{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T10:14:41Z","timestamp":1785406481316,"version":"3.56.0"},"posted":{"date-parts":[[2026,2,6]]},"group-title":"Zoology","reference-count":56,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,2,6]],"date-time":"2026-02-06T00:00:00Z","timestamp":1770336000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"award":["NIH NIGMS R35GM150574"],"award-info":[{"award-number":["NIH NIGMS R35GM150574"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,2,6]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Non-lethal sampling methods are increasingly essential for amphibian research as global declines intensify and many species persist in small, vulnerable populations. Skin biopsies offer a promising alternative to whole-animal collection and other minimally invasive approaches; however, systematic evaluations of recovery and impacts on body condition remain limited. Here, we assess the effects of small (2-mm) dorsal skin biopsies in four frog species, including three highly endangered Harlequin frogs (\n                  <jats:italic>Atelopus bomolochos<\/jats:italic>\n                  ,\n                  <jats:italic>A. balios<\/jats:italic>\n                  ,\n                  <jats:italic>A. longirostris<\/jats:italic>\n                  ) and the Gualataco marsupial frog (\n                  <jats:italic>Gastrotheca riobambae<\/jats:italic>\n                  ). Under controlled laboratory conditions and in semi-natural enclosures, we monitored wound healing, survival, and body mass trajectories in biopsied and control individuals over a one-month period. Across all species, biopsy sites fully healed within approximately three weeks, following consistent stages of re-epithelialization and subsequent repigmentation. No biopsy-related mortality was observed, and body mass did not differ between biopsied and control individuals, indicating no detectable effects of skin biopsies on body condition during the wound-healing period. Occasional minor post-biopsy reactions resolved without intervention within the observation period. We additionally report anecdotal field recovery observations for three other species (\n                  <jats:italic>A. coynei<\/jats:italic>\n                  ,\n                  <jats:italic>A. laetissimus<\/jats:italic>\n                  , and\n                  <jats:italic>A.<\/jats:italic>\n                  sp.\n                  <jats:italic>aff. longirostris<\/jats:italic>\n                  ), indicating survival and visible wound closure following release. Together, these results indicate that small dorsal skin biopsies represent a safe, non-lethal sampling method for amphibians, including highly endangered taxa. By providing sufficient tissue for diverse downstream applications\u2014such as chemical analyses, genomics, transcriptomics, microbiome characterization, and disease detection\u2014this approach expands the range of questions that can be addressed while minimizing harm to threatened species.\n                <\/jats:p>","DOI":"10.64898\/2026.02.04.703782","type":"posted-content","created":{"date-parts":[[2026,2,7]],"date-time":"2026-02-07T02:55:19Z","timestamp":1770432919000},"source":"Crossref","is-referenced-by-count":0,"title":["Dorsal skin biopsies: A non-lethal sampling method for studying amphibians, including the highly endangered Harlequin frogs (Bufonidae:\n                  <i>Atelopus<\/i>\n                  )"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1455-6352","authenticated-orcid":false,"given":"Mar\u00eda Jos\u00e9 Navarrete","family":"M\u00e9ndez","sequence":"first","affiliation":[{"name":"Department of Integrative Biology and Museum of Vertebrate Zoology, University of California Berkeley, Berkeley, CA, USA 94720"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amanda B. Quezada","family":"Riera","sequence":"additional","affiliation":[{"name":"Centro Jambatu de Investigaci\u00f3n y Conservaci\u00f3n de Anfibios, Fundaci\u00f3n Jambatu, Giovanni Farina 