Effective Date: 5-17-2010 Approved Date: 5-17-2010 Revised Date: 5-11-2016 Last Reviewed: 5-11-2016 Institutional Animal Care and Use Committee (IACUC) SOP ID Number: 201.02 SOP Title: Establishing Humane Endpoints Institutional Animal Care and Use Committee (IACUC) Standard Operating Procedure ESTABLISHING HUMANE ENDPOINTS 1. Purpose The purpose of this SOP is to provide instruction on how to set objective, humane interventions for pain for animal research models. 2. Scope PIs and their research staff, LAR staff, and ISU IACUC. 3. General Requirements In using animals for research or teaching, one must consider the possibility that pain or distress may exceed what is anticipated. Humane interventions are clear criteria to prevent or alleviate unnecessary pain or distress in a research or teaching animal. Humane interventions may include but are not limited to the following: Adequate veterinary care, analgesia, and/or supportive care to the animal Modification to housing or husbandry practices to improve the comfort of the animal Increasing the frequency of animal observations Modification of the experimental procedures to decrease the discomfort to the animals Termination of painful procedures Removal of animals from the study Humane euthanasia of the animal Pilot studies should be used to determine intervention criteria where the effects of the disease or study is not known. Ensuring humane intervention criteria is a responsibility of the PI, the veterinary care staff, and the IACUC: Determine intervention criteria that are appropriate for the study. Ensure that humane intervention criteria are clearly defined in the IACUC protocol. Ensure that all personnel responsible for making animal observations have been trained to observe, recognize, and document clinical criteria and then respond appropriately to the intervention criteria established in the approved IACUC protocol. Office for Responsible Research Page 1 of 12
4. Procedure Establishing Intervention Criteria: Review literature and perform internet search of established models and alternatives. Consult with veterinary care staff on study refinements to minimize pain and distress. Schedule regular animal observations performed by the research staff and include this schedule in the IACUC protocol. Keep records of all observations including specific measurements or data. General Intervention Criteria: Weight loss exceeding 20% of baseline bodyweight. For growing animals, failure to maintain normal weight gain within 15% of age-matched control animals. Body Condition Score (BCS) of less than 2 (See Appendices I IV for BCS charts of common laboratory animals.) Uncontrollable seizures, incoordination, or paralysis Hunched posture, lethargy, and lack of grooming Decreased mobility which interferes with normal eating, drinking, ambulating, or grooming Respiratory distress, labored breathing, increased or decreased respiratory rate, cyanosis Uncontrolled bleeding Self-mutilation Specific organ failure Mass that is ulcerated, necrotic, impairing function, or exceeding 2cm 3 for a mouse or 5cm 3 for a rat Use of objective, measurable parameters whenever possible Creation of score sheets or assessment checklists Refer to the appendices in this document for body condition scores (BCS), a sample scoring form, and a table of indicators of pain in several laboratory animal species. Office for Responsible Research Page 2 of 12
5. Roles and Responsibilities Roles Responsibilities PIs Review the literature and determine possible adverse effects and criteria for assessment of pain and distress. Consult with LAR veterinarians for possible study refinements to minimize pain and distress. Schedule animal observations at an appropriate frequency. Ensure individuals assessing animals have prior appropriate training. LAR Veterinary Staff Provide clinical evaluation of animals on study. Provide guidance and consultation to researchers in choosing humane intervention criteria and endpoints. Document relevant clinical details in medical records. Apply IACUC approved interventions in consultation with the relevant research personnel. IACUC Members Review humane intervention criteria in protocols. Approve appropriate intervention criteria in protocols. Refer the Principal Investigator to LAR veterinary staff if humane intervention criteria in an IACUC protocol needs revision prior to approval. 6. Definitions Body Condition Scores (BCS): A management tool for distinguishing differences in body condition of animals which uses a numeric score to estimate body condition. 7. Health and Safety Information n/a 8. Appendices Appendix I. Body Condition Scores Dogs Appendix II. Body Condition Scores Pigs Appendix III. Body Condition Scores Cows Appendix IV. Body Condition Scores Rat and Mouse Appendix V. Indicators of Pain in Several Common Laboratory Animals 9. Forms and Templates Humane Endpoints for Laboratory Animals Template Form Office for Responsible Research Page 3 of 12
