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Guidelines for pain and distress in laboratory animals:

Guidelines for Pain and Distress in Laboratory Animals:
Responsibilities, Recognition and Al eviation


Animals can experience pain and distress. It is the ethical and legal obligation of all personnel involved with the use of animals in research to reduce or eliminate pain and distress in research animals whenever such actions do not interfere with the research objectives. The Institutional Animal Care and Use Committee (IACUC) has the delegated responsibility and accountability for ensuring that al animals under their oversight are used humanely and in accordance with a number of Federal Regulations and policies.2,21,29,30,32 Key to fulfilling the responsibilities for both the principal investigator (PI) and the • understanding the legal requirements, • being able to distinguish pain and distress in animals from their normal state, and • to relieve or minimize the pain and distress appropriately;
Regulatory Requirements
The IACUC must assure that all aspects of the animal study proposal (ASP) that may cause more than transient pain and/or distress are addressed; alternatives6 to painful or distressful procedures are considered; and that methods, anesthetics and analgesics to minimize or eliminate pain and distress are included when these methods do not interfere with the research objectives; and that humane end points have been established for al situations where more than transient pain and distress can not be avoided or eliminated. Whenever possible, death or severe pain and distress should be avoided as end points. A written scientific justification is required to be included in the ASP for any more than transient painful or distressful procedure that cannot be relieved or minimized. The obligation to reduce pain and distress does not end with the review of the ASP. It is the
responsibility of the animal care staff, the research staff, veterinarians and the IACUC, to continue to monitor animals for pain, distress, il ness, morbidity or mortality during the course of the research study. If unexpected pain or distress occurs, and is more than an isolated incident, the PI must submit an amendment delineating the unexpected problem and stating their proposed resolution to the issue (e.g. administration of analgesics, lowering the dose of a drug that was administered, etc.). Alternatively, the PI could justify the need for unrelieved pain or distress in the amendment, or in the case of regulated If it is necessary to make changes in the ASP procedures, the PI must submit an amendment to the IACUC and receive approval prior to instituting the modification. For example, if unexpected pain or distress was noted following treatment with a specific dose of an experimental agent, the PI must submit an amendment delineating the unexpected problem and stating their proposed resolution to the issue (e.g. administration of analgesics, lowering the dose administered, etc.). Alternatively, the PI could justify the need for unrelieved pain or distress in the amendment, or in the case of regulated species as
Recognition of Pain and Distress
Animals must be monitored by trained individuals for pain and distress as appropriate for the species, condition and procedure. Critical to the assessment of the presence or absence of pain or distress is having the ability to distinguish between normal and abnormal animal behavior. This is especially true when dealing with species that often exhibit pain and distress with only subtle changes in their behavior22 (see Table 1). Therefore, it is critical that the individuals assessing an animal be trained in the species specific signs of pain and distress, as well as be knowledgeable of the potential outcomes of the procedure, surgery or treatments administered to the animal. Pain and distress scoring is a method to convert subjective animal observations in to objective scoring system which some have found to be helpful in assessing animal behavior. Whenever more than transient pain or distress is anticipated preemptive measures should be taken to minimize or prevent the development of pain and/or distress. Following the implementation of preemptive or palliative measures, animals must be monitored1,26 to ensure the efficacy of the measures taken and determine if or when additional treatment wil be necessary. The extent and frequency of monitoring will depend on the level of post-surgical/procedural pain and/or distress anticipated and the chosen intervention strategy(s). For example, animals undergoing a procedure known to produce no more than minimal/transient pain or distress may be adequately monitored by the daily observation of a trained animal caretaker. Whereas, the monitoring of an animal undergoing a procedure known to result in severe pain and/or distress may require more frequent monitoring by a team of trained individuals (e.g. trained animal care staff, technicians, veterinarians, investigators, etc.). Animals undergoing pilot studies or procedures new to the investigator or facility may also require a higher frequency of monitoring and a team approach. It is ultimately the responsibility of the PI and the personnel conducting the procedure to ensure the timely and adequate identification, monitoring and documentation of the animals undergoing potential y painful or distressful procedures. Investigators may request the assistance of institute and facility veterinary and technical personnel when monitoring their animals, but al individual(s) responsible for monitoring an animal must be identified prior to conducting the procedure and their accountability clearly delineated and accepted. Animals should be observed a minimum of once daily or more often based on professional judgment and the research being conducted. The animals should be monitored for expected and unexpected signs of more than transient pain or distress and, if observed, appropriate intervention strategies implemented (e.g. non-pharmacological approaches, analgesics, anesthetics, euthanasia, etc.), unless the withholding of treatment is scientific justified. Observations and actions taken to relieve pain or The documentation of monitoring of the animal is important and required. The nature and the frequency of the documentation are dependent on the species and the potential for pain and/or distress. For example, the identification of cages containing an animal which has under gone a potentially painful or distressful procedure with a “special observation” cage card has proven helpful in drawing special attention to the animal during the caretaker’s daily health check. Cages containing animals requiring more intensive monitoring should also be appropriately identified and their monitoring and/or treatments documented either at the room, cage or animal level (e.g. room log, cage card, medical record, etc.), in addition to the investigator’s notations in their laboratory notebook. Documentation must be available to al personnel monitoring the cage or animal (i.e. IACUC, veterinarians, animal care staff, etc.).