566 y Baltra, San Rafael, Quito, Ecuador"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrea","family":"Ter\u00e1n-Valdez","sequence":"additional","affiliation":[{"name":"Centro Jambatu de Investigaci\u00f3n y Conservaci\u00f3n de Anfibios, Fundaci\u00f3n Jambatu, Giovanni Farina 566 y Baltra, San Rafael, Quito, Ecuador"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elena","family":"Naydenova","sequence":"additional","affiliation":[{"name":"Centro Jambatu de Investigaci\u00f3n y Conservaci\u00f3n de Anfibios, Fundaci\u00f3n Jambatu, Giovanni Farina 566 y Baltra, San Rafael, Quito, Ecuador"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luis A.","family":"Coloma","sequence":"additional","affiliation":[{"name":"Centro Jambatu de Investigaci\u00f3n y Conservaci\u00f3n de Anfibios, Fundaci\u00f3n Jambatu, Giovanni Farina 566 y Baltra, San Rafael, Quito, Ecuador"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5387-7250","authenticated-orcid":false,"given":"Rebecca D.","family":"Tarvin","sequence":"additional","affiliation":[{"name":"Department of Integrative Biology and Museum of Vertebrate Zoology, University of California Berkeley, Berkeley, CA, USA 94720"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"issue":"2","key":"2026021008153967000_2026.02.04.703782v1.1","doi-asserted-by":"crossref","first-page":"249","DOI":"10.1163\/15685381-bja10130","article-title":"Buccal Swabs for Amphibian Genomics","volume":"44","year":"2023","journal-title":"Amphibia-Reptilia: Publication of the Societas Europaea Herpetologica"},{"issue":"March","key":"2026021008153967000_2026.02.04.703782v1.2","first-page":"87","article-title":"Quantifying Tetrodotoxin Levels in the California Newt Using a Non-Destructive Sampling Method","volume":"80","year":"2014","journal-title":"Toxicon: Official Journal of the International Society on Toxinology"},{"key":"2026021008153967000_2026.02.04.703782v1.3","doi-asserted-by":"publisher","DOI":"10.1007\/978-3-642-02421-4_7"},{"issue":"3","key":"2026021008153967000_2026.02.04.703782v1.4","first-page":"244","article-title":"Uses and Doses of Local Anesthetics in Fish, Amphibians, and Reptiles","volume":"56","year":"2017","journal-title":"Journal of the American Association for Laboratory Animal Science: JAALAS"},{"key":"2026021008153967000_2026.02.04.703782v1.5","doi-asserted-by":"crossref","unstructured":"Coloma, Luis A. , and William E. Duellman . 2025a. Amphibians of Ecuador: Phyllomedusidae, Leptodactylidae, Ceratophryidae, Hemiphractidae, Eleutherodactylidae, Centrolenidae, Gymnophiona and Caudata, Volume IV. 1st Edition. CRC Press.","DOI":"10.1201\/9781003543879-1"},{"key":"2026021008153967000_2026.02.04.703782v1.6","doi-asserted-by":"crossref","unstructured":"Coloma, Luis A. , and William E. Duellman . 2025b. Amphibians of Ecuador: Pipidae, Telmatobiidae, Microhylidae, Dendrobatidae, Ranidae, Bufonidae, and Hylidae, Volume II. 1st Edition. CRC Press.","DOI":"10.1201\/9781003543886-1"},{"key":"2026021008153967000_2026.02.04.703782v1.7","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1016\/j.envres.2018.03.044","article-title":"\u201cElectron Spin Resonance (ESR) for the Study of Reactive Oxygen Species (ROS) on the Isolated Frog Skin (Pelophylax bergeri): A Non-Invasive Method for Environmental Monitoring.\u201d","volume":"165","year":"2018","journal-title":"Environmental Research"},{"key":"2026021008153967000_2026.02.04.703782v1.8","doi-asserted-by":"crossref","unstructured":"Duellman, William E. , and Linda Trueb . 1994. Biology of Amphibians. Johns Hopkins University Press.","DOI":"10.56021\/9780801847806"},{"key":"2026021008153967000_2026.02.04.703782v1.9","unstructured":"Empresa P\u00fablica Metropolitana de Agua Potable y Saneamiento de Quito (EPMAPS). 2026. PARAM H2O: Plataforma de monitoreo hidrometeorol\u00f3gico de Quito. Accessed January 29, 2026. https:\/\/paramh2o.aguaquito.gob.ec\/."