Laboratory animals anesthesia recommendations for biomedical research purposes

Anesthesia is widely used in experimental biomedical research to anesthetize and immobilize laboratory animals during surgical interventions and experimental procedures. In contrast to clinical and veterinary anesthesiology, an important aspect of anesthesia in laboratory animals is to minimize the effect of anesthetics on the results of the study. On November 19, 2022, the IV Scientific and Practical Conference “Experimental Surgery, Anesthesiology and Reanimatology of Laboratory Animals” was held in Moscow, where the community of specialists in the field of laboratory animal science, experimental surgery, pathophysiology, veterinary medicine, clinical and veterinary anesthesiology discussed various aspects of anesthesia in laboratory animals. This review is based on the conference proceedings and is devoted to general issues of animal anesthesia, its technological aspects (methods, equipment and consumables), consideration of advantages and disadvantages of the most commonly used inhalation and injection anesthetics.

The advantages and schemes of modern combined anesthesia are substantiated, in particular, the need for adequate intraoperative analgesia and the possibility of regional anesthesia. The use of obsolete and non-recommended drugs is also discussed. The review and recommendations have a practical orientation and are intended for biomedical researchers who use laboratory animals in basic, translational and regulated preclinical studies.

1. Fish RE, Brown MJ, Danneman PJ, Karas AZ (eds.). Anesthesia and Analgesia in Laboratory Animals, 2nd ed. CA: Academic Press; 2008: 656.

2. Flecknell P. Laboratory Animal Anaesthesia, 4th ed. Academic Press; 2016: 300.

3. Tranquilli WJ, Thurmon JC, Grimm KA, editors. Lumb and Jones’ veterinary anesthesia and analgesia. Hoboken, NJ: John Wiley & Sons; 2013.

4. Karkischenko NN, Grachev SV. (red) Guidance on Laboratory Animals and Alternative Models in Biomedical Research. M.: Profli’. 2010. In Russian.

5. Smith AJ. Guidelines for planning and conducting high-quality research and testing on animals. Lab Anim Res. 2020; 36:21. DOI: 10.1186/s42826-020-00054-0.

6. Percie du Sert N, Hurst V, Ahluwalia A, et al. The ARRIVE guidelines 2.0: Updated guidelines for reporting animal research. PLoS Biol. 2020; 18(7): e3000410. DOI: 10.1371/journal.pbio.3000410.

7. Uhlig C, Krause H, Koch T, et al. Anesthesia and Monitoring in Small Laboratory Mammals Used in Anesthesiology, Respiratory and Critical Care Research: A Systematic Review on the Current Reporting in Top-10 Impact Factor Ranked Journals. PLoS One. 2015; 25;10:8: e0134205. DOI: 10.1371/journal.pone.0134205.

8. Fergusson DA, Avey MT, Barron CC, et al. Canadian Perioperative Anesthesia Clinical Trials Group. Reporting preclinical anesthesia study (REPEAT): Evaluating the quality of reporting in the preclinical anesthesiology literature. PLoS One. 2019; 14:5: e0215221. DOI: 10.1371/journal.pone.0215221.

9. Bunyatyan AA, Ryabov GA, Manevich AZ. Anesthesiology and Rehanimatology. M.: Meditsina. 1984. In Russian.

10. Meyer RE, Fish RE. Pharmacology of Injectable Anesthetics, Sedatives, and Tranquilizers. In: Fish RE, Brown MJ, Danneman PJ, Karas AZ (eds.). Anesthesia and Analgesia in Laboratory Animals, 2nd ed. CA: Academic Press. 2008: 27–82. DOI: 10.1016/B978-012373898-1.50006-1.

11. Davies NJH, Cashman JN. (eds.) Lee’s Synopsis of Anaesthesia, 13th ed. Elsevier Health Sciences. 2005: 948. DOI: 10.1016/j.cacc.2006.02.006.

12. Brunson DB. Pharmacology of Inhalation Anesthetics. In: Fish RE, Brown MJ, Danneman PJ, Karas AZ (eds.). Anesthesia and Analgesia in Laboratory Animals, 2nd ed. CA: Academic Press. 2008: 83–95.

13. Navarro KL, Huss M, Smith JC, et al. Mouse Anesthesia: The Art and Science. ILAR J. 2021; 62:1–2; 238–273. DOI: 10.1093/ilar/ilab016.