Intervention Strategies
Strategies for the management of pain and distress may include non pharmacological considerations (e.g. modified housing and husbandry practices, dietary modifications, surgical approaches8, desensitization and acclimation strategies, etc.), pharmacological interventions or euthanasia. The chosen strategy wil vary with the species, the procedure(s) being performed, duration of action needed, route of administration preferred, degree and type of analgesia required and research being conducted (see Table 2). It is strongly suggested that PIs consult their IC veterinarian during the development of an animal study protocol, prior to its submission to the IACUC. This approach has been demonstrated to Excellent resources3,4 and formularies7,10,12,14,15 are now available which provide extensive information on the recognition and al eviation of pain and distress in laboratory animals (See Plumb Veterinary Drug Handbook26 and other references below). These resources, coupled with trained and skilled animal care personnel and the professional judgment of your IC and animal facility veterinarian provides each investigator and their IACUC with powerful tools for the recognition and alleviation of pain and distress in laboratory animals. Preemptive measures should be taken to minimize or prevent the development of pain and/or distress. For example, a skilled surgeon can often minimize tissue trauma which in turn minimizes post- operative pain and distress. The use of ketamine or opioids preemptively, even in low doses has been demonstrated to prevent the development of some forms of pain.24 In addition, the use of a single dose of a non-steroidal anti-inflammatory agent (NSAID), sustained-release formulations,13 or other analgesic agent can have a positive effect on the speed with which animals return to normal behavior.16,17,36,37,38 It has been repeatedly demonstrated in humans that the provision of effective analgesia reduces the time There are also many pharmacological intervention strategies for the management of pain and distress. Traditional analgesics include local or regional anesthetics, opioids and NSAIDS. Using two or more of classes of these analgesics together, or combining these analgesics with nontraditional analgesics such as NMDA antagonists,20,40 alpha2-agonists,35 tramadol,5 and even the antiepileptic drug gabapentin27 has been shown in both human and veterinary patients to enhance analgesia and allow a reduction in the use of more powerful analgesics. This approach, cal ed multimodal analgesia also has the advantage of providing even analgesic dosing thus promoting the animal’s well being. For procedures in which the pain intensity is anticipated to be high, techniques such as constant rate infusions of local anesthetics and or opioids either systemically, locally at the surgical site or via an epidural catheter and transdermal preparations of drugs provide uninterrupted analgesia and are being used successfully in larger laboratory animals.9 The analgesic regimen chosen should always be made in The relief of pain and distress in research animals is ethical y sound, humane, and promotes good science. The establishment of clear lines of responsibility coupled with appropriate endpoints, monitoring and intervention strategies are key to the prevention, minimization and/or al eviation of pain and distress in laboratory animals. Several excellent references and formularies are available to the researcher, veterinarian and husbandry personnel to facilitate their ability to recognize and modulate pain and distress in laboratory animals. Experience has demonstrated that a dynamic, interactive team approach to the recognition and alleviation of pain and distress in laboratory animals yields results that protect animal-welfare while promoting good science. Revised - 07/14/04; 05/16/07; 07/14/10; 11/14/12 References
