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However, species-specific reactions and side-effects have arisen from the use of the most common anaesthetics (MS-222 and eugenol). This study investigated the potential of menthol, a naturally derived and environmentally friendly compound, as an alternative immersion anaesthetic for adult zebrafish (\n                    <jats:italic>Danio rerio<\/jats:italic>\n                    ) through concentration-response assays and the evaluation of cardiorespiratory, locomotion and aversive-related effects. Concentrations below 25\u00a0mg\/L did not induce anaesthetic states while concentrations of 50 and 75\u00a0mg\/mL were considered the most effective, as they successfully induced anaesthesia with induction and recovery times below 400\u00a0s. These values were comparable to those observed with MS-222. In addition, no mortality was observed at these concentrations despite the transient depression of the cardio-respiratory system. The video tracking analyses showed no significant behavioural changes for the lowest concentrations while relatively narrow behavioural safety margins were noticed for the highest concentrations (&gt;\u2009150\u00a0mg\/L). In addition, concentrations above 75\u00a0mg\/L induced aversive behaviours which do not support their use for fish anaesthesia. Based on these findings, a short-exposure (10-min) was conducted using menthol concentration of 50\u00a0mg\/L, resulting in cortisol values equal to those of the control group. Overall, this study shows that a menthol concentration of 50\u00a0mg\/L can be used as an alternative and more sustainable anaesthetic for zebrafish while higher concentrations may compromise fish welfare.\n                  <\/jats:p>","DOI":"10.1007\/s11259-025-10860-3","type":"journal-article","created":{"date-parts":[[2025,8,26]],"date-time":"2025-08-26T04:01:39Z","timestamp":1756180899000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Menthol as a sustainable alternative anaesthetic for adult zebrafish (Danio rerio)"],"prefix":"10.1007","volume":"49","author":[{"given":"Lu\u00eds","family":"F\u00e9lix","sequence":"first","affiliation":[]},{"given":"Sandra M.","family":"Monteiro","sequence":"additional","affiliation":[]},{"given":"Carlos","family":"Ven\u00e2ncio","sequence":"additional","affiliation":[]}],"member":"297","published-online":{"date-parts":[[2025,8,26]]},"reference":[{"issue":"12","key":"10860_CR1","doi-asserted-by":"publisher","first-page":"895","DOI":"10.1007\/s00359-016-1129-5","volume":"202","author":"MS Abreu","year":"2016","unstructured":"Abreu MS, Giacomini AC, Gusso D, Koakoski G, Oliveira TA, Marqueze A, Barreto RE, Barcellos LJ (2016) Behavioral responses of zebrafish depend on the type of threatening chemical cues. 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