{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,6]],"date-time":"2026-05-06T00:12:49Z","timestamp":1778026369123,"version":"3.51.4"},"reference-count":50,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2026,3,21]],"date-time":"2026-03-21T00:00:00Z","timestamp":1774051200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0"},{"start":{"date-parts":[[2026,5,6]],"date-time":"2026-05-06T00:00:00Z","timestamp":1778025600000},"content-version":"vor","delay-in-days":46,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0"}],"funder":[{"DOI":"10.13039\/501100004837","name":"Ministerio de Ciencia e Innovaci\u00f3n","doi-asserted-by":"publisher","award":["DIN2020-011454\/AEI\/10.13039\/501100011033"],"award-info":[{"award-number":["DIN2020-011454\/AEI\/10.13039\/501100011033"]}],"id":[{"id":"10.13039\/501100004837","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100012818","name":"Comunidad de Madrid","doi-asserted-by":"publisher","award":["Ref: S-2019\/L2-386"],"award-info":[{"award-number":["Ref: S-2019\/L2-386"]}],"id":[{"id":"10.13039\/100012818","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"DOI":"10.1038\/s41598-026-43786-0","type":"journal-article","created":{"date-parts":[[2026,3,21]],"date-time":"2026-03-21T06:23:45Z","timestamp":1774074225000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Development and validation of a gra1\u2013bag1 RT-qPCR assay as an alternative to the mouse bioassay for assessing Toxoplasma gondii viability"],"prefix":"10.1038","volume":"16","author":[{"given":"Andrea","family":"Largo-de la Torre","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Natalia","family":"Velasco-Jim\u00e9nez","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luis Miguel","family":"Ortega-Mora","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Javier","family":"Regidor-Cerrillo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2026,3,21]]},"reference":[{"key":"43786_CR1","volume-title":"Toxoplasmosis of animals and humans","author":"JP Dubey","year":"2022","unstructured":"Dubey, J. P. Toxoplasmosis of animals and humans 3rd ed. (CRC Press, 2022).","edition":"3rd ed"},{"key":"43786_CR2","doi-asserted-by":"publisher","first-page":"14","DOI":"10.1080\/20477724.2022.2083977","volume":"117","author":"S Dian","year":"2023","unstructured":"Dian, S., Ganiem, A. R. & Ekawardhani, S. Cerebral toxoplasmosis in HIV-infected patients: A review. Pathog. Glob. Health 117, 14\u201323. https:\/\/doi.org\/10.1080\/20477724.2022.2083977 (2023).","journal-title":"Pathog. Glob. Health"},{"key":"43786_CR3","doi-asserted-by":"publisher","first-page":"1079","DOI":"10.1007\/s10096-016-2656-2","volume":"35","author":"CP Rico-Torres","year":"2016","unstructured":"Rico-Torres, C. P., Vargas-Villavicencio, J. A. & Correa, D. Is Toxoplasma gondii type related to clinical outcome in human congenital infection? Systematic and critical review. Eur. J. Clin. Microbiol. Infect. Dis. 35, 1079\u20131088. https:\/\/doi.org\/10.1007\/s10096-016-2656-2 (2016).","journal-title":"Eur. J. Clin. Microbiol. Infect. Dis."},{"key":"43786_CR4","doi-asserted-by":"publisher","first-page":"1007","DOI":"10.1016\/j.ajo.2008.01.023","volume":"145","author":"GN Holland","year":"2008","unstructured":"Holland, G. N. et al. Analysis of recurrence patterns associated with toxoplasmic retinochoroiditis. Am. J. Ophthalmol. 145, 1007\u20131013. https:\/\/doi.org\/10.1016\/j.ajo.2008.01.023 (2008).","journal-title":"Am. J. Ophthalmol."