},{"key":"2026021008153967000_2026.02.04.703782v1.10","first-page":"381","article-title":"Pharmacology and Chemistry of Chiriquitoxin a New Tetrodotoxin-like Substance from the Costa Rican Frog Atelopus chiriquiensis","volume":"19","year":"1976","journal-title":"Proceedings of the Western Pharmacology Society"},{"issue":"1713361","key":"2026021008153967000_2026.02.04.703782v1.11","first-page":"1713361","article-title":"The Contribution of Amphibian Macrophage Subsets to Scarless Regeneration of Skin Wounds","volume":"16","year":"2025","journal-title":"Frontiers in Immunology"},{"issue":"4","key":"2026021008153967000_2026.02.04.703782v1.12","doi-asserted-by":"crossref","first-page":"650","DOI":"10.1643\/CH-14-064","article-title":"Toe Clipping Does Not Affect the Survival of Leopard Frogs (Rana pipiens)","volume":"2014","year":"2014","journal-title":"Copeia"},{"key":"2026021008153967000_2026.02.04.703782v1.13","doi-asserted-by":"publisher","DOI":"10.1016\/j.diff.2014.02.002"},{"issue":"1","key":"2026021008153967000_2026.02.04.703782v1.14","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1163\/156853894X00560","article-title":"Inflammation due to Toe-Clipping in Natterjack Toads (Bufo calamita)","volume":"15","year":"1994","journal-title":"Amphibia-Reptilia: Publication of the Societas Europaea Herpetologica"},{"issue":"1","key":"2026021008153967000_2026.02.04.703782v1.15","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1670\/10-016.1","article-title":"Putting Toe Clipping into Perspective: A Viable Method for Marking Anurans","volume":"45","year":"2011","journal-title":"Journal of Herpetology"},{"issue":"8","key":"2026021008153967000_2026.02.04.703782v1.16","doi-asserted-by":"crossref","first-page":"1149","DOI":"10.1016\/S0041-0101(02)00115-0","article-title":"Tetrodotoxin Levels of the Rough-Skin Newt, Taricha granulosa, Increase in Long-Term Captivity","volume":"40","year":"2002","journal-title":"Toxicon: Official Journal of the International Society on Toxinology"},{"key":"2026021008153967000_2026.02.04.703782v1.17","doi-asserted-by":"publisher","DOI":"10.1023\/A:1021049125805"},{"key":"2026021008153967000_2026.02.04.703782v1.18","unstructured":"Hardman, Rebecca . 2020. \u201cEvaluation of Skin Health in Hellbenders, Cryptobranchus alleganiensis.\u201d University of Tennessee, Knoxville. https:\/\/trace.tennessee.edu\/utk_graddiss\/5861."},{"issue":"1","key":"2026021008153967000_2026.02.04.703782v1.19","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1655\/HERPMONOGRAPHS-D-14-00006","article-title":"Occurrence of Amphibians in Saline Habitats: A Review and Evolutionary Perspective","volume":"29","year":"2015","journal-title":"Herpetological Monograph"},{"key":"2026021008153967000_2026.02.04.703782v1.20","doi-asserted-by":"crossref","unstructured":"Hua, Jessica , Devin K. Jones , Brian M. Mattes , Rickey D. Cothran , Rick A. Relyea , and Jason T. Hoverman . 2015. \u201cEvolved Pesticide Tolerance in Amphibians: Predicting Mechanisms Based on Pesticide Novelty and Mode of Action.\u201d Environmental Pollution (Barking, Essex: 1987) 206 (November): 56\u201363.","DOI":"10.1016\/j.envpol.2015.06.030"},{"issue":"12","key":"2026021008153967000_2026.02.04.703782v1.21","doi-asserted-by":"crossref","first-page":"1892","DOI":"10.3390\/biomedicines9121892","article-title":"Skin Wound Healing of the Adult Newt, Cynops pyrrhogaster: A Unique Re-Epithelialization and Scarless Model","volume":"9","year":"2021","journal-title":"Biomedicines"},{"key":"2026021008153967000_2026.02.04.703782v1.22","unstructured":"IUCN Red List of Threatened Species. 2025. \u201cThe IUCN Red List of Threatened Species.\u201d Accessed April 18, 2025. https:\/\/www.iucnredlist.org\/en."