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E
xperiments using laboratory animals should be well suggesting that authors publish their raw data electronically
designed, efficiently executed, correctly analyzed, (Altman 2002).
clearly presented, and correctly interpreted if they are
to be ethically acceptable. Unfortunately, surveys of pub-
lished papers reveal that many fall short of this ideal, and in
some cases, the conclusions are not even supported by the Ethical Considerations
data (Festing 1994; Festing and Lovell 1995, 1996; Mc-
The use of animals in scientific experiments likely to cause
pain, distress, or lasting harm generates important ethical
issues. Animals should be used only if the scientific objec-
Michael F. W. Festing, M.Sc., Ph.D., D.Sc., CStat., BIBiol., is a Senior
tives are valid, there is no other alternative, and the cost to
Research Scientist at the MRC Toxicology Unit, University of Leicester,
UK. Douglas G. Altman, Ph.D., D.Sc., is Director of the Cancer Research the animals is not excessive. “Validity” in this case implies
UK/NHS Centre for Statistics in Medicine, Institute of Health Sciences, that the experiment has a high probability of meeting the
Headington, Oxford, UK. stated objectives, and these objectives have a reasonable
• The objectives of the research and/or the hypotheses to Investigations Involving Several Experiments
be tested;
• The reason for choosing their particular animal model; Scientific articles often report the results of several inde-
• The species, strain, source, and type of animal used; pendent experiments. When two or more experiments are
• The details of each separate experiment being reported, presented, they should be clearly distinguished and each
including the study design and the number of animals should be described fully. It is helpful to readers to number
used; and the experiments.
• The statistical methods used for analysis.
Treatments should be assigned so that each experimental Several formal experimental designs are described in the
unit has a known, often equal, probability of receiving a literature, and most experiments should use one of these
given treatment. This process, termed randomization, is es- designs. The most common are completely randomized,
sential because there are often sources of variation, known randomized block (see below), and factorial designs; how-
or unknown, which could bias the results. Most statistical ever, Latin square, crossover, repeated measures, split-plot,
packages for computers will produce random numbers incomplete block, and sequential designs are also used.
within a specified range, which can be used in assigning These formal designs have been developed to take account
experimental units to treatments. Some textbooks have of special features and constraints of the experimental ma-
tables of random numbers designed for this purpose. Alter- terial and the nature of the investigation. It is not possible to
natively, treatment assignments can be written on pieces of describe all of the available experimental designs here. They
paper and drawn out of a bag or bowl for each experimental are described in many statistical textbooks.
unit (e.g., animal or cage). If possible, the randomization Investigators are encouraged to name and describe fully
method should ensure that there are predefined numbers in the design they used to enable readers to understand exactly
each treatment group. what was done. We also recommend including an explana-
Note that the different treatment groups should be pro- tion of a nonstandard design, if used.
cessed identically throughout the whole experiment. For Within each type of design there is considerable flex-
example, measurements should be made at the same times. ibility in terms of choice of treatments and experimental
Furthermore, animals of different treatment groups should conditions; however, standardized methods of statistical
not be housed on different shelves or in different rooms analysis are usually available. In particular, when experi-
because the environments may be different (see Blinding ments produce numerical data, they can often be analyzed
and Block Designs below). using some form of the analysis of variance (ANOVA1).
Completely randomized designs, in which animals (or
other experimental units) are assigned to treatments at ran-
Blinding dom, are widely used for animal experiments. The main
advantages are simplicity and tolerance of unequal numbers
To avoid bias, experiments should be performed “blind” in each group, although balanced numbers are less impor-
with respect to the treatments when possible and particu- tant now that good statistical software is available for ana-
larly when there is any subjective element in assessing the lyzing more complex designs with unequal numbers in each
results. After the randomized allocation of animals (or other group. However, simple randomization cannot take account
experimental unit) to the treatments, animals, samples, and of heterogeneity of experimental material or variation (e.g.,
treatments should be coded until the data are analyzed. For due to biological rhythms or environment), which cannot be
example, when an ingredient is administered in the diet, the controlled over a period of time.
different diets can be coded with numbers and/or colors and Randomized complete block designs are used to split an
the cages can be similarly coded to ensure that the correct experiment into a number of “mini-experiments” to increase
diet is given to each cage. Animals can be numbered in
random order so that at the postmortem examination there
will be no indication of the treatment group. Pathologists 1
Abbreviations used in this article: ANOVA, analysis of variance; DF,
who read slides from toxicity experiments are often not degrees of freedom.
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