10. References http://www.upenn.edu/regulatoryaffairs/documents/iacuc/guidelines/iacucguidelinehumaneendpointsforlaboratoryanimals.pdf https://awic.nal.usda.gov/humane-endpoints-and-euthanasia http://ilarjournal.oxfordjournals.org/content/41/2.toc https://www.humane-endpoints.info/en http://altweb.jhsph.edu/pubs/proceedings/pain/proceedings.html http://www.ccac.ca/documents/standards/guidelines/appropriate_endpoint.pdf http://oacu.od.nih.gov/arac/documents/asp_endpoints.pdf Why Do a Pilot Study? National Center for the Replacement, Refinement and Reduction of Animals in Research. http://nc3rs.org.uk/page.asp?id=275 Sneddon, Lynne U. 2009. Pain perception in fish: indicators and endpoints. ILAR Journal 50, no. 4: 338 342. http://journals.plos.org/plospathogens/article?id=10.1371/journal.ppat.1002399 Dunlap, Jeremiah. 2015. Humane endpoints for animals used in training. Lab Animal 44.2: 71 71. Miller, S., et al. 2012. Mouse traumatic brain injury model: Clinical care, humane endpoints, and neurologic assessment. Journal of the American Association for Laboratory Animal Science Vol. 51, no. 5. 11. Contact Information Contact the Attending Veterinarian with questions: Dr. Mary Sauer, VMD, 515-294-0266, msauer@iastate.edu or LAR Medical, Laboratory Animal Resources,, 515-294-8507, larmedical@iastate.edu. Office for Responsible Research Page 4 of 12
Appendix I. Body Condition Scores Dogs Office for Responsible Research Page 5 of 12
Appendix II. Body Condition Scores Pigs Office for Responsible Research Page 6 of 12
Appendix III. Body Condition Scores Cows Office for Responsible Research Page 7 of 12
Appendix IV. Body Condition Scores Rat and Mouse Charmain J Foltz. 1999. Body Condition Scoring: A Rapid and Accurate Method for Assessing Health Status in Mice. Lab. Animal Science Vol 49 (3) 319 323. Office for Responsible Research Page 8 of 12
Hickman, D. L., M. Swan. 2010. Use of a body condition score technique to assess health status in a rat model of polycystic kidney disease. J Am Assoc Lab Anim Sci 49(2):155 9. Office for Responsible Research Page 9 of 12
Appendix V. Indicators of Pain in Several Common Laboratory Animals Species General Behavior Appearance Other Rodents Rabbits Dogs Decreased activity; excessive licking and scratching; selfmutilation; may be unusually aggressive; abnormal locomotion (stumbling, falling); writhing; does not make nest; hiding Head pressing; teeth grinding; may become more aggressive; increased vocalizations; excessive licking and scratching; reluctant to locomote Excessive licking; increased aggression; increased vocalizations, inclusive of whimpering, howling, and growling; excessive licking and scratching; selfmutilation Piloerection; rough/stained hair coat; abnormal stance or arched back; porphyrin staining (rats) Excessive salivation; hunched posture Stiff body movements; reluctant to move; trembling; guarding Rapid, shallow respiration; decreased food/water consumption; tremors Rapid, shallow respiration; decreased food/water consumption Decreased food/water consumption; increased respiration rate/panting Adapted from University of Pennsylvania IACUC Guidelines for Humane Endpoints Appendix A. http://www.upenn.edu/regulatoryaffairs/documents/iacuc/guidelines/iacucguidelinehumaneendpointsforlaboratoryanimals.pdf Office for Responsible Research Page 10 of 12
Humane Endpoints for Laboratory Animals Template Form Variable Value Body Weight Changes 0 Normal 1 < 10 percent weight loss 2 10-15 percent weight loss 3 > 20 percent weight loss Value Body Condition Score (BCS, see diagram examples) 0 BCS >3 1 BCS >2 and < 3 2 BCS >1 and 3 BCS of 1 or less Value Physical Appearance 0 Normal 1 Lack of grooming 2 Rough coat, nasal/ocular discharge 3 Very rough coat, abnormal posture, enlarged pupils Value Measurable Clinical Signs 0 Normal 1 Small changes of potential significance 2 Body temp change of 1-2 o C, cardiac and respiratory rates up to 30% 3 Body temp change of > 2 o C, cardiac and respiratory rates up to 50%, or markedly reduced Value Unprovoked Behavior 0 Normal 1 Minor changes 2 Abnormal, reduced mobility, decreased alertness, inactive 3 Unsolicited vocalizations, self-mutilation, either very restless or Value Behavioral Responses to External Stimuli 0 Normal 1 Minor depression/exaggeration of response 2 Moderately abnormal responses 3 Violent reactions, or comatose TOTAL: Score Adapted from University of Pennsylvania IACUC Guidelines for Humane Endpoints Appendix C. http://www.upenn.edu/regulatoryaffairs/documents/iacuc/guidelines/iacucguidelinehumaneendpointsforlaboratoryanimals.pdf Office for Responsible Research Page 11 of 12
Note: This scoring template should be modified for specific species and designed to fit each protocol and animal model. In this example, a score is assigned to each variable, 0 (normal or mild) to 3 (severe). The cumulative score gives an indication of the likelihood that the animal is experiencing pain or distress. Humane endpoints can be established based on these criteria. A total score of >5 or a score of 3 in any one variable, regardless of the total score, should warrant mandatory evaluation/decision by a veterinarian or humane euthanasia. Office for Responsible Research Page 12 of 12