},{"key":"43786_CR5","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetpar.2020.109195","volume":"286","author":"JP Dubey","year":"2020","unstructured":"Dubey, J. P., Murata, F. H. A., Cerqueira-C\u00e9zar, C. K., Kwok, O. C. H. & Su, C. Economic and public health importance of Toxoplasma gondii infections in sheep: 2009\u20132020. Vet. Parasitol. 286, 109195. https:\/\/doi.org\/10.1016\/j.vetpar.2020.109195 (2020).","journal-title":"Vet. Parasitol."},{"key":"43786_CR6","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetpar.2020.109185","volume":"288","author":"JP Dubey","year":"2020","unstructured":"Dubey, J. P. et al. All about Toxoplasma gondii infections in pigs: 2009\u20132020. Vet. Parasitol. 288, 109185. https:\/\/doi.org\/10.1016\/j.vetpar.2020.109185 (2020).","journal-title":"Vet. Parasitol."},{"key":"43786_CR7","doi-asserted-by":"publisher","first-page":"457","DOI":"10.4315\/0362-028X.JFP-14-328","volume":"78","author":"M Guo","year":"2015","unstructured":"Guo, M. et al. Prevalence and risk factors for Toxoplasma gondii infection in meat animals and meat products destined for human consumption. J. Food Prot. 78, 457\u2013476. https:\/\/doi.org\/10.4315\/0362-028X.JFP-14-328 (2015).","journal-title":"J. Food Prot."},{"key":"43786_CR8","first-page":"1","volume":"EN-995","author":"M Opsteegh","year":"2016","unstructured":"Opsteegh, M., Schares, G., Blaga, R. & van der Giessen, J. Experimental studies on Toxoplasma gondii in the main livestock species (GP\/EFSA\/BIOHAZ\/2013\/01). EFSA Support. Publ. EN-995, 1\u2013161 (2016).","journal-title":"EFSA Support. Publ."},{"key":"43786_CR9","volume":"16","author":"European Food Safety Authority (EFSA)","year":"2018","unstructured":"European Food Safety Authority (EFSA). Public health risks associated with food-borne parasites. EFSA J. 16, 5495 (2018).","journal-title":"EFSA J."},{"key":"43786_CR10","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0153856","volume":"11","author":"S Belluco","year":"2016","unstructured":"Belluco, S. et al. Investigating the determinants of Toxoplasma gondii prevalence in meat: A systematic review and meta-regression. PLoS One 11, e0153856. https:\/\/doi.org\/10.1371\/journal.pone.0153856 (2016).","journal-title":"PLoS One"},{"key":"43786_CR11","unstructured":"Centers for Disease Control and Prevention (CDC). Neglected parasitic infections in the United States: Toxoplasmosis. Preprint at (2016). https:\/\/www.cdc.gov\/parasites\/resources\/pdf\/npi_toxoplasmosis.pdf"},{"key":"43786_CR12","volume":"17","author":"A Thebault","year":"2021","unstructured":"Thebault, A., Kooh, P., Pilao-Cadavez, V. A., Gonzales-Barron, U. & Villena, I. Risk factors for sporadic toxoplasmosis: A systematic review and meta-analysis. Microb. Risk Anal. 17, 100133 (2021).","journal-title":"Microb. Risk Anal."},{"key":"43786_CR13","doi-asserted-by":"publisher","first-page":"2795","DOI":"10.1017\/S0950268814003185","volume":"143","author":"E Scallan","year":"2015","unstructured":"Scallan, E., Hoekstra, R. M., Mahon, B. E., Jones, T. F. & Griffin, P. M. An assessment of the human health impact of seven leading foodborne pathogens in the United States using disability adjusted life years. Epidemiol. Infect. 143, 2795\u20132804. https:\/\/doi.org\/10.1017\/S0950268814003185 (2015).","journal-title":"Epidemiol. Infect."