Table Of ContentThe COST Manual of
Laboratory Animal Care and Use
Refinement, Reduction, and Research
The COST Manual of
Laboratory Animal Care and Use
Refinement, Reduction, and Research
Edited by
Bryan Howard
Timo Nevalainen
Gemma Perretta
Boca Raton London New York
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Contents
COST and COST Action B-24 .........................................................................................................vii
Preface...............................................................................................................................................ix
Editors ...............................................................................................................................................xi
Contributors ...................................................................................................................................xiii
Editorial Board of the Manual .......................................................................................................xvii
Chapter 1 Introduction ..................................................................................................................1
Bryan Howard, United Kingdom
Chapter 2 Design and Oversight of Laboratory Animal Facilities ...............................................7
Dag Sørensen, Norway; Heinz Brandstetter, Germany;
Nikolaos Kostomitsopoulos, Greece; and Richard Fosse, France
Chapter 3 Housing and Care of Laboratory Animals .................................................................29
Hanna-Marja Voipio, Finland; Ping-Ping Tsai, Germany; Heinz Brandstetter,
Germany; Marcel Gyger, Switzerland; Hansjoachim Hackbarth, Germany;
Axel Kornerup Hansen, Denmark; and Thomas Krohn, Denmark
Chapter 4 Animal Needs and Environmental Refinement .........................................................75
Vera Baumans, The Netherlands; Hanna Augustsson, Sweden;
and Gemma Perretta, Italy
Chapter 5 Ethical Evaluation of Scientific Procedures: Recommendations for Ethics
Committees ..............................................................................................................101
Rony Kalman, Israel; I. Anna S. Olsson, Portugal; Claudio Bernardi,
Italy; Frank van den Broek, The Netherlands; Aurora Brønstad, Norway;
Istvan Gyertyan, Hungary; Aavo Lang, Estonia; Katerina Marinou,
Greece; and Walter Zeller, Switzerland
Chapter 6 Reduction by Careful Design and Statistical Analysis ............................................131
Michael Festing, United Kingdom
Chapter 7 Animal Models: Selecting and Preparing Animals for a Study...............................151
Patrick Hardy, France and Sarah Wolfensohn, United Kingdom
Chapter 8 Creation of Genetically Modified Animals ..............................................................179
Belen Pintado, Spain and Marian van Roon, The Netherlands
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Chapter 9 Management of Genetically Modified Rodents .......................................................205
Jan-Bas Prins, The Netherlands
Chapter 10 Impact of Handling, Radiotelemetry, and Food Restriction ....................................227
Timo Nevalainen, Finland: Marlies Leenaars, The Netherlands;
Vladiana Crljen, Croatia; Lars Friis Mikkelsen, Denmark; Ismene Dontas,
Greece; Bart Savenije, The Netherlands; Carlijn Hooijmans, The Netherlands;
and Merel Ritskes-Hoitinga, The Netherlands
Chapter 11 Basic Procedures: Dosing, Sampling and Immunisation .........................................257
Ismene Dontas, Greece; Jann Hau, Denmark; Katerina Marinou, Greece;
and Timo Nevalainen, Finland
Chapter 12 Imaging Techniques .................................................................................................287
Aurora Brønstad, Norway and Ismene Dontas, Greece
Chapter 13 Anaesthesia and Analgesia .......................................................................................313
Patricia Hedenqvist, Sweden and Paul Flecknell, United Kingdom
Chapter 14 Use of Humane Endpoints to Minimise Suffering ...................................................333
Coenraad Hendriksen, The Netherlands; David Morton, United Kingdom;
and Klaus Cussler, Germany
Chapter 15 Euthanasia ................................................................................................................355
Luis Antunes, Portugal
Chapter 16 Education, Training, and Competence .....................................................................369
Bryan Howard, United Kingdom; Katey Howard, Spain; and Peter Sandøe,
Denmark
Chapter 17 Animal Experimentation and Open Communication ..............................................391
Ann-Christine Eklöf, Sweden; Anne-Grethe Berg, Norway;
and Jon Richmond, United Kingdom
Index ..............................................................................................................................................405
COST and COST Action B-24
COST is an intergovernmental framework for European Cooperation in Science and Technology,
which promotes and coordinates nationally funded research in Europe. COST provides funds for
research networks (Actions) and in this way helps consolidate European research investment and
opens the European research area to cooperation worldwide.
COST is solely funded from a specific part of the EU RTD Framework Programmes. The finan-
cial support that COST provides to Actions is used for coordination and networking activities, while
the research itself is funded at the national/EU level. In this way, COST reaches out to over 30,000
researchers across Europe and levers approximately EUR 2 billion of research funding, although its
direct support is less than 1% of this sum.
