Hostname: page-component-78c5997874-94fs2 Total loading time: 0 Render date: 2024-11-19T10:33:49.118Z Has data issue: false hasContentIssue false

Euthanasia methods, corticosterone and haematocrit levels in Xenopus laevis: evidence for differences in stress?

Published online by Cambridge University Press:  01 January 2023

GA Archard*
Affiliation:
406 Forest Resources Building, School of Forest Resources, The Pennsylvania State University, State College, PA 16802, USA School of Biological Sciences, University Bristol, Woodland Road, Bristol BS8 1UG, UK
AR Goldsmith
Affiliation:
School of Biological Sciences, University Bristol, Woodland Road, Bristol BS8 1UG, UK
*
* Contact for correspondence and requests for reprints: [email protected]
Rights & Permissions [Opens in a new window]

Abstract

Core share and HTML view are not available for this content. However, as you have access to this content, a full PDF is available via the ‘Save PDF’ action button.

Amphibians, like other vertebrates, respond to acute stressors by releasing glucocorticoid steroid hormones that mediate physiological and behavioural responses to stress. Measurement of stress hormones provides a potential means to improve the welfare of laboratory animals. For example, manipulations of laboratory housing and procedures combined with measurement of glucocorticoids may identify which conditions are more stressful to animals. This is important because there is very little experimental evidence to guide best practice for welfare in amphibians and other lower vertebrates. We investigated the effect of different methods of euthanasia on the circulating plasma corticosterone levels in the African clawed frog (Xenopus laevis), a model amphibian organism that is frequently used in laboratories. In particular, we investigated the effect of different concentrations and pH of the anaesthetic tricaine methanesulphonate (MS-222). Low concentration and unbuffered (low pH) solutions of MS-222 caused elevated corticosterone levels, but only after the effect of MS-222 treatment on blood fluid volume had been taken into account. The level of disturbance that animals experienced also affected corticosterone levels. Thus, our data suggest that to minimise stress to X. laevis, animals should be euthanised after minimal disturbance and in a 3 g L−1 MS-222 solution, buffered to pH 7. The potential for the improvement of amphibian welfare using corticosterone measures as a tool is discussed.

Type
Research Article
Copyright
© 2010 Universities Federation for Animal Welfare

