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In veterinary medicine, this apicomplexan parasite is considered one of the main infectious agents responsible for reproductive failure in small ruminants worldwide. The aim of this study was to phenotypically characterize 10 Spanish <jats:italic>T. gondii<\/jats:italic> isolates recently obtained from sheep in a normalized mouse model and in an ovine trophoblast cell line (AH-1) as infection target cells. The panel of isolates met selection criteria regarding such parameters as genetic diversity [types II (ToxoDB #1 and #3) and III (#2)], geographical location, and sample of origin (aborted foetal brain tissues or adult sheep myocardium). Evaluations of in vivo mortality, morbidity, parasite burden and histopathology were performed. Important variations between isolates were observed, although all isolates were classified as \u201cnonvirulent\u201d (&lt;\u200930% cumulative mortality). The isolates TgShSp16 (#3) and TgShSp24 (#2) presented higher degrees of virulence. Significant differences were found in terms of in vitro invasion rates and tachyzoite yield at 72\u00a0h post-inoculation (hpi) between TgShSp1 and TgShSp24 isolates, which exhibited the lowest and highest rates, respectively. The study of the <jats:italic>CS3<\/jats:italic>, <jats:italic>ROP18<\/jats:italic> and <jats:italic>ROP5<\/jats:italic> loci allelic profiles revealed only type III alleles in ToxoDB #2 isolates and type II alleles in the #1 and #3 isolates included. We concluded that there are relevant intra- and inter-genotype virulence differences in Spanish <jats:italic>T. gondii<\/jats:italic> isolates, which could not be inferred by genetic characterization using currently described molecular markers.<\/jats:p>","DOI":"10.1186\/s13567-021-00953-7","type":"journal-article","created":{"date-parts":[[2021,6,10]],"date-time":"2021-06-10T20:03:03Z","timestamp":1623355383000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":28,"title":["In vivo and in vitro models show unexpected degrees of virulence among Toxoplasma gondii type II and III isolates from sheep"],"prefix":"10.1186","volume":"52","author":[{"given":"Mercedes","family":"Fern\u00e1ndez-Escobar","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2323-0135","authenticated-orcid":false,"given":"Rafael","family":"Calero-Bernal","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Javier","family":"Regidor-Cerrillo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Raquel","family":"Vallejo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julio","family":"Benavides","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Esther","family":"Collantes-Fern\u00e1ndez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4986-6783","authenticated-orcid":false,"given":"Luis Miguel","family":"Ortega-Mora","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2021,6,10]]},"reference":[{"key":"953_CR1","volume-title":"Toxoplasmosis of animals and humans","author":"JP Dubey","year":"2010","unstructured":"Dubey JP (2010) Toxoplasmosis of animals and humans, 2nd edn. 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