The principles of the 3 Rs are also reflected in the Guide, which says that the following topics should be considered in the development and review of animal protocols (p. 10):

  • Justification of the species and number of animals requested.

  • Availability or appropriateness of the use of less-invasive procedures, other species, isolated organ preparation, cell or tissue culture, or computer simulation.

  • Appropriate sedation, analgesia, and anesthesia.

  • Unnecessary duplication of experiments.

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