generation
and welfare

assessment of the genetically
-
engineered
animal line
throughout its experimental

life cycle
. Even with mice, rats and zebrafish, which have relatively short
lifecycles and rapid maturation, some later
-
onset traits may not be d
etected within the first one
-
year
period, especially since it takes several months or breeding cycles before the first generation of
homozygous animals is produced. Homozygous,

heterozygous

and hemizygous
(in the case of
transgenics)

animals should all be
monitored for phenotypic effects of the genetic modification that
affect their welfare, and these should be reported. Animals found to be experiencing unrelievable pain
and/or distress

not approved as part of the research
protocol should be

euthanized.

4.2
.2

Responsibilities of ACCs

82. Procedures
for animal welfare assessment must be reviewed and approved by the ACC. The
y

should indicate

the number of animals to be assessed, the key indicators that will be used and the
appropriate life stage for assessmen
t of the animals. Where possible,
a

basic SOP that has been
approved by the ACC should be used and adapted to the specific animal line

to improve efficiency
.

4.
2
.3

Responsibilities

of Veterinarian
s

Guideline 1
7
:

Veterinarians who report to an animal care
c
ommittee

should

assist

in preparing standard operating
procedures for animal welfare assessment and establishment of appropriate endpoints for
genetically
-
engineered animals.


83.
The
veterinarian should assist investigators in the development of welfare a
ssessment procedures
for lines being produced or acquired.

84. When welfare concerns are identified, the veterinarian should work with the investigator to
establish the appropriate course of action.

85
.
Veterinarians and facility managers

are in a positio
n to pass on acquired knowledge on the welfare
implications of practices involving genetically
-
engineered
animals and should

document this acquired
knowledge as a reference for subsequent reviews.

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29

4
.
3

Monitoring and mitigation strategies

Guideline
18
:

Effe
ctive monitoring and mitigation strategies
must b
e developed and used to minimize any pain or
distress
observed or anticipated
in genetically
-
engineered animals, unless required for the
experiment and approved by the animal care committee.

86
. Information
gained from the welfare assessment
process should be used to develop genetically
-
engineered animal welfare record sheets for monitoring the animals and to devise mitigation
strategies, if needed. Once the record sheets and mitigation strategies have been e
valuated for
effectiveness by the investigator or designate and veterinarian, this information should be added to a
genetically
-
engineered animal passport (see Section 6.1 Identification and
r
ecord
-
keeping).

87. Regular monitoring of behavioural and other
characteristics of animals that have been subject to
genetic engineering and subsequent generations should be documented, preferably electronically.
Record sheets, or an equivalent method, should be tailored to the

genetically
-
engineered
animal line

and ag
e of the animals.


88
. Adverse effects on the welfare of genetically
-
engineered animals can be mitigated in some cases.
For example, some genetically
-
engineered mice with compromised locomotion benefit from providing
food on the cage floor, rather than in
the hopper alone. Analgesics can help with chronic pain. For all
species, genetically
-
engineered animals with phenotypes giving rise to compromised
welfare should be
identified as quickly as possible so that appropriate mitigation strategies and endpoints
can be
implemented
(
Robinson et al., 2003
)
.

4.3.1

Responsibilities of Investigators

89. I
nvestigators should

ensure that an ACC
-
approved
procedure

for monitoring the animals during
their experimental lifetime is in place, and that all relevant welfare
information

gathered through
welfare assessment, monitoring or phenotyping is properly docume
nted.

4.3.
2

Responsibilities of ACCs

90
. ACCs must review and approve procedures for monitoring genetically
-
engineered animals.

4.3.3

Responsibilities of Facility Managers


91. The facility manager must ensure that approved procedures for monitoring animal
s are followed
and that information relevant

to the welfare of the animals is properly recorded and communicated to
the investigator and veterinarian.

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30

5.

ACQUISITION

AND
TRANSPORTATION

5
.1

Acquisition of existing genetically
-
engineered animal lines

Guide
line 1
9
:

Genetically
-
engineered animals that are brought into or maintained in an institution should have
accompanying documentation, which should include the genotype or means of identifying
genetically
-
engineered animals, along with information on welfar
e concerns and mitigation
strategies, if available.

92
. As noted in the
CCAC guidelines on: procurement of animals used in science

(
CCAC, 2007
)
, Section
6.1 Documentation, in addition to the information mentioned above, the ACC should also be provided
with the following:



the genetic background (inbred

or outbred
,

if applicable
)
;




name (common and
standardized
nomenclature)

and
reference if available
; and




health status and any special care required
.

This information should be in a passport for the genetically
-
engineered animal line (see Section 6.1
Identification and
r
ecord
-
keeping), which is sent ahead electronically. A paper
copy of the passport
should accompany the animals.


93.
If the provider of the genetically
-
engineered line cannot supply the necessary information, it
becomes the investigator’s responsibility to generate the information and provide it to the
ACC.
Acquisit
ion of genetically
-
engineered animals without documentation may be considered under
circumstances where the acquired genetically
-
engineered animals will be treated as a newly generated
line and the data for the passport is accumulated and transmitted to th
e ACC after import of the
animals.


5.1.1

Responsibilities of Investigators

Guideline

20:

Before genetically
-
engineered animals are brought into an institution, the protocol for their use
and/or further propagation must be approved by the ACC.

Guideline
21
:

As with the procurement of conventional animals used in science, investigators should ensure that
animals to be acquired have been generated in accordance with CCAC or equivalent standards.

94.
Guidelines for acquisition of any animals used in science ar
e set out in the
CCAC guidelines on:
procurement of animals used in science

(
CCAC, 2007
)
. Section 4. Sources of Animals stat
es that
“animals should be obtained from reputable suppliers” and provides further explanation.

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31

5.1.2

Responsibilities of ACCs

95.
ACCs should ensure that service providers generate new animals according to CCAC

guidelines and
policies
(
see Section 4.1 and CCAC, 2003b
)
,
and
preferably

that
they
hold

a CCAC Certificate of Good
Animal Practice


GAP, or equivalent if outside of Canada.

5
.1.3

Responsibilities of Veterinarians

96. The role of the veterinarian in acquisition of animals is outlined in the
CCAC guidelines on
:

the
procurement of ani
mals

used