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Our knowledge of parasite cyst stage conversion and interaction with the intestinal tract is limited. Here, we investigate infection dynamics of <jats:italic>Toxoplasma gondii<\/jats:italic> cyst-stage in murine jejunum and human intestinal microphysiological systems. We focus on parasite ingress, replication, and conversion of the cyst stage to the rapidly replicating dissemination stage. <jats:italic>In vivo<\/jats:italic> bioluminescent imaging of mice fed cysts revealed spots of infection throughout the jejunum and ileum, which were selected for further analyses. Immunostaining showed parasite migration and replication predominantly in the stroma, with minimal replication in enterocytes. We recapitulated bradyzoite infection in human intestinal microphysiological systems and showed stage conversation and migration through collagen. This integrated approach elucidates complex host-parasite interactions, highlighting the value of microphysiological systems in advancing understanding and identifying potential therapeutics.<\/jats:p>","DOI":"10.1371\/journal.pntd.0012855","type":"journal-article","created":{"date-parts":[[2025,2,4]],"date-time":"2025-02-04T18:39:43Z","timestamp":1738694383000},"page":"e0012855","update-policy":"https:\/\/doi.org\/10.1371\/journal.pntd.corrections_policy","source":"Crossref","is-referenced-by-count":3,"title":["Modeling Toxoplasma gondii-gut early interactions using a human microphysiological system"],"prefix":"10.1371","volume":"19","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1485-3499","authenticated-orcid":true,"given":"Carlos J.","family":"Ram\u00edrez-Flores","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicole D.","family":"Hryckowian","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrew N.","family":"Gale","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kehinde Adebayo","family":"Babatunde","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marcos","family":"Lares","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David J.","family":"Beebe","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sheena C.","family":"Kerr","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9362-8088","authenticated-orcid":true,"given":"Laura J.","family":"Knoll","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2025,2,4]]},"reference":[{"issue":"2","key":"pntd.0012855.ref001","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1007\/s12639-024-01664-4","article-title":"Toxoplasmosis in animals and humans: a neglected zoonotic disease in Bangladesh","volume":"48","author":"MF Hasan","year":"2024","journal-title":"J Parasit Dis"},{"issue":"1","key":"pntd.0012855.ref002","article-title":"Toxoplasmosis of animals and humans","volume":"3","author":"JP. 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A Toxoplasma gondii model to dissect bradyzoite recredescence in vitro (Master of Science thesis dissertation) [Internet]. The University of Georgia. https:\/\/getd.libs.uga.edu\/pdfs\/salas-ortega_omar_a_201908_ms.pdf; 2011. 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