Iowa State University Institutional Animal Care and Use Committee (IACUC)

Effective Date: 5-17-2010 SOP ID Number: 201.03 Approved Date: 5-17-2010 Revised Date: 7-3-2019 SOP Title: Establishing Humane Endpoints Last Reviewed: 7-3-2019

Office of Research Ethics Page 1 of 14 Iowa State University

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 experienced by animal research models.

2. Scope

This SOP is applicable to all ISU staff, research investigators, and technicians who perform procedures on research, teaching and testing animals. ISU staff, research investigators, and technicians are expected to understand the content of SOPs related to their work responsibilities.

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 intervention criteria 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 for prompt identification of pain • 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.

Office of Research Ethics Page 2 of 14 Iowa State University

• 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.

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–V 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 2 cm3 for a

mouse or 5 cm3 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 “Indicators of pain” in several laboratory animal species.

Office of Research Ethics Page 3 of 14 Iowa State University

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—Rat and Mouse Appendix II. Body Condition Scores—Dogs

Appendix III. Body Condition Scores—Pigs Appendix IV. Body Condition Scores—Cows Appendix V. Body Condition Scores—Horse Appendix VI. Indicators of Pain in Several Common Laboratory Animals

9. Forms and Templates

Form I. Humane Endpoints for Laboratory Animals Template Form

Office of Research Ethics Page 4 of 14 Iowa State University

10. References

http://www.upenn.edu/regulatoryaffairs/Documents/iacuc/guidelines/iacucguideline-humaneendpointsforlaboratoryanimals.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, Iowa State University, 515-294-8507, larmedical@iastate.edu.

Office of Research Ethics Page 5 of 14 Iowa State University

Appendix I. 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 of Research Ethics Page 6 of 14 Iowa State University

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 of Research Ethics Page 7 of 14 Iowa State University

Appendix II. Body Condition Scores—Dogs

Office of Research Ethics Page 8 of 14 Iowa State University

Appendix III. Body Condition Scores—Pigs

Office of Research Ethics Page 9 of 14 Iowa State University

Appendix IV. Body Condition Scores—Cows

Office of Research Ethics Page 10 of 14 Iowa State University

Appendix IV. Body Condition Scores—Horse

From Purinamills.com website: https://www.purinamills.com/horse-feed/education/detail/body-condition-scoring-your-horse

Office of Research Ethics Page 11 of 14 Iowa State University

From Purinamills.com website: https://www.purinamills.com/horse-feed/education/detail/body-condition-scoring-your-horse

Office of Research Ethics Page 12 of 14 Iowa State University

Appendix VI. Indicators of Pain in Several Common Laboratory Animals

Species General Behavior Appearance Other

Rodents Decreased activity; excessive licking and scratching; self-mutilation; may be unusually aggressive; abnormal locomotion (stumbling, falling); writhing; does not make nest; hiding

Piloerection; rough/stained hair coat; abnormal stance or arched back; porphyrin staining (rats)

Rapid, shallow respiration; decreased food/water consumption; tremors

Rabbits Head pressing; teeth grinding; may become more aggressive; increased vocalizations; excessive licking and scratching; reluctant to locomote

Excessive salivation; hunched posture

Rapid, shallow respiration; decreased food/water consumption

Dogs Excessive licking; increased aggression; increased vocalizations, inclusive of whimpering, howling, and growling; excessive licking and scratching; self-mutilation

Stiff body movements; reluctant to move; trembling; guarding

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/iacucguideline-humaneendpointsforlaboratoryanimals.pdf

Office of Research Ethics Page 13 of 14 Iowa State University

Form I. Humane Endpoints for Laboratory Animals Template Form

Variable Score 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 1-2oC, cardiac and respiratory rates ↑up to 30% 3 Body temp change of > 2oC, 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:

Adapted from University of Pennsylvania IACUC Guidelines for Humane Endpoints Appendix C. http://www.upenn.edu/regulatoryaffairs/Documents/iacuc/guidelines/iacucguideline-humaneendpointsforlaboratoryanimals.pdf

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

Office of Research Ethics Page 14 of 14 Iowa State University

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.