{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T02:57:09Z","timestamp":1787021829476,"version":"build-2736575974"},"reference-count":98,"publisher":"Public Library of Science (PLoS)","issue":"7","license":[{"start":{"date-parts":[[2026,7,27]],"date-time":"2026-07-27T00:00:00Z","timestamp":1785110400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["www.plosntds.org"],"crossmark-restriction":false},"short-container-title":["PLoS Negl Trop Dis"],"abstract":"<jats:p>\n                    The study of snake venom toxicity and evaluation of the neutralisation ability of antivenoms and novel therapeutic agents for envenoming has largely relied on animal tests, especially the analysis of the lethal activity of venoms and its neutralisation. In addition, other animal-based assays are used to assess pathology-specific effects of venoms. Despite their demonstrated value, these assays have several limitations. They involve acute stress and pain in animals, and their validity,\n                    <jats:italic>vis-\u00e0-vis<\/jats:italic>\n                    the characteristics of human snakebite envenoming, is limited owing to the nature of the assays. Therefore, urgent innovations are required in this field. Ensuring the implementation of the 3Rs (Replacement, Reduction, and Refinement) is essential when animal studies are required. These include: (a) refinement of the tests (e.g., use of analgesia, reduction of assay duration, and refining of humane endpoints); (b) reduction in the number of animals used by more thorough\n                    <jats:italic>in vitro<\/jats:italic>\n                    assessment of antivenoms prior to\n                    <jats:italic>in vivo<\/jats:italic>\n                    assays, and improved statistical analyses and group sizing; and, most importantly, and (c) replacement of\n                    <jats:italic>in vivo<\/jats:italic>\n                    rodent tests with validated\n                    <jats:italic>in vitro<\/jats:italic>\n                    approaches, such as immunoassays, enzymatic and cell-based assays, the use of invertebrate models, and the implementation of New Approach Methodologies (NAMs), i.e., novel\n                    <jats:italic>in vitro<\/jats:italic>\n                    or\n                    <jats:italic>in silico<\/jats:italic>\n                    techniques that model complex pathophysiological processes of envenoming. A roadmap involving researchers, manufacturers, quality control groups, regulators, and funding agencies is proposed, with the aim of driving significant changes in this field.\n                  <\/jats:p>","DOI":"10.1371\/journal.pntd.0014537","type":"journal-article","created":{"date-parts":[[2026,7,27]],"date-time":"2026-07-27T17:30:32Z","timestamp":1785173432000},"page":"e0014537","update-policy":"https:\/\/doi.org\/10.1371\/journal.pntd.corrections_policy","source":"Crossref","is-referenced-by-count":1,"title":["Animal models in venom and antivenom research: The need to align academic discovery with manufacturing and regulatory expectations"],"prefix":"10.1371","volume":"20","author":[{"given":"Stuart","family":"Ainsworth","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicholas R.","family":"Casewell","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marco Aur\u00e9lio","family":"Sartim","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amy E.","family":"Marriott","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8385-3081","authenticated-orcid":true,"given":"Jos\u00e9 Mar\u00eda","family":"Guti\u00e9rrez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2026,7,27]]},"reference":[{"key":"pntd.0014537.ref001","doi-asserted-by":"crossref","first-page":"17063","DOI":"10.1038\/nrdp.2017.63","article-title":"Snakebite envenoming","volume":"3","author":"JM Guti\u00e9rrez","year":"2017","journal-title":"Nat Rev Dis Primers"},{"key":"pntd.0014537.ref002","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1016\/j.toxicon.2018.05.008","article-title":"The history of antivenoms development: beyond Calmette and Vital Brazil","volume":"150","author":"CC Squaiella-Baptist\u00e3o","year":"2018","journal-title":"Toxicon"},{"issue":"1","key":"pntd.0014537.ref003","doi-asserted-by":"crossref","first-page":"465","DOI":"10.1146\/annurev-pharmtox-022024-033544","article-title":"Progress and challenges in the field of snakebite envenoming therapeutics","volume":"65","author":"JM Guti\u00e9rrez","year":"2025","journal-title":"Annu Rev Pharmacol Toxicol"},{"issue":"6","key":"pntd.0014537.ref004","doi-asserted-by":"crossref","first-page":"799","DOI":"10.1080\/17460441.2025.2495943","article-title":"Practical progress towards the development of recombinant antivenoms for snakebite envenoming","volume":"20","author":"SK Menzies","year":"2025","journal-title":"Expert Opin Drug Discov"},{"key":"pntd.0014537.ref005","unstructured":"World Health Organization (WHO). 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