14. Bigiarelli K, Schepers LE, Soepriatna AH, et al. Use of an Integrated Low-Flow Anesthetic Vaporizer, Ventilator, and Physiological Monitoring System for Rodents. J Vis Exp. 2020; 161: 10.3791/61311. DOI: 10.3791/61311.

15. Ambrisko TD, Klide AM. Evaluation of isoflurane and sevoflurane vaporizers over a wide range of oxygen flow rates. Am J Vet Res. 2006; 67:6; 936–940. DOI: 10.2460/ajvr.67.6.936.

16. Adelsperger AR, Bigiarelli-Nogas KJ, Toore I, et al. Use of a Low-flow Digital Anesthesia System for Mice and Rats. J Vis Exp. 2016; 7:115; 54436. DOI: 10.3791/54436.

17. Damen FW, Adelsperger AR, Wilson KE, et al. Comparison of traditional and integrated digital anesthetic vaporizers. JAALAS. 2015; 54:6; 756–762.

18. Lapin KN, Ryzhkov IA, Maltseva VA, et al. Catheterization of vessels of small laboratory animals during biomedical research: technological aspects of the method (overview). Siberian Medicine Bulletin. 2021; 20:3; 168–181. In Russian. DOI: 10.20538/1682-0363-2021-3-168-181.

19. Swindle M. Second edition: Swine in the laboratory: Surgery, anesthesia, imaging, and experimental techniques, 2007. DOI: 10.1201/9781420009156.

20. McCormick B. (ed.) Basic course of anesthesiologist: textbook, electronic version/translation from English ed. E. V. Nedashkovsky, V. V. Kuzkov. Arkhangelsk: Northern State Medical University. 2010: 238. In Russian.

21. Loer SA, Scheeren TWL, Tarnow J. Desflurane Inhibits Hypoxic Pulmonary Vasoconstriction in Isolated Rabbit Lungs. Anesthesiology. 1995; 83; 552–556. DOI: 10.1097/00000542-199509000-00014.

22. Hedenqvist P, Roughan JV, Antunes L, et al. Induction of anaesthesia with desflurane and isoflurane in the rabbit. Lab Anim. 2001; 35:2; 172–9. DOI: 10.1258/0023677011911561.

23. Gaertner DJ, Hallman TM, Hankenson FC, et al. Anesthesia and Analgesia for Laboratory Rodents. In: Fish RE, Brown MJ, Danneman PJ, Karas AZ (eds.). Anesthesia and Analgesia in Laboratory Animals, 2nd ed. CA: Academic Press. 2008: 239–297.

24. Royse CF, Liew DFL, Wright CE, et al. Persistent Depression of Contractility and Vasodilation with Propofol but Not with Sevoflurane or Desflurane in Rabbits. Anesthesiology. 2008; 108:1; 87–93. DOI: 10.1097/01.anes.0000296077.32685.26.

25. Kilicaslan A, Belviranli M, Okudan N, et al. Single and repeated sevoflurane or desflurane exposure does not impair spatial memory performance of young adult mice. Fundam Clin Pharmacol. 2013; 27:6; 641–9. DOI: 10.1111/fcp.12027.

26. Niikura R, Miyazaki T, Takase K, et al. Assessments of prolonged effects of desflurane and sevoflurane on motor learning deficits in aged AppNL-G-F/ NL-G-F mice. Mol Brain. 2022; 7; 15:1; 32. DOI: 10.1186/s13041-022-00910-1.

27. Hedley J. (ed.) BSAVA Small Animal Formulary 10th ed. Part B: Exotic Pets. British small animal veterinary association; 2020: 348.

28. Protsak ES, Borshchev YuYu, Galagudza MM. The role of estimating basic hemodynamic parameters in modern experimental practice. Regional circulation and microcirculation. 2023; 22:1; 103–109. In Russian. DOI: 10.24884/1682-6655-2023-22-1-103-109.

29. Wenger S. Anesthesia and Analgesia in Rabbits and Rodents. Journal of Exotic Pet Medicine. 2012; 21:1; 7–16. DOI: 10.1053/j.jepm.2011.11.010.

30. Mikołajczyk A. Safe and effective anaesthesiological protocols in domestic pig. Ann. Warsaw Univ. Life Sci. – SGGW, Anim. Sci. 2016; 55:2; 219–227.

31. Tendillo FJ, Mascías A, Santos M, et al. Cardiopulmonary and analgesic effects of xylazine, detomidine, medetomidine, and the antagonist atipamezole in isoflurane-anesthetized swine. Lab Anim Sci. 1996; 46:2; 215–9.

