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Appendix
Animal Research: Reporting In Vivo Experiments:
The ARRIVE Guidelines7
ITEM RECOMMENDATION
TITLE 1 Provide as accurate and concise a description of the content
of the article as possible.
ABSTRACT 2 Provide an accurate summary of the background, research
objectives, including details of the species or strain of animal
used, key methods, principal findings and conclusions of the
study.
INTRODUCTION
Background 3 a. Include sufficient scientific background (including relevant
references to previous work) to understand the motivation
and context for the study, and explain the experimental
approach and rationale.
b. Explain how and why the animal species and model being
used can address the scientific objectives and, where
appropriate, the study’s relevance to human biology.
Objectives 4 Clearly describe the primary and any secondary objectives of
the study, or specific hypotheses being tested.
METHODS
Ethical statement 5 Indicate the nature of the ethical review permissions,
relevant licences (e.g. Animal [Scientific Procedures] Act
1986), and national or institutional guidelines for the care
and use of animals, that cover the research.
Study design 6 For each experiment, give brief details of the study design
including:
a. The number of experimental and control groups.
b. Any steps taken to minimise the effects of subjective bias
when allocating animals to treatment (e.g. randomisation
procedure) and when assessing results (e.g. if done, describe
who was blinded and when).
c. The experimental unit (e.g. a single animal, group or cage
of animals).
A time-line diagram or flow chart can be useful to illustrate
how complex study designs were carried out.
Experimental procedures 7 For each experiment and each experimental group, including
controls, provide precise details of all procedures carried out.
For example:
a. How (e.g. drug formulation and dose, site and route of
administration, anaesthesia and analgesia used [including
monitoring], surgical procedure, method of euthanasia).
Provide details of any specialist equipment used, including
7
Reprinted with permission from the UK National Centre for the Replacement, Refinement and Reduction of
Animals in Research (www.nc3rs.org.uk). Originally published in PLoS Biology, June 2010 (volume 8, issue 6).
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supplier(s).
b. When (e.g. time of day).
c. Where (e.g. home cage, laboratory, water maze).
d. Why (e.g. rationale for choice of specific anaesthetic, route
of administration, drug dose used).
Experimental animals 8 a. Provide details of the animals used, including species,
strain, sex, developmental stage (e.g. mean or median age
plus age range) and weight (e.g. mean or median weight plus
weight range).
b. Provide further relevant information such as the source of
animals, international strain nomenclature, genetic
modification status (e.g. knock-out or transgenic), genotype,
health/immune status, drug or test naïve, previous
procedures, etc.
Housing and husbandry 9 Provide details of:
a. Housing (type of facility e.g. specific pathogen free [SPF];
type of cage or housing; bedding material; number of cage
companions; tank shape and material etc. for fish).
b. Husbandry conditions (e.g. breeding programme,
light/dark cycle, temperature, quality of water etc for fish,
type of food, access to food and water, environmental
enrichment).
c. Welfare-related assessments and interventions that were
carried out prior to, during, or after the experiment.
Sample size 10 a. Specify the total number of animals used in each
experiment, and the number of animals in each experimental
group.
b. Explain how the number of animals was arrived at. Provide
details of any sample size calculation used.
c. Indicate the number of independent replications of each
experiment, if relevant.
Allocating animals to experimental 11 a. Give full details of how animals were allocated to
groups experimental groups, including randomisation or matching if
done.
b. Describe the order in which the animals in the different
experimental groups were treated and assessed.
Experimental outcomes 12 Clearly define the primary and secondary experimental
outcomes assessed (e.g. cell death, molecular markers,
behavioural changes).
Statistical methods 13 a. Provide details of the statistical methods used for each
analysis.
b. Specify the unit of analysis for each dataset (e.g. single
animal, group of animals, single neuron).
c. Describe any methods used to assess whether the data
met the assumptions of the statistical approach.
RESULTS
Baseline data 14 For each experimental group, report relevant characteristics
and health status of animals (e.g. weight, microbiological
status, and drug or test naïve) prior to treatment or testing.
(This information can often be tabulated).
Numbers analysed 15 a. Report the number of animals in each group included in
each analysis. Report absolute numbers (e.g. 10/20, not
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50%2).
b. If any animals or data were not included in the analysis,
explain why.
Outcomes and estimation 16 Report the results for each analysis carried out, with a
measure of precision (e.g. standard error or confidence
interval).