},{"key":"2026021008153967000_2026.02.04.703782v1.23","doi-asserted-by":"publisher","DOI":"10.2305\/IUCN.UK.2018-1.RLTS.T54491A98640710.en"},{"key":"2026021008153967000_2026.02.04.703782v1.24","doi-asserted-by":"publisher","DOI":"10.2305\/IUCN.UK.2019-3.RLTS.T54492A98640844.en"},{"key":"2026021008153967000_2026.02.04.703782v1.25","doi-asserted-by":"publisher","DOI":"10.2305\/IUCN.UK.2022-1.RLTS.T54522A98641996.en"},{"key":"2026021008153967000_2026.02.04.703782v1.26","doi-asserted-by":"publisher","DOI":"10.2305\/IUCN.UK.2023-1.RLTS.T55357A98647877.en"},{"key":"2026021008153967000_2026.02.04.703782v1.27","doi-asserted-by":"publisher","DOI":"10.1126\/science.1138374"},{"key":"2026021008153967000_2026.02.04.703782v1.28","doi-asserted-by":"publisher","DOI":"10.1016\/S0195-5616(08)70010-X"},{"issue":"1","key":"2026021008153967000_2026.02.04.703782v1.29","doi-asserted-by":"crossref","first-page":"412","DOI":"10.1038\/s43247-023-01069-w","article-title":"Ongoing Harlequin Toad Declines Suggest the Amphibian Extinction Crisis Is Still an Emergency","volume":"4","year":"2023","journal-title":"Communications Earth & Environment"},{"issue":"1763","key":"2026021008153967000_2026.02.04.703782v1.30","first-page":"20170386","article-title":"Biological Collections for Understanding Biodiversity in the Anthropocene","volume":"374","year":"2018","journal-title":"Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences"},{"key":"2026021008153967000_2026.02.04.703782v1.31","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2123070119"},{"key":"2026021008153967000_2026.02.04.703782v1.32","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.3002318"},{"key":"2026021008153967000_2026.02.04.703782v1.33","doi-asserted-by":"publisher","DOI":"10.1016\/S0306-4565(97)00065-X"},{"issue":"3","key":"2026021008153967000_2026.02.04.703782v1.34","first-page":"266","article-title":"Effect of Toe-Clipping on the Survival of Several Lizard Species","volume":"27","year":"2017","journal-title":"Herpetological Journal"},{"issue":"20","key":"2026021008153967000_2026.02.04.703782v1.35","doi-asserted-by":"crossref","first-page":"3162","DOI":"10.3390\/ani13203162","article-title":"Current State of Conservation Physiology for Amphibians: Major Research Topics and Physiological Parameters","volume":"13","year":"2023","journal-title":"Animals: An Open Access Journal from MDPI"},{"issue":"3","key":"2026021008153967000_2026.02.04.703782v1.36","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1163\/156853801317050070","article-title":"Identifying Effects of Toe Clipping on Anuran Return Rates: The Importance of Statistical Power","volume":"22","year":"2001","journal-title":"Amphibia-Reptilia: Publication of the Societas Europaea Herpetologica"},{"issue":"12","key":"2026021008153967000_2026.02.04.703782v1.37","first-page":"001927","article-title":"A Novel Tamanavirus (Flaviviridae) of the European Common Frog (Rana temporaria) from the UK","volume":"104","year":"2023","journal-title":"The Journal of General Virology"},{"issue":"5","key":"2026021008153967000_2026.02.04.703782v1.38","doi-asserted-by":"crossref","first-page":"752","DOI":"10.1111\/1749-4877.12584","article-title":"Global Gaps in Age Data Based on Skeletochronology for Amphibians","volume":"17","year":"2022","journal-title":"Integrative Zoology"},{"issue":"4","key":"2026021008153967000_2026.02.04.703782v1.39","doi-asserted-by":"crossref","first-page":"547","DOI":"10.1670\/11-037.1","article-title":"Toe Clipping of Amphibians and Reptiles: Science, Ethics, and the Law","volume":"45","year":"2011","journal-title":"Journal of Herpetology"},{"issue":"3","key":"2026021008153967000_2026.02.04.703782v1.40","first-page":"305","article-title":"Toe-Clipping as an Acceptable Method of Identifying Individual Anurans in Mark Recapture Studies","volume":"38","year":"2007","journal-title":"Herpetological Review"},{"issue":"3","key":"2026021008153967000_2026.02.04.703782v1.41","first-page":"204","article-title":"\u201cInflammation in Digits of Unmarked and Toe-Tipped Wild Hylids.\u201d Wildlife Research (East Melbourne, Melbourne","volume":"38","year":"2011","journal-title":"Vic"},{"key":"2026021008153967000_2026.02.04.703782v1.42","unstructured":"Posit Team. 2025. RStudio: Integrated Development Environment for R. Version 2025.5.0.496. MacOS."