32. Turner PV, Albassam MA. Susceptibility of rats to corneal lesions after injectable anesthesia. Comp Med. 2005; 55:2; 175–82.

33. Doerning BJ, Brammer DW, Chrisp CE, et al. Nephrotoxicity of tiletamine in New Zealand white rabbits. Lab Anim Sci. 1992; 42:3; 267–9.

34. Hull RM. Guideline Limit Volumes for Dosing Animals in The Preclinical Stage of Safety Evaluation. Human & Experimental Toxicology. 1995; 14:3; 305–307. DOI: 10.1177/096032719501400312.

35. Diehl KH, Hull R, Morton D, et al. A Good Practice Guide to the Administration of Substances and Removal of Blood Including Routes and Volumes. Journal of Applied Toxicology. 2001; 21:1; 15–23. DOI: 10.1002/jat.727.

36. Morton DB, Jenning M, Buckwell A, et al. Refining Procedures for the Administration of Substances. Report of the BVAAWF/FRAME/RSPCA/UFAW Joint Working Group on Refinement. Laboratory Animals. 2001; 35:1; 1–41. DOI: 10.1258/0023677011911345.

37. Ahmadi-Noorbakhsh S, Farajli Abbasi M, Ghasemi M, et al. Anesthesia and analgesia for common research models of adult mice. Lab Anim Res. 2022; 13:38:1; 40. DOI: 10.1186/s42826-022-00150-3.

38. Corletto F. Multimodal and balanced analgesia. Vet Res Commun. 2007; 1; 59–63. DOI: 10.1007/s11259-007-0085-5.

39. Ovechkin AM, Bayalieva AZ, Yezhevskaya AA, et al. Postoperative pain relief. Clinical guidelines. Bulletin of Intensive Care named after A. I. Saltanov. 2019; 4; 9–33. In Russian. DOI: 10.21320/1818-474X-2019-4-9-33.

40. Burton M, Conway R, Mishkin N, et al. Pharmacokinetics of gabapentin after single, oral administration in domestic rabbits (Oryctolagus cuniculus). Journal of Exotic Pet Medicine. 2023; 45:5; 1–5. DOI: 10.1053/J.JEPM.2023.02.001.

41. d’Ovidio D, Adami C. Locoregional Anesthesia in Exotic Pets. Vet Clin North Am Exot Anim Pract. 2019; 22:2; 301–314. DOI: 10.1016/j.cvex.2019.01.007.

42. Lerche P, Aarnes T, Covey-Crump G, et al. Handbook of Small Animal Regional Anesthesia and Analgesia Techniques. 2016. DOI:10.1002/9781119159490.

43. Skarda RT. Local and regional anesthesia in ruminants and swine. Vet Clin North Am Food Anim Pract. 1996; 12:3; 579–626. DOI: 10.1016/s0749-0720(15)30390-x.

44. Ouchi K, Sekine J, Koga Y, et al. Establishment of an animal model of sedation using epidural anesthesia that uses the tail-flick test for evaluating local anesthetic effects in rats. Exp Anim. 2013; 62:2; 137–44. DOI: 10.1538/expanim.62.137.

45. Adolphs J, Schmidt DK, Mousa SA, et al. Thoracic epidural anesthesia attenuates hemorrhage-induced impairment of intestinal perfusion in rats. Anesthesiology. 2003; 99:3; 685–92. DOI: 10.1097/00000542-200309000-00025.

46. Rahman MM, Lee JY, Kim YH, et al. Epidural and Intrathecal Drug Delivery in Rats and Mice for Experimental Research: Fundamental Concepts, Techniques, Precaution, and Application. Biomedicines 2023; 10:11:5; 1413. DOI: 10.3390/biomedicines11051413.

47. Kim NH, Lee SH, Lee SJ. Percutaneous transforaminal epidural injection method in an experimental rat: minimally invasive drug delivery method to spinal epidural space. Ann Rehabil Med. 2012; 36:5; 640–647. DOI: 10.5535/arm.2012.36.5.640.

48. Yashpal K, Katz J, Coderre TJ. Effects of Preemptive or Postinjury Intrathecal Local Anesthesia on Persistent Nociceptive Responses in Rats: Confounding Influences of Peripheral Inflammation and the General Anesthetic Regimen. Anesthesiology. 1996; 84:5; 1119–1128. DOI: 10.1097/00000542-199605000-00014.