},{"key":"43786_CR14","doi-asserted-by":"publisher","first-page":"322","DOI":"10.1089\/fpd.2019.2705","volume":"17","author":"S Monteiro Pires","year":"2020","unstructured":"Monteiro Pires, S., Jakobsen, L. S., Ellis-Iversen, J., Pessoa, J. & Ethelberg, S. Burden of disease estimates of seven pathogens commonly transmitted through foods in Denmark, 2017. Foodborne Pathog. Dis. 17, 322\u2013339. https:\/\/doi.org\/10.1089\/fpd.2019.2705 (2020).","journal-title":"Foodborne Pathog. Dis."},{"key":"43786_CR15","doi-asserted-by":"publisher","DOI":"10.1016\/j.parepi.2021.e00229","author":"J Marstrand","year":"2021","unstructured":"Marstrand, J., Kurtzhals, J. A. L., Fuchs, H. J., Nielsen, H. V. & Jokelainen, P. The disease burden of ocular toxoplasmosis in Denmark in 2019: Estimates based on laboratory testing of ocular samples and on publicly available register data. Parasite Epidemiol. Control https:\/\/doi.org\/10.1016\/j.parepi.2021.e00229 (2021).","journal-title":"Parasite Epidemiol. Control"},{"key":"43786_CR16","doi-asserted-by":"publisher","first-page":"875","DOI":"10.1016\/j.ijpara.2017.05.005","volume":"47","author":"I Gisbert Algaba","year":"2017","unstructured":"Gisbert Algaba, I. et al. A more sensitive, efficient and ISO 17025 validated magnetic capture real-time PCR method for the detection of archetypal Toxoplasma gondii strains in meat. Int. J. Parasitol. 47, 875\u2013884. https:\/\/doi.org\/10.1016\/j.ijpara.2017.05.005 (2017).","journal-title":"Int. J. Parasitol."},{"key":"43786_CR17","doi-asserted-by":"publisher","first-page":"193","DOI":"10.1016\/j.ijfoodmicro.2010.02.027","volume":"139","author":"M Opsteegh","year":"2010","unstructured":"Opsteegh, M. et al. Direct detection and genotyping of Toxoplasma gondii in meat samples using magnetic capture and PCR. Int. J. Food Microbiol. 139, 193\u2013201. https:\/\/doi.org\/10.1016\/j.ijfoodmicro.2010.02.027 (2010).","journal-title":"Int. J. Food Microbiol."},{"key":"43786_CR18","doi-asserted-by":"publisher","first-page":"107","DOI":"10.1016\/j.fm.2014.07.003","volume":"46","author":"M Gomez-Samblas","year":"2015","unstructured":"Gomez-Samblas, M., V\u00edlchez, S., Racero, J. C., Fuentes, M. V. & Osuna, A. Quantification and viability assays of Toxoplasma gondii in commercial \u201cSerrano\u201d ham samples using magnetic capture real-time qPCR and bioassay techniques. Food Microbiol. 46, 107\u2013113. https:\/\/doi.org\/10.1016\/j.fm.2014.07.003 (2015).","journal-title":"Food Microbiol."},{"key":"43786_CR19","doi-asserted-by":"publisher","DOI":"10.1016\/j.fawpar.2023.e00194","author":"F D\u00e1mek","year":"2023","unstructured":"D\u00e1mek, F. et al. Inactivation of Toxoplasma gondii in dry sausage and processed pork, and quantification of the pathogen in pig tissues prior to production. Food Waterborne Parasitol. https:\/\/doi.org\/10.1016\/j.fawpar.2023.e00194 (2023).","journal-title":"Food Waterborne Parasitol."},{"key":"43786_CR20","doi-asserted-by":"publisher","DOI":"10.1051\/parasite\/2015014","volume":"22","author":"A Aroussi","year":"2015","unstructured":"Aroussi, A. et al. Detection of Toxoplasma gondii DNA in horse meat from supermarkets in France and performance evaluation of two serological tests. Parasite 22, 14. https:\/\/doi.org\/10.1051\/parasite\/2015014 (2015).","journal-title":"Parasite"},{"key":"43786_CR21","doi-asserted-by":"publisher","first-page":"112","DOI":"10.1016\/j.fm.2016.04.005","volume":"58","author":"M Gomez-Samblas","year":"2016","unstructured":"Gomez-Samblas, M., V\u00edlchez, S., Racero, J. C., Fuentes, M. V. & Osuna, A. Toxoplasma gondii detection and viability assays in ham legs and shoulders from experimentally infected pigs. Food Microbiol. 