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P\u2010deficient fish have been reported to exhibit morphometric alterations, notably in the ribs and neural\/hemal spines; however, whether these deformities represent true pathological manifestations remains uncertain. This study re\u2010examined skeletal tissues of juvenile rainbow trout\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>Oncorhynchus mykiss<\/jats:italic>\n                    <\/jats:styled-content>\n                    using alternative protocols. Fish were fed either a P\u2010deficient or a control diet for 1.5\u2009months. Following formalin fixation, muscular tissues were enzymatically digested, and the skeletons were stained with alizarin red, alcian blue and aniline blue for detailed examination. P\u2010deficient fish exhibited only minor rib deformities but showed extensive non\u2010calcified regions across the skeleton, teeth, fins and scales. These findings were further confirmed by scanning electron microscopy coupled with energy\u2010dispersive X\u2010ray spectroscopy (SEM\u2010EDS). In contrast, heating the skeletons (70\u00b0C or 100\u00b0C) markedly exacerbated skeletal deformities in P\u2010deficient fish but not in control fish. These results indicate that bending deformities previously reported as clinical signs of P deficiency are not evident in intact fish but become conspicuous only after heating during analytical procedures. The expansion of non\u2010calcified skeletal regions may serve as a sensitive diagnostic indicator of P deficiency in farmed fish.\n                  <\/jats:p>","DOI":"10.1111\/jfd.70172","type":"journal-article","created":{"date-parts":[[2026,3,27]],"date-time":"2026-03-27T05:26:34Z","timestamp":1774589194000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Macroscopic Examination of Skeletal Tissues in Phosphorus\u2010Deficient Rainbow Trout\n                    <i>Oncorhynchus mykiss<\/i>"],"prefix":"10.1111","volume":"49","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2292-2087","authenticated-orcid":false,"given":"Shozo H.","family":"Sugiura","sequence":"first","affiliation":[{"name":"Graduate School of Environmental Sciences The University of Shiga Prefecture  Hikone\u2010city Shiga Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,3,26]]},"reference":[{"key":"e_1_2_11_2_1","doi-asserted-by":"publisher","DOI":"10.3390\/polym13132151"},{"key":"e_1_2_11_3_1","doi-asserted-by":"publisher","DOI":"10.29309\/TPMJ\/2022.29.08.7023"},{"issue":"2","key":"e_1_2_11_4_1","first-page":"76","article-title":"Modified Double Skeletal Staining Protocols With Alizarinred and Alcian Blue in Laboratory Animals","volume":"13","author":"Aliesfehani T.","year":"2015","journal-title":"Annals of Military & Health Sciences Research"},{"key":"e_1_2_11_5_1","doi-asserted-by":"publisher","DOI":"10.1002\/jbmr.4486"},{"key":"e_1_2_11_6_1","doi-asserted-by":"publisher","DOI":"10.3390\/children12020144"},{"key":"e_1_2_11_7_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2095.1997.00069.x"},{"key":"e_1_2_11_8_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2095.1998.00095.x"},{"key":"e_1_2_11_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bpj.2011.04.033"},{"key":"e_1_2_11_10_1","doi-asserted-by":"publisher","DOI":"10.1002\/9780470697610.ch13"},{"key":"e_1_2_11_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ekir.2020.03.025"},{"key":"e_1_2_11_12_1","doi-asserted-by":"publisher","DOI":"10.1038\/jid.1968.67"},{"key":"e_1_2_11_13_1","doi-asserted-by":"publisher","DOI":"10.1093\/jbmrpl\/ziae081"},{"key":"e_1_2_11_14_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms21155429"},{"key":"e_1_2_11_15_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1749-7345.1987.tb00431.x"},{"key":"e_1_2_11_16_1","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-60327-483-8_5"},{"key":"e_1_2_11_17_1","doi-asserted-by":"publisher","DOI":"10.1111\/are.12549"},{"key":"e_1_2_11_18_1","doi-asserted-by":"publisher","DOI":"10.1177\/003591571600902050"},{"key":"e_1_2_11_19_1","doi-asserted-by":"publisher","DOI":"10.3109\/10520297709116780"},{"key":"e_1_2_11_20_1","doi-asserted-by":"publisher","DOI":"10.1177\/1925362118797737"},{"key":"e_1_2_11_21_1","doi-asserted-by":"publisher","DOI":"10.1002\/9780470697610.ch9"},{"key":"e_1_2_11_22_1","doi-asserted-by":"publisher","DOI":"10.1186\/s42825-024-00152-y"},{"issue":"3","key":"e_1_2_11_23_1","first-page":"143","article-title":"Kieferrachitis.\u201d Dtsch.\u2009Vjschr.\u2009Zahnheilk. 18:180\u2013185; Cited in: Seki, S., Tsuchida, S., Komoribayashi, N., Sekiyama, S., Nitanai, A. 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