References

AVMA 2007 AVMA Guidelines on Euthanasia. American Veterinary Medical Association: Schaumberg, IL, USA. Available at: http://www.avma.org/issues/animal welfare/euthanasia.pdfGoogle Scholar
Barton, BA and Peter, RE 1982 Plasma cortisol stress response in fingerling rainbow trout, Salmo gairdneri Richardson, to various transport conditions, anaesthesia, and cold shock. Journal of Fish Biology 20: 3951Google Scholar
Boutilier, RG and Shelton, G 1986 Respiratory properties of blood from voluntary and forcibly submerged Xenopus laevis. Journal of Experimental Biology 121: 285300CrossRefGoogle Scholar
Broom, DM and Johnson, KG 2000 Stress and Animal Welfare, Second Edition. Kluwer Academic Publishers: The NetherlandsGoogle Scholar
Crespi, EJ, Vaudry, H and Denver, RJ 2004 Roles of corticotrophin-releasing factor, neuropeptide Y and corticosterone in the regulation of food intake in Xenopus laevis. Journal of Neuroendocrinology 16: 279288CrossRefGoogle ScholarPubMed
Downes, H 1995 Tricaine anaesthesia in amphibia: a review. Bulletin of the Association of Reptile and Amphibian Veterinarians 5: 1116CrossRefGoogle Scholar
Fox, HE, White, SA, Kao, MHF and Fernald, RD 1997 Stress and dominance in a social fish. Journal of Neuroscience 17: 64636469CrossRefGoogle Scholar
Frangioni, G, Berti, R and Borgioli, G 1997 Hepatic respiratory compensation and haematological changes in the cave cyprinid, Phreatichthys andruzzii. Journal of Comparative Physiology 167B: 461467CrossRefGoogle Scholar
Guardabassi, A, Muccioli, G, Andreoletti, GE, Pattono, P and Usai, P 1991 Prolactin and interrenal hormone balance in Xenopus laevis specimens adapted to brackish water. Accademia Delle Scienze di Torino 125: 5569Google Scholar
Guardabassi, A, Muccioli, G, Andreoletti, GE, Pattono, P and Usai, P 1993 Prolactin and interrenal hormone balance in adult specimens of Xenopus laevis exposed to hyperosmotic stress for up to one week. Journal of Experimental Zoology 265: 515521CrossRefGoogle ScholarPubMed
Gurdon, JB 1996 Introductory comments: Xenopus as a laboratory animal. In: Tinsley, RC and Kobel, HR (eds) The Biology of Xenopus pp 36. Clarendon Press: Oxford, UKGoogle Scholar
Gurdon, JB and Hopwood, N 2000 The introduction of Xenopus laevis into developmental biology: Of empire, pregnancy testing and ribosomal genes. International Journal of Developmental Biology 44: 4350Google ScholarPubMed
Homan, RN, Recosin, JV, Rodrigues, DM, Reed, JM, Windmiller, BS and Romero, LM 2003 Impacts of varying habitat quality on the physiological stress of spotted salamanders (Ambystoma maculatum). Animal Conservation 6: 1118CrossRefGoogle Scholar
Home Office 1987 Code of Practice for the Humane Killing of Animals under Schedule 1 to the Animals (Scientific Procedures) Act 1986. Available at: http://scienceandresearch.homeoffice.gov.uk/animal-research/publications-and-reference/publications/code-of-practice/code-of-practice-housing-care/humane_killing.pdf?view+BinaryGoogle Scholar
Hopkins, WA, Medonça, MT and Congdon, JD 1997 Increased circulating levels of testosterone and corticosterone in Southern toads, Bufo terrestris, exposed to coal combustion waste. General and Comparative Endocrinology 108: 237246CrossRefGoogle ScholarPubMed
King, WV, Hooper, B, Hillsgrove, S, Benton, C and Berlinsky, DL 2005 The use of clove oil, metomidate, tricaine methanesulphonate and 2-phenoxyethanol for inducing anaesthesia and their effects on the cortisol stress response in Black sea bass (Centropristis striata L). Aquaculture Research 36: 14421449CrossRefGoogle Scholar
Lowe, J 2004 Rates of tricaine methanesulfonate (MS-222) anaesthetization in relation to pH and concentration in five terrestrial salamanders. Herpetology Review 35: 352354Google Scholar
MacAvoy, SE and Zaepfel, RC 1997 Effects of tricaine methanesulfonate (MS-222) on hematocrit: First field measurements on blacknose dace. Transactions of the American Fisheries Society 126: 5005032.3.CO;2>CrossRefGoogle Scholar
Maddocks, SA, Goldsmith, AR and Cuthill, IC 2001 The influence of flicker rate on plasma corticosterone levels of European starlings, Sturnus vulgaris. General and Comparative Endocrinology 124: 315320CrossRefGoogle ScholarPubMed
Molinero, A and Gonzalez, J 1995 Comparative effects of MS-222 and 2-Phenoxyethanol on gilthead sea bream (Sparus aurata L) during confinement. Comparative Biochemistry and Physiology 111A: 405414CrossRefGoogle Scholar
Morgan, KN and Tromborg, CT 2007 Sources of stress in captivity. Applied Animal Behaviour Science 102: 262302CrossRefGoogle Scholar
Ohr, EA 1976a Tricaine methanesulphonate - I. pH and its effects on anesthetic potency. Comparative Biochemistry and Physiology 54C: 1317Google Scholar
Ohr, EA 1976b Tricaine methanesulphonate - II. Effects on transport of NaCl and H2O. Comparative Biochemistry and Physiology 54C: 1922Google Scholar
Read, BT 2005 Guidance on the Housing and Care of the African Clawed Frog Xenopus laevis. RSPCA: Horsham, UKGoogle Scholar
Reilly, JS 2001 Euthanasia of Animals used for Scientific Purposes. Australian and New Zealand Council for the Care of Animals in Research and Teaching: Adelaide, AustraliaGoogle Scholar
Romero, LM 2004 Physiological stress in ecology: lessons from biomedical research. Trends in Ecology and Evolution 19: 249255CrossRefGoogle ScholarPubMed
Scott, AP and Ellis, T 2007 Measurement of fish steroids in water: a review. General and Comparative Endocrinology 153: 392400Google ScholarPubMed
Smit, GL and Hattingh, J 1979 Anaesthetic potency of MS 222 and neutralized MS 222 as studied in three freshwater fish species. Comparative Biochemistry and Physiology 62C: 237241Google Scholar
Wingfield, JC, Vleck, CM and Moore, MC 1992 Seasonal changes in the adrenocortical response to stress in birds of the Sonoran Desert. Journal of Experimental Zoology 264: 419428CrossRefGoogle ScholarPubMed
Witters, HE, Vanpuymbroeck, S, Vandensande, I and Vanderborght, OLJ 1990 Hematological disturbances and osmotic shifts in rainbow-trout, Oncorhynchus mykiss (Walbaum) under acid and aluminum exposure. Journal of Comparative Physiology 160B: 563571Google Scholar
Yao, M, Westphal, NJ and Denver, RJ 2004 Distribution and acute stressor-induced activation of corticotrophin-releasing hormone neurons in the central nervous system of Xenopus laevis. Journal of Neuroendocrinology 16: 880893CrossRefGoogle ScholarPubMed
Ziegler, DR and Herman, JP 2002 Neurocircuitry of stress integration: anatomical pathways regulating the hypothalamo-pituitary-adrenocortical axis of the rat. Integrative and Comparative Biology 42: 541551CrossRefGoogle ScholarPubMed