58, 112\u2013120. https:\/\/doi.org\/10.1016\/j.fm.2016.04.005 (2016).","journal-title":"Food Microbiol."},{"key":"43786_CR22","doi-asserted-by":"publisher","first-page":"213","DOI":"10.1016\/j.fm.2017.02.010","volume":"65","author":"L Herrero","year":"2017","unstructured":"Herrero, L. et al. Toxoplasma gondii in raw and dry-cured ham: The influence of the curing process. Food Microbiol. 65, 213\u2013220. https:\/\/doi.org\/10.1016\/j.fm.2017.02.010 (2017).","journal-title":"Food Microbiol."},{"key":"43786_CR23","doi-asserted-by":"publisher","first-page":"497","DOI":"10.1016\/j.vetpar.2013.06.018","volume":"197","author":"J Jur\u00e1nkov\u00e1","year":"2013","unstructured":"Jur\u00e1nkov\u00e1, J. et al. Predilection sites of Toxoplasma gondii tissue cysts in sheep. Vet. Parasitol. 197, 497\u2013502. https:\/\/doi.org\/10.1016\/j.vetpar.2013.06.018 (2013).","journal-title":"Vet. Parasitol."},{"key":"43786_CR24","doi-asserted-by":"publisher","DOI":"10.1186\/s13567-015-0177-0","volume":"46","author":"A Burrells","year":"2015","unstructured":"Burrells, A. et al. Vaccination of pigs with the S48 strain of Toxoplasma gondii\u2014Safer meat for human consumption. Vet. Res. 46, 47. https:\/\/doi.org\/10.1186\/s13567-015-0177-0 (2015).","journal-title":"Vet. Res."},{"key":"43786_CR25","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2019.02223","volume":"10","author":"M Rahman","year":"2019","unstructured":"Rahman, M. et al. QuilA-adjuvanted Toxoplasma gondii lysate antigens trigger robust antibody and IFN\u03b3\u2009+\u2009T cell responses in pigs leading to reduction in parasite DNA in tissues upon challenge infection. Front. Immunol. 10, 2223. https:\/\/doi.org\/10.3389\/fimmu.2019.02223 (2019).","journal-title":"Front. Immunol."},{"key":"43786_CR26","doi-asserted-by":"publisher","DOI":"10.1186\/s13071-020-04275-z","volume":"13","author":"M Fern\u00e1ndez-Escobar","year":"2020","unstructured":"Fern\u00e1ndez-Escobar, M. et al. Isolation and genetic characterization of Toxoplasma gondii in Spanish sheep flocks. Parasit. Vectors 13, 396. https:\/\/doi.org\/10.1186\/s13071-020-04275-z (2020).","journal-title":"Parasit. Vectors"},{"key":"43786_CR27","doi-asserted-by":"publisher","DOI":"10.3389\/fvets.2020.604782","volume":"7","author":"M Fern\u00e1ndez-Escobar","year":"2020","unstructured":"Fern\u00e1ndez-Escobar, M. et al. Isolation, genotyping, and mouse virulence characterization of Toxoplasma gondii from free-ranging Iberian pigs. Front. Vet. Sci. 7, 604782. https:\/\/doi.org\/10.3389\/fvets.2020.604782 (2020).","journal-title":"Front. Vet. Sci."},{"key":"43786_CR28","doi-asserted-by":"publisher","first-page":"357","DOI":"10.1016\/j.ijpara.2020.04.001","volume":"50","author":"M Opsteegh","year":"2020","unstructured":"Opsteegh, M. et al. Methods to assess the effect of meat processing on viability of Toxoplasma gondii: Towards replacement of mouse bioassay by in vitro testing. Int. J. Parasitol. 50, 357\u2013369. https:\/\/doi.org\/10.1016\/j.ijpara.2020.04.001 (2020).","journal-title":"Int. J. Parasitol."