{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,19]],"date-time":"2026-06-19T04:25:44Z","timestamp":1781843144439,"version":"3.54.5"},"reference-count":13,"publisher":"Oxford University Press (OUP)","issue":"2","license":[{"start":{"date-parts":[[2025,12,4]],"date-time":"2025-12-04T00:00:00Z","timestamp":1764806400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/pages\/standard-publication-reuse-rights"}],"funder":[{"name":"Environment Research and Technology Development Fund","award":["JPMEERF20235004"],"award-info":[{"award-number":["JPMEERF20235004"]}]},{"DOI":"10.13039\/100014423","name":"Environmental Restoration and Conservation Agency of Japan","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100014423","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002241","name":"Japan Science and Technology Agency","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100002241","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100025019","name":"Support for Pioneering Research Initiated by the Next Generation","doi-asserted-by":"publisher","award":["JPMJSP2124"],"award-info":[{"award-number":["JPMJSP2124"]}],"id":[{"id":"10.13039\/501100025019","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,2,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>In acute fish toxicity tests, mortality has traditionally served as the primary endpoint. However, in accordance with the \u201c3Rs\u201d principle\u2014replacement, reduction, and refinement\u2014there is a growing need to minimize the suffering and pain experienced by test fish. In this study we aimed to establish a behavioral framework for identifying the moribund state in zebrafish (Danio rerio), providing a humane and ethically refined alternative endpoint. Continual observation of zebrafish exposed to 10 representative chemicals allowed the documentation of 12 clinical signs, with the severity of the signs evaluated by the death\/clinical-sign ratio, which represents the proportion of fish exhibiting a given sign that subsequently died. The signs \u201cimmobility,\u201d \u201cimmobility at surface,\u201d and \u201clethargy\u201d emerged as strong predictors of imminent death, each exhibiting a death\/clinical-sign ratio of 1.0 across all tested chemicals and concentrations, indicating that all fish exhibiting these signs died within the 96-hr test period.<\/jats:p>\n                  <jats:p>Furthermore, the survival times from the onset of these signs to death were sufficiently short to justify their definition as moribund states. Accordingly, we defined these signs as moribund endpoints and propose that fish exhibiting any of them should be euthanized during the test period. Notably, these findings align with previously reported results in Japanese medaka, indicating the potential cross-species applicability of these moribund endpoints.<\/jats:p>","DOI":"10.1093\/etojnl\/vgaf288","type":"journal-article","created":{"date-parts":[[2025,12,5]],"date-time":"2025-12-05T09:11:15Z","timestamp":1764925875000},"page":"420-426","source":"Crossref","is-referenced-by-count":1,"title":["Defining moribund states in zebrafish to refine fish acute toxicity tests"],"prefix":"10.1093","volume":"45","author":[{"given":"Aiyi","family":"Sui","sequence":"first","affiliation":[{"name":"Department of Molecular and Developmental Biology, Institute of Medicine, University of Tsukuba , Tsukuba,","place":["Japan"]},{"name":"Doctoral Program in Medical Science, Degree Programs in Comprehensive Human Sciences, Graduate School of Comprehensive Human Sciences, University of Tsukuba , Tsukuba,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Riping","family":"Huang","sequence":"additional","affiliation":[{"name":"Health and Environmental Risk Division, National Institute for Environmental Studies , Tsukuba,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takahiro","family":"Yamagishi","sequence":"additional","affiliation":[{"name":"Health and Environmental Risk Division, National Institute for Environmental Studies , Tsukuba,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Makoto","family":"Kobayashi","sequence":"additional","affiliation":[{"name":"Department of Molecular and Developmental Biology, Institute of Medicine, University of Tsukuba , Tsukuba,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2025,12,4]]},"reference":[{"key":"2026020503480718200_vgaf288-B1","doi-asserted-by":"publisher","first-page":"2076","DOI":"10.1002\/etc.4824","article-title":"Key opportunities to replace, reduce, and refine regulatory fish acute toxicity tests","volume":"39","author":"Burden","year":"2020","journal-title":"Environmental Toxicology and Chemistry"},{"key":"2026020503480718200_vgaf288-B2","doi-asserted-by":"publisher","first-page":"677","DOI":"10.1016\/j.chemosphere.2017.08.047","article-title":"Fish embryo toxicity test, threshold approach, and moribund as approaches to implement 3R principles to the acute fish toxicity 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