1. Adamson, TW, Kendall, LV, Goss, S, et al. 2010 Assessment of Carprofen and Buprenorphine on Recovery of Mice after Surgical Removal of the Mammary Fat Pad. JAALAS 49:(5) 610-616. 2. Animal Welfare Act: Public Law 89-544, 1966, as amended, (P.L. 91-579, P.L. 94 -279 and P.L. 99-198) 7 U.S.C. 2131 et. seq. Implementing regulations are published in the Code of Federal Regulations (CFR), Title 9, Chapter 1, Subchapter A, Parts 1, 2, and 3. 3. Animal Welfare Information Center [Internet]. Pain and distress references: 2007. Available at: 4. Assessing the Health and Welfare of Laboratory Animals [Internet]. Tutorials: 2005. Available 5. Brondani JT, Luna, SPL, Beier, SL, Minto, BW. 2009. Analgesic efficacy of perioperative use of vedaprofen, tramadol or their combination in cats undergoing ovariohysterectomy. J Feline 6. Carbone, Larry, 2007. Pain Management Standards in the Eight Edition of the Guide for the Care and Use of Laboratory Animals. JAALAS 51:(3) 322-328. 7. Carpenter JW, Exotic animal formulary, 3rd edition. 2005. St. Louis (MO): Elsevier. 8. Chappell, MG,Koeller CA, and,Hall, S. I. 2011. Differences in Postsurgical recovery of CF1 Mice after Intraperitoneal Implantation of Radiotelemetry Devices through a Midline or Flank Surgical 9. Committee on Recognition and Alleviation of Pain in Laboratory Animals. 2009. Recognition and Al eviation of Pain in Laboratory Animals. Washington, D.C.: The National Academies Press. 10. Fish RE, Danneman PJ, Brown M, Karas AZ. 2008. Anesthesia and analgesia in laboratory animals, 2nd edition. London (UK): Academic Press. 11. Flecknell PD, Waterman-Pearson A. 2000. Pain management in animals. London: WB Saunders. 12. Flecknell PD, Laboratory animal anesthesia. 1996. 2nd edition. San Diego (CA): Academic Press, 13. Foley, PL, Liang, H, and Crichlow, AR. 2011. Evaluation of a Sustained-Release Formulation of Buprenorphine for Analgesia in Rats. 50:(2) 198-204. 14. Fowler ME, Miller RE. Zoo and wildlife animal medicine, 5th edition. 2005. Philadelphia (PA): WB 15. Gaynor JS, Muir W. 2002. Handbook of veterinary pain management. St. Louis (MO): Mosby. 16. Giamberardino MA, Affaitati G, Lerza R, Vecchiet L. 2000. Pre-emptive analgesia in rats with artificial ureteric calculosis--effects on visceral pain behavior in the post-operative period. Brain 17. Gonzalez MI, Field MJ, Bramwell S, et al. 2000. Ovariohysterectomy in the rat: a model of surgical pain for evaluation of pre-emptive analgesia? Pain 88:79-88. 18. Hawk CT, Leary SL, Morris TH. Formulary for laboratory animals, 3rd edition. 2005. Ames (IA): 19. Humane endpoints for animals used in biomedical research and testing. 2000. ILAR 41:2. 20. Inanoglu, K, Akkurt, BC, Turhanoglu, S, Okuyucu, S, Akoglu, E. 2009. Intravenous keatamine and local buivacine infiltration are effective as part of a multimodal regine for reducing post- tonsillectormy pain. Med Sci Monit. 15(10): CR539-543. 21. IRAC (Interagency Research Animal Committee). 1985. U.S. Government Principles for the Utilization and Care of Vertebrate Animals Used in Testing, Research and Training. Federal Register, May 20, 1985. Washington, D.C.: Office of Science and Technology Policy. 22. Jacobsen, KR, Kalliokoshi, O, Teilmann, AC, Hau, J, and Abelson, SP. 2012. Postsurgical Food and Water Consumption, Fecal Corticosterone Metabolites, and Behavior Assessment as Noninvasive Measure of Pain In Vasectomized BALB/c Mice. JAALAS 51:(1) 69-75. 23. Jensen K, Kehlet H, Lund CM. 2009. Postoperative recovery profile after elective abdominal hysterectomy: a prospective, observational study of a multimodal anaesthetic regime. Eur J 24. Kohn DF, Martin ME, Foley PL, etal. 2007. Guidelines for the assessment and management of pain in rodents and rabbits. (ACLAM position paper, 2006) JAALAS 46:(2) 97-108. 