},{"key":"43786_CR29","doi-asserted-by":"publisher","first-page":"1637","DOI":"10.1128\/EC.05182-11","volume":"10","author":"KR Buchholz","year":"2011","unstructured":"Buchholz, K. R. et al. Identification of tissue cyst wall components by transcriptome analysis of in vivo and in vitro Toxoplasma gondii bradyzoites. Eukaryot. Cell 10, 1637\u20131647. https:\/\/doi.org\/10.1128\/EC.05182-11 (2011).","journal-title":"Eukaryot. Cell"},{"key":"43786_CR30","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1002567","author":"AJ Reid","year":"2012","unstructured":"Reid, A. J. et al. Comparative genomics of the apicomplexan parasites Toxoplasma gondii\u00a0and Neospora caninum: Coccidia differing in host range and transmission strategy. PLoS Pathog. https:\/\/doi.org\/10.1371\/journal.ppat.1002567 (2012).","journal-title":"PLoS Pathog."},{"key":"43786_CR31","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2164-15-806","volume":"15","author":"KJ Pittman","year":"2014","unstructured":"Pittman, K. J., Aliota, M. T. & Knoll, L. J. Dual transcriptional profiling of mice and Toxoplasma gondii during acute and chronic infection. BMC Genomics 15, 806. https:\/\/doi.org\/10.1186\/1471-2164-15-806 (2014).","journal-title":"BMC Genomics"},{"key":"43786_CR32","doi-asserted-by":"publisher","DOI":"10.1186\/s12864-015-1225-x","volume":"16","author":"AB Hehl","year":"2015","unstructured":"Hehl, A. B. et al. Asexual expansion of Toxoplasma gondii merozoites is distinct from tachyzoites and entails expression of non-overlapping gene families to attach, invade, and replicate within feline enterocytes. BMC Genom. 16, 66. https:\/\/doi.org\/10.1186\/s12864-015-1225-x (2015).","journal-title":"BMC Genom."},{"key":"43786_CR33","doi-asserted-by":"publisher","DOI":"10.1186\/s12864-019-6213-0","volume":"20","author":"AL Garfoot","year":"2019","unstructured":"Garfoot, A. L., Wilson, G. M., Coon, J. J. & Knoll, L. J. Proteomic and transcriptomic analyses of early and late-chronic Toxoplasma gondii infection show novel and stage-specific transcripts. BMC Genomics 20, 859. https:\/\/doi.org\/10.1186\/s12864-019-6213-0 (2019).","journal-title":"BMC Genomics"},{"key":"43786_CR34","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-018-37671-8","volume":"9","author":"C Ramakrishnan","year":"2019","unstructured":"Ramakrishnan, C. et al. An experimental genetically attenuated live vaccine to prevent transmission of Toxoplasma gondii by cats. Sci. Rep. 9, 1474. https:\/\/doi.org\/10.1038\/s41598-018-37671-8 (2019).","journal-title":"Sci. Rep."},{"key":"43786_CR35","doi-asserted-by":"publisher","first-page":"359","DOI":"10.1016\/j.cell.2019.12.013","volume":"180","author":"BS Waldman","year":"2020","unstructured":"Waldman, B. S. et al. Identification of a master regulator of differentiation in Toxoplasma. Cell 180, 359-372e16. https:\/\/doi.org\/10.1016\/j.cell.2019.12.013 (2020).","journal-title":"Cell"},{"key":"43786_CR36","doi-asserted-by":"publisher","first-page":"491","DOI":"10.1016\/j.ijpara.2023.03.004","volume":"53","author":"A Colos-Arango","year":"2023","unstructured":"Colos-Arango, A., Largo-de la Torre, A., Calero-Bernal, R., Ortega-Mora, L. M. & Regidor-Cerrillo, J. Short-term culture adaptation of Toxoplasma gondii archetypal II and III field isolates affects cystogenic capabilities and modifies virulence in mice. Int. J. Parasitol. 53, 491\u2013504. https:\/\/doi.org\/10.1016\/j.ijpara.2023.03.004 (2023).","journal-title":"Int. J. Parasitol."},{"key":"43786_CR37","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2022.1021556","author":"A de la Largo- Torre","year":"2022","unstructured":"de la Largo- Torre, A. et al. Archetypal type II and III Toxoplasma gondii oocysts induce different immune responses and clinical outcomes in experimentally infected piglets. Front. Immunol. https:\/\/doi.org\/10.3389\/fimmu.2022.1021556 (2022).","journal-title":"Front. Immunol."