{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,9,20]],"date-time":"2025-09-20T04:33:50Z","timestamp":1758342830542,"version":"3.44.0"},"reference-count":54,"publisher":"Public Library of Science (PLoS)","issue":"9","license":[{"start":{"date-parts":[[2025,9,19]],"date-time":"2025-09-19T00:00:00Z","timestamp":1758240000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"NIH-NIGMS Minnesota IRACDA Program","award":["2K12GM119955"],"award-info":[{"award-number":["2K12GM119955"]}]},{"name":"NSF STEM-APWD DEB","award":["DEB2316784"],"award-info":[{"award-number":["DEB2316784"]}]},{"name":"NIH","award":["R01-HL121212"],"award-info":[{"award-number":["R01-HL121212"]}]},{"name":"NIH","award":["R35-HL171421"],"award-info":[{"award-number":["R35-HL171421"]}]},{"name":"NIH","award":["R35HL150784"],"award-info":[{"award-number":["R35HL150784"]}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:p>Interactions between predators and prey are often characterized by strong selection pressures that shape extreme physiological adaptations. Venom resistance in large-bodied South American opossums (Clade Didelphini) is a striking example, as these marsupials prey on venomous snakes and exhibit remarkable resistance to their venom. While resistance is well documented in Didelphini, relatively little is known about venom resistance in the smaller, more diverse members of Didelphidae, which inhabit the same regions and encounter the same predators. Moreover, resistance of opossum von Willebrand factor (VWF) to the venom C-type lectin-like proteins has not been previously studied under simulated vascular flow states. Here, we use microfluidic devices to investigate venom resistance in the small-bodied opossum, <jats:italic>Monodelphis domestica,<\/jats:italic> examining platelet adhesion and fibrin deposition in response to purified venom components. Additionally, we conduct platelet aggregometry and assays of serum protease inhibitors in the presence of venom from sympatric and allopatric vipers to examine patterns of species-specificity and adaptation. Our results show that <jats:italic>M. domestica<\/jats:italic> resists venom-induced disruptions to platelet function in the presence of platelet-disrupting venom components botrocetin and convulxin, while aspercetin disrupts platelet and fibrin function similarly in opossum and human samples. Whole blood aggregometry and serum protease inhibition showed patterns consistent with species-specific adaptation of mammals to their local snake venom. Unexpectedly, we find that <jats:italic>M. domestica<\/jats:italic> VWF requires increased shear force to elongate, a previously unknown aspect of opossum blood physiology that may contribute to venom resistance and may have relevance to human coagulopathies. Our findings demonstrate resistance under natural shear stress, and document venom resistance beyond large-bodied Didelphini, suggesting it is a widespread trait in South American marsupials.<\/jats:p>","DOI":"10.1371\/journal.pone.0332686","type":"journal-article","created":{"date-parts":[[2025,9,19]],"date-time":"2025-09-19T17:25:38Z","timestamp":1758302738000},"page":"e0332686","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["Comparative microfluidic and enzymatic analyses reveal multifaceted snake venom resistance and novel VWF behavior in the opossum Monodelphis domestica"],"prefix":"10.1371","volume":"20","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3477-3012","authenticated-orcid":true,"given":"Matthew L.","family":"Holding","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dante","family":"Disharoon","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2237-659X","authenticated-orcid":true,"given":"Laura M.","family":"Haynes","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bipin","family":"Chakravarthy Paruchuri","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"M. Hao Hao","family":"Pontius","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Krista","family":"Golden","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2874-4904","authenticated-orcid":true,"given":"Jordan A.","family":"Shavit","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Karl C.","family":"Desch","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David","family":"Ginsburg","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anirban","family":"Sen Gupta","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yolanda","family":"Cruz","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7096-3254","authenticated-orcid":true,"given":"Danielle H.","family":"Drabeck","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2025,9,19]]},"reference":[{"issue":"1","key":"pone.0332686.ref001","doi-asserted-by":"crossref","first-page":"219","DOI":"10.1146\/annurev.es.25.110194.001251","article-title":"The evolutionary interaction among species: selection, escalation, and coevolution","volume":"25","author":"GJ Vermeij","year":"1994","journal-title":"Annu Rev Ecol Syst"},{"issue":"6157","key":"pone.0332686.ref002","doi-asserted-by":"crossref","first-page":"441","DOI":"10.1126\/science.1236451","article-title":"Voltage-gated sodium channel in grasshopper mice defends against bark scorpion toxin","volume":"342","author":"AH Rowe","year":"2013","journal-title":"Science"},{"issue":"11","key":"pone.0332686.ref003","doi-asserted-by":"crossref","first-page":"1469","DOI":"10.1517\/14712598.2011.621940","article-title":"Venoms as a platform for human drugs: translating toxins into therapeutics","volume":"11","author":"GF King","year":"2011","journal-title":"Expert Opin Biol Ther"},{"issue":"5","key":"pone.0332686.ref004","doi-asserted-by":"crossref","first-page":"1032","DOI":"10.1093\/icb\/icw082","article-title":"Venom resistance as a model for understanding the molecular basis of complex coevolutionary adaptations","volume":"56","author":"ML Holding","year":"2016","journal-title":"Integr Comp Biol"},{"key":"pone.0332686.ref005","first-page":"557","volume-title":"The ecological and evolutionary context of mammalian resistance to rattlesnake venoms. 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IUCN Red List of Threatened Species: Crotalus adamanteus. IUCN Red List of Threatened Species. 2007 [cited 26 Jun 2025]. Available from: https:\/\/www.iucnredlist.org\/en"},{"key":"pone.0332686.ref040","unstructured":"Santiago Carreira (Secci\u00f3n Zoolog\u00eda Vertebrados F de C& MN de HN\/ M-U, Lillo-CONICET) RM (Fundaci\u00f3n M, Paraguay) PC (Instituto de IB del, Plata-CONICET) FK (Museo de L, C\u00f3rdoba) NP (Universidad N de, Assessment) NS (SRLI R. IUCN Red List of Threatened Species: Bothrops jararaca. IUCN Red List of Threatened Species. 2014 [cited 26 Jun 2025]. Available from: https:\/\/www.iucnredlist.org\/en"},{"key":"pone.0332686.ref041","unstructured":"University N de la S (Texas T, Rivadavia) DF (Museo A de CNB. IUCN Red List of Threatened Species: Monodelphis domestica. IUCN Red List of Threatened Species. 2016 [cited 26 June 2025]. 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