One of the key scientific domains that COST covers (out of a total of nine) is the Domain of
Biomedicine and Molecular Biosciences (BMBS), which at the time of writing (June 2010) is
funding a total of 29 Actions.
This manual summarises the output, over a period of four years, of BMBS COST Action
B-24 on “Laboratory Animal Science and Welfare”. The remit of Action B-24 was to increase
knowledge and awareness of the scientific uses of laboratory animals within the context of the
Three Rs, and to promote the conduct of high-quality research. The Action began its activi-
ties in March 2004 and came to an end in April 2009. However, many participants have links
with national associations and with the Federation of European Laboratory Animal Science
Associations (FELASA) with which the Action has collaborated closely throughout. It is antici-
pated that FELASA will continue to serve as a coordinating body for the progress of labora-
tory animal science and the Three Rs within Europe. Twenty-four countries participated in the
Action, as shown in Table 0.1.
Participants of Action B-24 have published many papers in specialist journals and offer this man-
ual, as a collated source of up-to-date information about ethically sound approaches to the care and
scientific use of animals. It draws heavily from the widely accepted principles of Russell and Burch,
usually known as the Three Rs. We hope that this volume will be of value to those who wish to
benchmark or improve practices relating to the scientific use of animals, and will serve as a conve-
nient source of evidence-based information about good practices and laboratory animal science and
welfare to all those engaged in this field, including educators and regulatory/oversight authorities.
For more information: http://www.cost.eu
TABLE 0.1
Nations Participating in Cost Action B-24
Austria Germany The Netherlands
Belgium Greece Norway
Croatia Hungary Portugal
Czech Republic Ireland Spain
Denmark Israel Sweden
Estonia Italy Switzerland
Finland Lithuania Turkey
France Malta United Kingdom
Dr. Kalliopi Kostelidou
Cluster Leader, Cluster of Life Sciences
Science Officer, BMBS Domain
vii
Preface
A revised Appendix A to the European Convention ETS 123 came into force in 2007. This
represented a significant change to baseline standards for the care of laboratory animals within
Europe. Subsequent adoption of these guidelines by the European Commission and development
of a new Directive to replace 86/609, marks greater emphasis on the Three Rs in laboratory animal
science.
In 2004, COST (European Cooperation in Science and Technology) established Action B-24
(Laboratory Animal Science and Welfare) as an initiative to increase knowledge necessary for the
ethically and scientifically sustainable use of laboratory animals in research. The Action attracted
24 Member States, each represented by one or two members, who contributed to on-going dialogue
and research about the most humane and effective way of using laboratory animals for scientific
purposes. One result of this initiative has been the preparation of this text—The COST Manual of
Laboratory Animal Care and Use: Refinement, Reduction, and Research. The manual is a joint
effort involving many authors and it represents a truly international perspective on best practice,
as currently perceived. Although 15 different COST countries have each contributed one or more
authors in preparation of the text, the information contained includes a great deal of less formal
input by other members of the COST Action. Although it is not possible to recognise everybody
who has participated in preparation of the manual, the editors are extremely grateful for the support
received from them and also from a number of colleagues who, though not directly related to the
Action, nonetheless contributed material where specific expertise was not immediately available.
The manual can be viewed as an international venture, made possible by the funding and support
of COST.
The reason for not addressing “Replacement” in this manual is that the underlying technologies
involve different scientific disciplines and often require specialisms that are beyond the reach of
laboratory animal scientists. Omission in no way reflects a view of the editors that Replacement is
in any way less of a priority in laboratory animal science than Reduction and Refinement. Indeed,
we firmly believe that Replacement is the ultimate objective for laboratory animal science, although
we are of the opinion that science should not be sidestepped simply because appropriate replace-
ment techniques are not yet currently available. In circumstances where no replacement strategy is
capable of generating the scientific information necessary to test the hypothesis being addressed
and where the work proposed can be shown to have important applications, then every measure
needs to be taken to minimise the number of animals used and the impact of procedures on each
of those animals.
Only the more commonly used laboratory animal species are considered: rodents and rabbits.
Our intention is to offer a one-stop source of best practice, which will be of value to personnel
responsible for the care and welfare of animals and scientists conducting activities related to the
use of animals for scientific purposes. It is also addressed to those with management responsibili-
ties including facility engineers, architects and lay persons involved with ethical review, and the
interested general reader.
The manual presents perceived best practices and, as far as possible, the contents are evidence-
based. In the many cases where the evidence is poor, the authors have endeavoured to demonstrate
the rationale underlying their contentions by including relevant references. The emphasis is not on
describing current practices, but rather what is seen as good practice; moreover it is not our intention
to tell the reader what to do, but rather to offer options and provide advice on what not to do.
Each chapter follows a similar structure, comprising six sections, the first of which, “Objectives”,
outlines the breadth of what the chapter is addressing. This is followed by “Key Factors”, which
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