{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T01:27:51Z","timestamp":1760059671110,"version":"build-2065373602"},"reference-count":27,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2025,6,29]],"date-time":"2025-06-29T00:00:00Z","timestamp":1751155200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Open Access Publishing Fund of Leipzig University"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Toxoplasma gondii, an obligate intracellular protozoan, poses significant risks to public health due to its widespread distribution and potential for severe congenital and neurological complications. The fast-replicating tachyzoite stage is crucial for acute infection and laboratory studies, yet effective inactivation methods remain inadequately explored. This study evaluates various chemical and physical approaches to inactivate T. gondii tachyzoites in vitro. Using a combination of GFP fluorescence and viability assays, we demonstrated the complete inactivation of tachyzoites with ethanol (\u226530%), hydrogen peroxide (\u22653%), o-hydroxydiphenyl fatty acid eutectic with peracetic acid (\u22651%), and heat treatment at 60 \u00b0C for 30 min. Our findings highlight the importance of concentration, solvent choice, and exposure time in disinfection efficacy, providing a framework for improving laboratory safety protocols. These results contribute to the refinement of inactivation strategies, supporting safer handling and research on T. gondii in vitro while reducing reliance on animal models.<\/jats:p>","DOI":"10.3390\/microorganisms13071517","type":"journal-article","created":{"date-parts":[[2025,6,30]],"date-time":"2025-06-30T03:54:28Z","timestamp":1751255668000},"page":"1517","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Low Tenacity of Toxoplasma\u00a0gondii Tachyzoites In Vitro"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7836-6446","authenticated-orcid":false,"given":"Thomas","family":"Grochow","sequence":"first","affiliation":[{"name":"Institute of Veterinary Anatomy, Histology and Embryology, Faculty of Veterinary Medicine, Leipzig University, 04103 Leipzig, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mirjam","family":"Kalusa","sequence":"additional","affiliation":[{"name":"Institute of Veterinary Anatomy, Histology and Embryology, Faculty of Veterinary Medicine, Leipzig University, 04103 Leipzig, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pauline","family":"Tonndorf-Martini","sequence":"additional","affiliation":[{"name":"Institute of Veterinary Anatomy, Histology and Embryology, Faculty of Veterinary Medicine, Leipzig University, 04103 Leipzig, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7974-9281","authenticated-orcid":false,"given":"Nicole","family":"R\u00f6hrmann","sequence":"additional","affiliation":[{"name":"Institute of Veterinary Anatomy, Histology and Embryology, Faculty of Veterinary Medicine, Leipzig University, 04103 Leipzig, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Simone A.","family":"Fietz","sequence":"additional","affiliation":[{"name":"Institute of Veterinary Anatomy, Histology and Embryology, Faculty of Veterinary Medicine, Leipzig University, 04103 Leipzig, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lea-Christina","family":"Murnik","sequence":"additional","affiliation":[{"name":"Institute of Parasitology, Faculty of Veterinary Medicine, Leipzig University, 04103 Leipzig, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,6,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1217","DOI":"10.1016\/S0020-7519(00)00124-7","article-title":"Toxoplasma gondii: From animals to humans","volume":"30","author":"Tenter","year":"2000","journal-title":"Int. J. Parasitol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"463","DOI":"10.1146\/annurev-micro-091014-104100","article-title":"The lytic cycle of Toxoplasma gondii: 15 years later","volume":"69","author":"Blader","year":"2015","journal-title":"Annu. Rev. Microbiol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"636","DOI":"10.1084\/jem.132.4.636","article-title":"The Toxoplasma gondii oocyst from cat feces","volume":"132","author":"Dubey","year":"1970","journal-title":"J. Exp. Med."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"928","DOI":"10.2307\/3286588","article-title":"Oral infections with Toxoplasma cysts and oocysts in felines, other mammals, and in birds","volume":"58","author":"Miller","year":"1972","journal-title":"J. Parasitol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"267","DOI":"10.1128\/CMR.11.2.267","article-title":"Structures of Toxoplasma gondii tachyzoites, bradyzoites, and sporozoites and biology and development of tissue cysts","volume":"11","author":"Dubey","year":"1998","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_6","unstructured":"Dubey, J.P. (2010). Toxoplasmosis of Animals and Humans, CRC Press. [2nd ed.]