Step-by-Step Three Rs Search Strategy

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A Three Rs information search typically requires a multi-database literature search, a review of CCAC guidelines & polices on animal use protocols, a search of Three Rs internet resources, and often, consultations with appropriate experts (such as laboratory animal veterinarians, animal welfare specialists and statisticians). The step-by-step strategy begins with collecting pre-search information and keyword selection, followed by two phases of information searching and review. It may be helpful to fill-out the Animal use protocol worksheet as you work through your Three Rs search.

For basic information on conducting information searches (i.e. use of operators etc.), refer to Search Basics

Step1: Collect pre-search information

Before beginning a Three Rs search, investigators should gather as much information as possible about the proposed animal use (i.e. experiment, test, training procedure). This includes:

  1. the working title of the project and the scientific objective(s);
  2. a description of the proposed animal model;
  3. descriptions of the proposed procedures on animals;
  4. descriptions of what could potentially cause pain and distress in the animals;
  5. descriptions of any known species-specific considerations;
  6. the expected Category of Invasiveness (refer to CCAC policy statement: categories of invasiveness in animal experiments);
  7. a list of any known potential Replacement alternatives (whether or not it is believed they could be useful); and
  8. a list of any known potential Reduction and/or Refinement alternatives (whether or not it is believed they could be useful).

Step 2: Select keywords and concepts

Keywords are typically found in the title, abstract and descriptor fields of a citation. In a Three Rs literature search they will relate to both the scientific topic under study (e.g. cardiology, rats) and possible alternative methods (e.g. analgesia, cellular).

Search concepts are short combinations of keywords (for example relating to the animal model or expected experimental outcome). Non-specific concepts like “animal testing alternative” and “animal use alternative” may not be useful search terms in many databases.

To generate keywords and concepts:

  1. review gathered pre-search information;
  2. consider all synonyms and different spellings of keywords(e.g., "liver cells" and "hepatocytes", “anaesthesia” and “anesthesia”, “haemoglobin” and “hemoglobin”); and
  3. review one of the following online lists of Three Rs-related search terms:

Step 3: Conduct first phase of search

The first phase is a broad search of both subject-specific information and Three Rs information, with more emphasis on Replacement alternatives. Objectives of the first phase are to:

  1. determine if the proposed experiment or test is duplicative;
  2. search specifically for Replacement alternatives (such as less sentient or non-animal methods);
  3. find information about similar scientific uses of animals (which may provide information about possible alternatives, appropriate animal numbers, and what variability to expect); and
  4. search for information that substantiates the choice of animal model.

Three Rs information may be found through the following resources:

Database resources
“Embedded” alternatives search resources

By clicking on an embedded search option, a stored Three Rs-related search will be initiated in the associated database in real time.

General resources

Step 4: Review information gathered in first phase

To evaluate the usefulness of the first phase of the search, review the relevant literature in full rather than just abstracts. This is recommended because often the information relating to Three Rs alternatives is found in the methods section of articles, not abstracts. To identify knowledge gaps that still need to be filled, it may be useful consider some or all of the Questions to assess whether your Three Rs search is complete.

Step 5: Conduct second phase of search

The second phase is a more focused search for Three Rs information, with greater emphasis on Reduction and Refinement alternatives. It also aims to fill the knowledge gaps identified in step 4. To conduct this phase:

  1. review suggestions provided for first phase of search (step 3);
  2. visit Reduction alternatives;
  3. consult a statistician as necessary;
  4. visit Refinement alternatives;
  5. review relevant CCAC guidelines; and
  6. consult a laboratory animal veterinarian or animal welfare specialist as necessary.

Step 6: Review information gathered in second phase

To evaluate the usefulness of the second phase of the search, review the relevant literature in full rather than just abstracts. This is recommended because often the information relating to Three Rs alternatives is found in the methods section of articles, not abstracts. To assist in deciding if your search is finished, it may be useful consider some or all of the Questions to assess whether your Three Rs search is complete.

Step 7: Finalize experimental plans and animal use protocol

Incorporate Three Rs alternatives into your animal-based procedures and document the Three Rs search in your animal use protocol. Double-check that your animal use protocol follows relevant CCAC guidelines & policies on animal use protocols and any policies or standard operating procedures (SOPs) at your home institution. Consider how a description of the Three Rs alternatives used could be included in any future publications about your work.

    References used in the preparation of the Three Rs Search Guide:
  • Allen T. & Jensen D. (2006) Searching bibliographic databases for alternatives. Animal Welfare Information Centre Bulletin 12(3-4): 1-2, 9-16.
  • Bottrill K. (2004) Search strategies on the internet: general and specific. ATLA 22: S585-589.
  • Focus on Alternatives (FoA) (2002) Early planning poster for a project that might involve animals. 
  • Focus on Alternatives (FoA) (2002) A general strategy for a project that might involve animals.
  • Fund for the Replacement of Animals in Medical Experiments (FRAME) (2007) How to search for information: A guide to searching for alternatives to the use of laboratory animals. 
  • Grune B., Fallon M., Howard C., Hudson V., Kulpa-Eddy J., Larson J., Leary S., Roi A., van der Valk J., Wood M., Dorendahl A., Kohler-Hahn D., Box R. & Spielman H. (2004) Report and recommendations of the international workshop "Retrieval approaches for information on alternative methods to animal experiments". ALTEX 21: 115-127.
  • Hakkinen P. & Green D. (2002) Alternatives to animal testing: information resources via the internet and world wide web. Toxicology 173: 3-11.
  • Hart L., Wood M. & Weng H.-Y. (2005) Effective searching of the scientific literature for alternatives: search grids for appropriate databases. Animal Welfare 14: 287-289.
  • Jensen D. (2005) Worksheet and instructions for alternatives literature searching. Animal Welfare Information Center. Beltsville MD: USDA.
  • Jensen D. (2006) An alternatives search example for advanced trauma life support training. Animal Welfare Information Centre Bulletin 12(3-4): 3-5.
  • Kilkenny C., Browne W.J., Cuthill I.C., Emerson M. and Altman D.G. (2010) Improving Bioscience Research Reporting: The ARRIVE Guidelines for Reporting Animal Research. PLoS Biology 8(6), (accessed on 2010-09-14).
  • Langley G., Broadhead C., Bottrill K., Combes R., Ewbank R., Hawkins P., Hubrecht R., Jennings M., Newman C., Rowe S., Southee J., Todd M. & Ward L. (1999) Accessing information on the reduction, refinement and replacement of animal experiments: Report and recommendations of a Focus on Alternatives workshop. ATLA 27: 239-245.
  • Smith A. & Allen T. (2005) The use of databases, information centres and guidelines when planning research that may involve animals. Animal Welfare 14: 347-359.
  • Wood M. (2005) Search for alternatives: A step-by-step approach to alternatives searches. University of California Centre for Animal Alternatives. 
Last modified: 2010-11-17