},{"issue":"1","key":"2026021008153967000_2026.02.04.703782v1.43","doi-asserted-by":"crossref","first-page":"bpae030","DOI":"10.1093\/biomethods\/bpae030","article-title":"Comparing Skin Swabs, Buccal Swabs, and Toe Clips for Amphibian Genetic Sampling, a Case Study with a Small Anuran (Acris blanchardi)","volume":"9","year":"2024","journal-title":"Biology Methods and Protocols"},{"issue":"2","key":"2026021008153967000_2026.02.04.703782v1.44","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1163\/15685381-17000206","article-title":"Testing Skin Swabbing for DNA Sampling in Dendrobatid Frogs","volume":"39","year":"2018","journal-title":"Amphibia-Reptilia: Publication of the Societas Europaea Herpetologica"},{"key":"2026021008153967000_2026.02.04.703782v1.45","unstructured":"Ron, S. R. , Merino-Viteri, A. Ortiz, D. A. 2024. Anfibios del Ecuador. Version 2024.0. Museo de Zoolog\u00eda, Pontificia Universidad Cat\u00f3lica del Ecuador. < https:\/\/bioweb.bio\/faunaweb\/amphibiaweb>, fecha de acceso 14 de diciembre, 2025. AmphibiaWeb. 2025. <https:\/\/amphibiaweb.org> University of California, Berkeley, CA, USA. Accessed 14 Dec 2025."},{"key":"2026021008153967000_2026.02.04.703782v1.46","doi-asserted-by":"publisher","DOI":"10.1002\/jez.1402160311"},{"issue":"5702","key":"2026021008153967000_2026.02.04.703782v1.47","doi-asserted-by":"crossref","first-page":"1783","DOI":"10.1126\/science.1103538","article-title":"\u201cStatus and Trends of Amphibian Declines and Extinctions Worldwide.\u201d Science (New York","volume":"306","year":"2004","journal-title":"N.Y"},{"key":"2026021008153967000_2026.02.04.703782v1.48","doi-asserted-by":"publisher","DOI":"10.1641\/0006-3568(2004)054[0066:TVOMCF]2.0.CO;2"},{"issue":"4597","key":"2026021008153967000_2026.02.04.703782v1.49","doi-asserted-by":"crossref","first-page":"609","DOI":"10.1126\/science.6573024","article-title":"\u201cInhibition of Gastric Acid Secretion in the Gastric Brooding Frog, Rheobatrachus silus.\u201d Science (New York","volume":"220","year":"1983","journal-title":"N.Y"},{"key":"2026021008153967000_2026.02.04.703782v1.50","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.50678"},{"issue":"3","key":"2026021008153967000_2026.02.04.703782v1.51","doi-asserted-by":"crossref","first-page":"467","DOI":"10.1670\/07-265.1","article-title":"Modeling the Effect of Toe Clipping on Treefrog Survival: Beyond the Return Rate","volume":"42","year":"2008","journal-title":"Journal of Herpetology"},{"key":"2026021008153967000_2026.02.04.703782v1.52","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1716257115"},{"issue":"4","key":"2026021008153967000_2026.02.04.703782v1.53","doi-asserted-by":"crossref","first-page":"638","DOI":"10.1643\/h2022005","article-title":"State of the Amphibia 2020: A Review of Five Years of Amphibian Research and Existing Resources","volume":"110","year":"2022","journal-title":"Ichthyology & Herpetology"},{"issue":"2","key":"2026021008153967000_2026.02.04.703782v1.54","first-page":"79","article-title":"Comparative Skin Histology of Frogs Reveals High-Elevation Adaptation of the Tibetan Nanorana parkeri","volume":"10","year":"2019","journal-title":"Asian Herpetological Research"},{"issue":"22","key":"2026021008153967000_2026.02.04.703782v1.55","doi-asserted-by":"crossref","first-page":"3187","DOI":"10.1016\/S0040-4039(00)94728-2","article-title":"The Structure of Chiriquitoxin from the Costa Rican Frog","volume":"31","year":"1990","journal-title":"Tetrahedron Letters"},{"issue":"e03582","key":"2026021008153967000_2026.02.04.703782v1.56","first-page":"e03582","article-title":"The Impact of Toe-Clipping on Animal Welfare in Amphibians: A Systematic Review","volume":"59","year":"2025","journal-title":"Global Ecology and Conservation"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.64898\/2026.02.04.703782","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,2,10]],"date-time":"2026-02-10T16:15:54Z","timestamp":1770740154000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.64898\/2026.02.04.703782"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,2,6]]},"references-count":56,"URL":"https:\/\/doi.org\/10.64898\/2026.02.04.703782","relation":{},"subject":[],"published":{"date-parts":[[2026,2,6]]},"subtype":"preprint"}}