Behavioural Effects of the Commonly Used Fish Anaesthetic Tricaine Methanesulfonate (MS-222) on Zebrafish (Danio rerio) and Its Relevance for the Acetic Acid Pain Test

Abstract

The pros and cons of using anaesthesia when handling fish in connection with experiments are debated. A widely adopted practice is to wait thirty minutes after anaesthesia before behavioural observations are initiated, but information about immediate effects of a treatment is then lost. This is pertinent for responses to acute stressors, such as acid injection in the acetic acid pain test. However, omission of anaesthetics in order to obtain data on immediate responses will compromise the welfare of fish and contribute to experimental noise due to stress. We therefore tested the effect of tricaine methanesulfonate on the behaviour of zebrafish. We predicted that tricaine (MS 222) would decrease swimming velocity and that the control fish would show an increased level of anxiety- and stress-related behaviours compared to the tricaine group. Following acclimatization to the test tank, baseline behaviour was recorded before immersion in either tricaine (168 mg l−1, treatment group, N = 8) or tank water (control group, N = 7). Latencies to lose equilibrium and to lose response to touch were registered. The fish was then returned to the test tank, and the latency to regain equilibrium was registered in anaesthetized fish. When equilibrium was regained, and at five, thirty and sixty minutes after the fish had been returned to the test tank, behaviour was recorded. The tricaine fish showed the following responses (mean ± sd): latency to lose equilibrium 22.6 s±3.9; latency to lose response to touch 101.9 s±26.8; latency to regain equilibrium 92.0 s±54.4. Contrary to our predictions, neither treatment caused a change in any of the behaviours registered. This indicates that tricaine has no effect on several commonly used behavioural parameters, and that it may be unnecessary to postpone behavioural observations to 30 min after anaesthesia.

Reproductive success of a marine teleost was correlated with proactive and reactive stress‐coping styles

“…The second test, confinement test, consisted of evaluating for 300 s the reaction of fish to confinement (the fish was placed in a small tank (56 cm length × 36 cm width × 30 cm depth) with a constant flow through of oxygenated water) by evaluating two variables: the latency to the first activity time (s) of fish in confinement (ConLat) and the total activity time (s) in confinement (ConAct) ( Lepomis macrochirus Rafinesque 1819 (Wilson & Godin, ); Cyprinus carpio L. 1758 (Huntingford et al ., ); G. aculeatus (King et al ., ); S. aurata (Herrera et al ., ); Dicentrarchus labrax (L. 1758) (Ferrari et al ., ); S. senegalensis (Ibarra‐Zatarain et al ., ). The third trial, anaesthesia test, consisted of evaluating the time required (s) to induce 5 different anaesthesia levels (Table ) ( Ictalurus punctatus (Rafinesque 1818) (Welker et al ., ); Danio rerio (Hamilton 1822) (Nordgreen et al ., ). The anaesthesia agent used was buffered 100 mg l −1 MS‐222 (Norambuena et al ., ).…”

“…The first test, restraining test, consisted of netting fish individually in the water and three behavioural variables were evaluated for 120 s: (Welker et al, 2007); Danio rerio (Hamilton 1822) (Nordgreen et al, 2014). The anaesthesia agent used was buffered 100 mg l −1 MS-222 (Norambuena et al, 2011).…”

Use of MS-222 (tricaine methanesulfonate) and propofol (2,6-diisopropylphenol) as anaesthetics for the tetraAstyanax altiparanae(Teleostei, Characidae)

“…MS-222 is supplied as a white crystalline powder, and its main properties include high solubility in water and rapid induction and full recovery of animals subjected to anaesthesia (Hseu, Yeh, Chu & Ting 1998;Roberts 2009;Readman, Owen, Murrell & Knowles 2013). MS-222 has been used on freshwater and marine fish (Lemm 1993;Hseu et al 1998;Roubach, Gomes & Val 2001;Sladky, Swanson, Stoskopf, Loomis & Lewbart 2001;Welker, Lim & Yildirim-Aksoy 2007;Ross & Ross 2008;Zahl, Kiessling, Samuelsen & Olsen 2010;Gholipour, Mirzargar, Soltani, Ahmadi, Abrishamifar, Bahonar & Yousefi 2011;Stockman, Weber, Kass, Pascoes & Paul-Murphy 2012;Ribeiro, de Melo, do Espirito Santo, de Souza e Silva, Santos & Luz 2013;Gressler, Riffel, Parodi, Saccol, Koakoski, DaCosta, Pavanato, Heinzmann, Caron, Schmidt, Llesuy, Barcellos & Baldisserotto 2014;Nordgreen, Tahamtani, Janczak & Horsberg 2014). Moreover, it is the only anaesthetic approved for use by the US Food and Drug Administration (FDA 1997;Carter, Woodley & Brown 2011;Delbon & Ranzani-Paiva 2012).…”

Hematological effects of etomidate and tricaine in common carp

“…Longer recovery of fish from etomidate anesthesia compared to that obtained with tricaine was reported by Kristan et al (9). Nordgreen et al (25) observed that tricaine had no effect on fish swimming behavior, which returned to a normal pattern immediately after recovery from anesthesia. Hematological results obtained 1 h after treatment revealed that both anesthetics affected the values of blood parameters very little in common carp juveniles.…”