49. Yahalom B, Athiraman U, Soriano SG, et al. Spinal anesthesia in infant rats: development of a model and assessment of neurologic outcomes. Anesthesiology. 2011; 114:6; 1325–35. DOI: 10.1097/ALN.0b013e31821b5729.

50. Carbone E, Lindstrom K, Diep S, et al. Duration of action of sustained-release buprenorphine in 2 strains of mice. JALAAS. 2012; 51:6; 815–819.

51. Mayer J, Mans C. Rodents. In: Carpenter J, Marion C, eds. Exotic Animal Formulary. 5thed. St. Louis, MO: Elsevier. 2018; 459–493.

52. Foley P, Kendall L, Turner P. Clinical management of pain in rodents. Comp Med. 2019; 69:6; 468–489. DOI: 10.30802/AALAS-CM-19-000048.

53. Morrisey J, Carpenter J. Ferrets, Rabbits, and Rodents: Clinical Medicine and Surgery. 3rd ed. St. Louis, MO: Saunders/Elservier; 2012.

54. Plumb D. Plumb’s Veterinary Drug Handbook. Blackwell Pub: Electronic-App; 2019.

55. Nishiyori M, Ueda H. Prolonged gabapentin analgesia in an experimental mouse model of fibromyalgia. Mol Pain. 2008; 6:4; 52. DOI: 10.1186/1744-8069-4-52.

56. Tubbs JT, Kissling GE, Travlos GS, et al. Effects of buprenorphine, meloxicam, and flunixin meglumine as postoperative analgesia in mice. J Am Assoc Lab Anim Sci. 2011; 50:2; 185–91.

57. Kang SC, Jampachairsri K, Seymour TL, et al. Use of liposomal bupivacaine for postoperative analgesia in an incisional pain model in rats (Rattus norvegicus). JALAAS. 2017; 56:1; 63–68.

58. Durst MS, Arras M, Palme R, et al. Lidocaine and bupivacaine as part of multimodal pain management in a C57BL/6J laparotomy mouse model. Sci Rep. 2021; 25:11:1; 10918. DOI: 10.1038/s41598-021-90331-2.

59. Van Pelt LF. Ketamine and xylazine for surgical anesthesia in rats. J Am Vet Med Assoc. 1977; 1:171:9; 842–4.

60. Grint NJ, Murison PJ. A comparison of ketamine-midazolam and ketamine-medetomidine combinations for induction of anaesthesia in rabbits. Vet Anaesth Analg. 2008; 35:2; 113–21. DOI: 10.1111/j.1467-2995.2007.00362.x.

61. Cagle LA, Franzi LM, Epstein SE, et al. Injectable Anesthesia for Mice: Combined Effects of Dexmedetomidine, Tiletamine-Zolazepam, and Butorphanol. Anesthesiol Res Pract. 2017; 2017:9161040. DOI: 10.1155/2017/9161040.

62. Vovk AN, Karantysh G, Kosenko PO, et al. Principles of therapy and care of laboratory animals after chronic administration into Xylasine-ZoletylВ® Anesthesia. International Journal of Veterinary Science and Research. 2020; 114–117. DOI: 10.17352/ijvsr.000062.

63. Limprasutr V, Sharp P, Jampachaisri K, et al. Tiletamine/zolazepam and dexmedetomidine with tramadol provide effective general anesthesia in rats. Animal Model Exp Med. 2021; 2:4:1; 40–46. DOI: 10.1002/ame2.12143.

64. Baxter MG, Murphy KL, Taylor PM, et al. Chloral hydrate is not acceptable for anesthesia or euthanasia of small animals. Anesthesiology. 2009; 111:1; 209. DOI: 10.1097/ALN.0b013e3181a8617e.

65. Silverman J, Muir WW. 3rd. A review of laboratory animal anesthesia with chloral hydrate and chloralose. Lab Anim Sci. 1993; 43:3; 210–6.

66. Bilan DS, Kelmanson IV, Belousov VV. Influence of the type of anesthesia and the conditions of brain tissue dyeing with 2.3 dye, 5-triphenyltetrazolium chloride (TTC) on the assessment of ischemic brain damage in rats in the early stages of pathogenesis. Bulletin of the Russian State Medical University. 2017; 6; 67–74. In Russian. DOI: 10.24075/vrgmu.2017-06-11.

67. Silachev DN, Usatikova EA, Pevzner IB, et al. Effect of Anesthetics on Efficiency of Remote Ischemic Preconditioning. Biochemistry (Mosc). 2017; 82:9; 1006–1016. DOI: 10.1134/S0006297917090036.