},{"key":"43786_CR38","doi-asserted-by":"publisher","DOI":"10.1128\/aac.01448-24","author":"R S\u00e1nchez-S\u00e1nchez","year":"2025","unstructured":"S\u00e1nchez-S\u00e1nchez, R. et al. Treatment with BKI-1748 after Toxoplasma gondii systemic dissemination in experimentally infected pregnant sheep improves fetal and lamb mortality and morbidity and prevents congenital infection. Antimicrob. Agents Chemother. https:\/\/doi.org\/10.1128\/aac.01448-24 (2025).","journal-title":"Antimicrob. Agents Chemother."},{"key":"43786_CR39","volume-title":"Toxoplasmosis of animals and humans.","author":"JP Dubey","year":"2010","unstructured":"Dubey, J. P. Toxoplasmosis of animals and humans. 2nd ed. (CRC Press, 2010).","edition":"2nd ed"},{"key":"43786_CR40","doi-asserted-by":"publisher","first-page":"1651","DOI":"10.1017\/s003118200800509x","volume":"135","author":"J Regidor-Cerrillo","year":"2008","unstructured":"Regidor-Cerrillo, J. et al. Isolation and genetic characterization of Neospora caninum from asymptomatic calves in Spain. Parasitology 135, 1651\u20131659. https:\/\/doi.org\/10.1017\/s003118200800509x (2008).","journal-title":"Parasitology"},{"issue":"1","key":"43786_CR41","doi-asserted-by":"publisher","DOI":"10.1186\/s13567-021-00953-7","volume":"52","author":"M Fern\u00e1ndez-Escobar","year":"2021","unstructured":"Fern\u00e1ndez-Escobar, M. et al. In vivo and in vitro models show unexpected degrees of virulence among Toxoplasma gondii type II and III isolates from sheep. Vet. Res. 82. https:\/\/doi.org\/10.1186\/s13567-021-00953-7 (2021).","journal-title":"Vet. Res."},{"key":"43786_CR42","doi-asserted-by":"publisher","DOI":"10.1016\/j.fm.2021.103816","author":"M Kim","year":"2021","unstructured":"Kim, M. et al. Quantification of viable protozoan parasites on leafy greens using molecular methods. Food Microbiol. https:\/\/doi.org\/10.1016\/j.fm.2021.103816 (2021).","journal-title":"Food Microbiol."},{"key":"43786_CR43","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijfoodmicro.2023.110391","author":"M Kim","year":"2023","unstructured":"Kim, M. et al. Molecular detection and viability discrimination of zoonotic protozoan pathogens in oysters and seawater. Int. J. Food Microbiol. https:\/\/doi.org\/10.1016\/j.ijfoodmicro.2023.110391 (2023).","journal-title":"Int. J. Food Microbiol."},{"key":"43786_CR44","doi-asserted-by":"publisher","first-page":"1065","DOI":"10.1017\/S0031182008004599","volume":"135","author":"M Strohbusch","year":"2008","unstructured":"Strohbusch, M., M\u00fcller, N., Hemphill, A., Greif, G. & Gottstein, B. NcGRA2 as a molecular target to assess the parasiticidal activity of toltrazuril against Neospora caninum. Parasitology 135, 1065\u20131073. https:\/\/doi.org\/10.1017\/S0031182008004599 (2008).","journal-title":"Parasitology"},{"key":"43786_CR45","doi-asserted-by":"publisher","first-page":"64","DOI":"10.2174\/1874421400802010064","volume":"2","author":"M Strohbusch","year":"2008","unstructured":"Strohbusch, M., M\u00fcller, N., Hemphill, A., Greif, G. & Gottstein, B. NcGRA2-RT-PCR to detect live versus dead parasites in Neospora caninum-infected mice. Open. Parasitol. J. 2, 64\u201368 (2008).","journal-title":"Open. Parasitol. J."},{"key":"43786_CR46","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-023-38383-4","volume":"13","author":"D \u0110ermi\u0107","year":"2023","unstructured":"\u0110ermi\u0107, D. et al. Reverse transcription-quantitative PCR (RT-qPCR) without the need for prior removal of DNA. Sci. Rep. 11470. https:\/\/doi.org\/10.1038\/s41598-023-38383-4 (2023).","journal-title":"Sci. Rep."