25. Kohn DF, Wixson SK, White WJ, Benson GJ. 1997. Anesthesia and analgesia in laboratory animals. San Diego (CA): Academic Press. 26. Matsumiya, LC, Sorge, RE, Sotocinal, SG, et al. 2012. Using the Mouse Grimace Scale to Reevaluate the Efficacy of Postoperative Analgesics in Laboratory Mice. JAALAS. 51:(1) 42-49. 27. Matthews EA, Dickerson, AH. 2002. A combination of gabapentin and morphine mediates enhanced inhibitory effects on dorsal horn neuronal responses in a rat model of neuropathy. 28. Muir W, Hubell, JA. 2007. Handbook of veterinary anesthesia, 4th edition. St. Louis (MO): Mosby. 29. NIH Policy Manual 3040-2, Animal Care and Use in the Intramural Program. 30. NRC (National Research Council). 2011. Guide for the Care and Use of Laboratory Animals. Washington, D.C.: National Academy Press. 31. Parker, JM< Austin, J, Wilkerson, and Corbone, L. 2011. Effects of Mutimodal Analgesia on the success of Mouse Embryo Transfer Surgery. JAALAS 50:(4) 466-470. 32. PHS (Public Health Service). 1996. Public Health Service Policy on Humane Care and Use of Laboratory Animals. Washington, D.C.: U.S. Department of Health and Human Services, 28 pp. [PL-99-158, Health Research Extension Act, 1985] 33. Plumb DC. 2008. Veterinary drug handbook, 6th edition. Wiley-Blackwell Publishing. 34. Recognition and al eviation of pain and distress in laboratory animals. 1992. ILAR. Washington 35. Robertson SA. 2005. Managing pain in feline patients. Vet Clin Smal Anim. 35: 129-146. 36. Roughan JV, Flecknel PA. 2000. Effects of surgery and analgesic administration on spontaneous behavior in singly housed rats. Res in Vet Sci 69: 283-288. 37. Roughan JV, Flecknell PA. 2001. Behavioural effects of laporotomy and analgesic effects of ketoprofen and carprofen in rats. Pain 90: 65-74. 38. Roughan JV, Flecknell PA. 2003. Evaluation of a short duration behaviour-based post-operative pain scoring system in rats. Eur J of Pain 7:397-406. 39. Smith G, Covino BG. 1985. Acute Pain. London: Butterworths. 40. Suzuki, M. 2009. Role of N-methyl-D-aspartate receptor antagonists in the postoperative pain management. Curr Opin Anaesthsiol. 22: 618-622. TABLE 1: POTENTIAL SIGNS ASSOCIATED WITH PAIN OR DISTRESS IN RATS, MICE AND RABBITS
Mice Rats Rabbits
Unkempt Appearance (Erected, Matted, or Dul Haircoat) Abnormal Posture/Positioning (e.g., Head-pressing, Hunched Back) Tearing (including Porphyria), Lack of Blinking Reflex Redness or Swelling Around Surgical Site TABLE 2: POST PROCEDURAL PAIN POTENTIAL a,b
Minimal to Mild c
Mild to Moderate d
Moderate to Severe e
Subcutaneous transponder placement C-section a Table adapted from “Guidelines for the Assessment and Management of Pain in Rodents and Rabbits”. 2006, American College of Laboratory Animal Medicine b The analgesia and monitoring required may vary due to a number of factors; such as the invasiveness of the procedure, degree of tissue trauma, surgical time, skil of the surgeon, and the tissues or organs involved. c Post procedural pain relief for minimal to mild pain may be adequately addressed with preemptive analgesia, tissue infiltration with a long acting local anesthetic, and a single dose of a long acting NSAID or mixed opioid d Post procedural pain relief for mild to moderate pain may be adequately addressed with tissue infiltration with a long acting local anesthetic combined with one or more doses of a long acting NSAID and/or an opioid or other agent in addition to pre-emptive analgesic administration. e Post procedural pain relief for moderate to severe pain should encompass multimodal analgesia (e.g. combining a pure opioid agonist with a NSAID, tissue infiltration with a long acting local anesthetic, etc.)



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