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This makes the identification of physiological stress indicators crucial to assessing hydric risk in amphibian populations. This study examines iris darkening as a response to dehydration in the neotropical tree frog (\n                    <jats:italic>Boana punctata<\/jats:italic>\n                    ) under controlled conditions. Twenty\u2010five adult frogs were collected in the province of Chaco, Argentina, and subjected to sequential dehydration treatments (10, 20, and 30\u2009min) using two methods (constant airflow of 0.2\u2009m\/s or desiccant rice), followed by 2\u2010h rehydration. Linear mixed\u2011effects models revealed a strong negative relationship between hydration level and iris darkening, increasing about 1.5% in dark area for each 1% loss in hydration level, consistent across dehydration methods and with similar sensitivity across body sizes and individuals, although larger frogs showed lower baseline darkening. Iris darkening showed partial reversal post\u2010rehydration. This validates iris darkening as a robust, non\u2010invasive biomarker for hydric stress in wild amphibian populations facing climate\u2010driven aridity.\n                  <\/jats:p>","DOI":"10.1002\/jez.70118","type":"journal-article","created":{"date-parts":[[2026,7,27]],"date-time":"2026-07-27T08:02:00Z","timestamp":1785139320000},"page":"887-893","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Eye Darkening Under Dehydration Stress in the Neotropical Frog\n                    <i>Boana punctata<\/i>\n                    (Schneider, 1799)"],"prefix":"10.1002","volume":"345","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3947-8442","authenticated-orcid":false,"given":"Ricardo","family":"Reques","sequence":"first","affiliation":[{"name":"Departamento de Ecolog\u00eda y Evoluci\u00f3n Estaci\u00f3n Biol\u00f3gica de Do\u00f1ana\u2010CSIC Sevilla Spain"},{"name":"Departamento de Zoolog\u00eda, Facultad de Ciencias Universidad de C\u00f3rdoba C\u00f3rdoba Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1739-976X","authenticated-orcid":false,"given":"Eduardo F.","family":"Schaefer","sequence":"additional","affiliation":[{"name":"Instituto de Investigaciones Geohist\u00f3ricas (IIGHI \u2013 UNNE) Resistencia Chaco Argentina"},{"name":"Consejo Nacional de Investigaciones Cient\u00edficas y T\u00e9cnicas (CONICET) Buenos Aires Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3981-045X","authenticated-orcid":false,"given":"Eduardo A.","family":"Sanabria","sequence":"additional","affiliation":[{"name":"Consejo Nacional de Investigaciones Cient\u00edficas y T\u00e9cnicas (CONICET) Buenos Aires Argentina"},{"name":"Instituto de Ciencias B\u00e1sicas, Facultad de Filosof\u00eda Humanidades y Artes Universidad Nacional de San Juan San Juan Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-4139-7002","authenticated-orcid":false,"given":"Leonardo D.","family":"Aguiar","sequence":"additional","affiliation":[{"name":"Consejo Nacional de Investigaciones Cient\u00edficas y T\u00e9cnicas (CONICET) Buenos Aires Argentina"},{"name":"Instituto de Bot\u00e1nica del Nordeste Universidad Nacional del Nordeste Corrientes Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0065-750X","authenticated-orcid":false,"given":"Lorena B.","family":"Quiroga","sequence":"additional","affiliation":[{"name":"Consejo Nacional de Investigaciones Cient\u00edficas y T\u00e9cnicas (CONICET) Buenos Aires Argentina"},{"name":"Instituto de Ciencias B\u00e1sicas, Facultad de Filosof\u00eda Humanidades y Artes Universidad Nacional de San Juan San Juan Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4183-184X","authenticated-orcid":false,"given":"Miguel","family":"Tejedo","sequence":"additional","affiliation":[{"name":"Departamento de Ecolog\u00eda y Evoluci\u00f3n Estaci\u00f3n Biol\u00f3gica de Do\u00f1ana\u2010CSIC Sevilla Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,7,27]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1007\/s003590050023"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1086\/690189"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/0016-6480(69)90052-5"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1002\/jez.2250"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.3390\/d10040118"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1670\/13-206"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.zool.2013.09.005"},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.1098\/rspb.2020.2273"},{"key":"e_1_2_10_10_1","doi-asserted-by":"publisher","DOI":"10.1210\/endo-84-1-69"},{"key":"e_1_2_10_11_1","doi-asserted-by":"publisher","DOI":"10.1152\/ajpregu.00160.2018"},{"key":"e_1_2_10_12_1","doi-asserted-by":"crossref","unstructured":"IUCN SSC Amphibian Specialist Group.2023. \u201cBoana punctata.\u201d In The IUCN Red List of Threatened Species 2023: e.T55620A61401241.https:\/\/doi.org\/10.2305\/IUCN.UK.2023-1.RLTS.T55620A61401241.en.","DOI":"10.2305\/IUCN.UK.2023-1.RLTS.T55620A61401241.en"},{"key":"e_1_2_10_13_1","doi-asserted-by":"publisher","DOI":"10.1017\/S0952836904004923"},{"key":"e_1_2_10_14_1","doi-asserted-by":"crossref","unstructured":"Lenth R. 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