},{"key":"43786_CR47","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.02866-18","author":"LM Yeoh","year":"2019","unstructured":"Yeoh, L. M., Lee, V. V., McFadden, G. I. & Ralph, S. A. Alternative splicing in apicomplexan parasites. mBio https:\/\/doi.org\/10.1128\/mBio.02866-18 (2019).","journal-title":"mBio"},{"key":"43786_CR48","doi-asserted-by":"publisher","first-page":"555","DOI":"10.1016\/j.ijpara.2017.12.009","volume":"48","author":"I Gisbert Algaba","year":"2018","unstructured":"Gisbert Algaba, I. et al. Pork as a source of transmission of Toxoplasma gondii to humans: a parasite burden study in pig tissues after infection with different strains of Toxoplasma gondii as a function of time and different parasite stages. Int. J. Parasitol. 48, 555\u2013560. https:\/\/doi.org\/10.1016\/j.ijpara.2017.12.009 (2018).","journal-title":"Int. J. Parasitol."},{"key":"43786_CR49","doi-asserted-by":"publisher","first-page":"13","DOI":"10.1016\/j.vetpar.2019.04.004","volume":"269","author":"S Rani","year":"2019","unstructured":"Rani, S. et al. Toxoplasma gondii tissue cyst formation and density of tissue cysts in shoulders of pigs 7 and 14 days after feeding infected mice tissues. Vet. Parasitol. 269, 13\u201315. https:\/\/doi.org\/10.1016\/j.vetpar.2019.04.004 (2019).","journal-title":"Vet. Parasitol."},{"key":"43786_CR50","doi-asserted-by":"publisher","first-page":"47","DOI":"10.1016\/j.fm.2013.11.001","volume":"39","author":"H Neumayerov\u00e1","year":"2014","unstructured":"Neumayerov\u00e1, H. et al. Survival of experimentally induced Toxoplasma gondii tissue cysts in vacuum packed goat meat and dry fermented goat meat sausages. Food Microbiol. 39, 47\u201352. https:\/\/doi.org\/10.1016\/j.fm.2013.11.001 (2014).","journal-title":"Food Microbiol."}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-026-43786-0","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-026-43786-0.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-026-43786-0.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,5,5]],"date-time":"2026-05-05T23:43:38Z","timestamp":1778024618000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-026-43786-0"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,3,21]]},"references-count":50,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2026,12]]}},"alternative-id":["43786"],"URL":"https:\/\/doi.org\/10.1038\/s41598-026-43786-0","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,3,21]]},"assertion":[{"value":"28 July 2025","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"6 March 2026","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"21 March 2026","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"The authors declare no competing interests.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}},{"value":"All animal procedures were approved by the Animal Welfare Committee of the Community of Madrid, Spain (PROEX 293.7\/20 and PROEX 091.5\/24), following procedures described in Spanish and EU regulations (Law No. 3\/2007, R.D. 53\/2013, and Council Directive 2010\/63\/EU). The procedures were carried out at the registered experimental animal facility Department of Animal Health, Faculty of Veterinary Medicine, Complutense University of Madrid (user center registration code: ES280790000155).","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}}],"article-number":"14370"}}