Adverse events 17 a. Give details of all important adverse events in each
experimental group.
b. Describe any modifications to the experimental protocols
made to reduce adverse events.
DISCUSSION
Interpretation/scientific implications 18 a. Interpret the results, taking into account the study
objectives and hypotheses, current theory and other relevant
studies in the literature.
b. Comment on the study limitations including any potential
sources of bias, any limitations of the animal model, and the
imprecision associated with the results†.
c. Describe any implications of your experimental methods or
findings for the replacement, refinement or reduction (the
3Rs) of the use of animals in research.
Generalisability/translation 19 Comment on whether, and how, the findings of this study are
likely to translate to other species or systems, including any
relevance to human biology.
Funding 20 List all funding sources (including grant number) and the role
of the funder(s) in the study.
The guidelines are intended to:
— Improve reporting of research using animals.
— Guide authors as to the essential information to include in a manuscript, and not be absolutely
prescriptive.
— Be flexible to accommodate reporting a wide range of research areas and experimental
protocols.
— Promote reproducible, transparent, accurate, comprehensive, concise, logically ordered, well
written manuscripts.
— Improve the communication of the research findings to the broader scientific community.
The guidelines are NOT intended to:
— Promote uniformity, stifle creativity, or encourage authors to adhere rigidly to all items in the
checklist. Some of the items may not apply to all studies, and some items can be presented as
tables/figure legends or flow diagrams (e.g. the numbers of animals treated, assessed and
analysed).
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— Be a guide for study design and conduct. However, some items on the checklist, such as
randomisation, blinding and using comparator groups, may be useful when planning
experiments as their use will reduce the risk of bias and increase the robustness of the research.
What kind of research areas do the guidelines apply to?
— The guidelines will be most appropriate for comparative studies, where two or more groups of
experimental animals are being compared; often one or more of the groups may be considered
as a control. They apply also to studies comparing different drug doses, or, for example, where a
single animal is used as its own control (within–subject experiment).
— Most of the recommendations also apply to studies that do not have a control group.
— The guidelines are suitable for any area of bioscience research where laboratory animals are
used.
Who are the guidelines aimed at?
— Novice and experienced authors
— Journal editors
— Peer reviewers
— Funding bodies
How might these guidelines be used?
The guidelines provide a checklist for those preparing or reviewing a manuscript intended for
publication.
References
1. Kilkenny C, Browne WJ, Cuthill IC, Emerson M, Altman DG (2010) Improving Bioscience Research
Reporting: The ARRIVE Guidelines for Reporting Animal Research. PLoS Biol 8(6): e1000412.
doi:10.1371/journal.pbio.1000412
2. Schulz KF, Altman DG, Moher D, the CONSORT Group (2010) CONSORT 2010 Statement: updated
guidelines for reporting parallel group randomised trials. BMJ 340:c332.
Acknowledgements
The NC3Rs gratefully acknowledges the expertise and advice that all the contributors have given to
developing the guidelines. We would particularly like to acknowledge the contribution of the NC3Rs
Reporting Guidelines Working Group† – Professor Doug Altman, Centre for Statistics in Medicine,
University of Oxford UK, Professor David Balding, Department of Epidemiology & Public Health, Imperial
College, London UK, Professor William Browne, Department of Clinical Veterinary Science, University of
Bristol UK, Professor Innes Cuthill, School of Biological Sciences, University of Bristol UK, Dr Colin Dunn,
Editor Laboratory Animals (Royal Society of Medicine press), Dr Michael Emerson, National Heart and
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Lung Institute, Imperial College, London UK, Dr Stella Hurtley, Senior Editor Science, Professor Ian
McGrath, Editor-in-Chief British Journal of Pharmacology (Wiley Blackwell Publishers) and Dr Clare
Stanford, Department of Psychopharmacology, University College, London UK. We would also like to
thank NC3Rs grant holders, the Medical Research Council, Biotechnology and Biological Sciences
Research Council (BBSRC), Wellcome Trust, Parkinson’s Disease Society, British Heart Foundation and
their grant holders and funding committee members who provided feedback on the guidelines; and
Kathryn Chapman and Vicky Robinson (both NC3Rs) for their help with the manuscript.
†Please note that the working group members who contributed to these guidelines were advising in
their personal capacity and their input does not necessarily represent the policy of the organisations
with which they are associated.
Funding
The reporting guidelines project was funded by the National Centre for the Replacement, Refinement
and Reduction of Animals in Research (NC3Rs).
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