."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1965","DOI":"10.1016\/S0140-6736(04)16412-X","article-title":"Toxoplasmosis","volume":"363","author":"Montoya","year":"2004","journal-title":"Lancet"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Bigna, J.J., Tochie, J.N., Tounouga, D.N., Bekolo, A.O., Ymele, N.S., Youda, E.L., Sime, P.S., and Nansseu, J.R. (2020). Global, regional, and country seroprevalence of Toxoplasma gondii in pregnant women: A systematic review, modelling and meta-analysis. Sci. Rep., 10.","DOI":"10.1038\/s41598-020-69078-9"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1242\/jeb.074153","article-title":"Toxoplasma gondii infection and behaviour\u2014location, location, location?","volume":"216","author":"McConkey","year":"2013","journal-title":"J. Exp. Biol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1111\/pim.12157","article-title":"Toxoplasma gondii induced neuronal alterations","volume":"37","author":"Parlog","year":"2015","journal-title":"Parasite Immunol."},{"key":"ref_11","first-page":"81","article-title":"Cyst formation from virulent RH strain of Toxoplasma gondii tachyzoite: In vitro cultivation","volume":"11","author":"Salimi","year":"2016","journal-title":"Iran. J. Parasitol."},{"key":"ref_12","first-page":"145","article-title":"In vitro culture of Toxoplasma gondii in HeLa, Vero, RBK and A549 cell lines","volume":"26","author":"Jabari","year":"2018","journal-title":"Infez. Med."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"e026420","DOI":"10.1590\/s1984-29612021040","article-title":"Techniques for inactivating Toxoplasma gondii oocysts: A systematic review","volume":"30","author":"Paschoal","year":"2021","journal-title":"Rev. Bras. Parasitol. Vet."},{"key":"ref_14","unstructured":"Deutsche Veterin\u00e4rmedizinische Gesellschaft, e.V. (2025, January 31). DVG-Desinfektionsmittelliste f\u00fcr den Tierhaltungsbereich. Available online: https:\/\/www.desinfektion-dvg.de\/dvg-desinfektionsmittellisten\/tierhaltung."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"306","DOI":"10.1080\/20477724.2018.1514137","article-title":"A review on inactivation methods of Toxoplasma gondii in foods","volume":"112","author":"Alizadeh","year":"2018","journal-title":"Pathog. Glob. Health."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Montazeri, M., Mehrzadi, S., Sharif, M., Sarvi, S., Tanzifi, A., Aghayan, S.A., and Daryani, A. (2018). Drug resistance in Toxoplasma gondii. Front. Microbiol., 9.","DOI":"10.3389\/fmicb.2018.02587"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Hua, Q.-Q., Lin, X.-J., Xiang, S.-P., Jiang, L.-Y., Cai, J.-H., Sun, J.-M., Tan, F., and Mou, Y.-N. (2023). Two small-molecule inhibitors of Toxoplasma gondii proliferation in vitro. Front. Cell. Infect. Microbiol., 13.","DOI":"10.3389\/fcimb.2023.1145824"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"126","DOI":"10.1093\/infdis\/jiaa072","article-title":"A sialic acid-binding protein SABP1 of Toxoplasma gondii mediates host cell attachment and invasion","volume":"222","author":"Xing","year":"2020","journal-title":"J. Infect. Dis."},{"key":"ref_19","first-page":"152","article-title":"The survival of Toxoplasma gondii trophozoites in changes in osmotic pressure","volume":"54","author":"Saari","year":"1976","journal-title":"Med. Biol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1186\/1756-3305-1-35","article-title":"Efficacy of common laboratory disinfectants and heat on killing trypanosomatid parasites","volume":"1","author":"Wang","year":"2008","journal-title":"Parasit. Vectors."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Alves, E., Benns, H.J., Magnus, L., Dominicus, C., Dobai, T., Blight, J., Wincott, C.J., and Child, M.A. (2021). An extracellular redox signal triggers calcium release and impacts the asexual development of Toxoplasma gondii. Front. Cell. Infect. Microbiol., 11.","DOI":"10.3389\/fcimb.2021.728425"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1016\/j.vetpar.2017.07.018","article-title":"Improvement of in vitro evaluation of chemical disinfectants for efficacy on Cryptosporidium parvum oocysts","volume":"245","author":"Delling","year":"2017","journal-title":"Vet. Parasitol."},{"key":"ref_23","first-page":"e20170257","article-title":"Hydrogen peroxide and chlorine dioxide against parasite Ichthyophthirius multifiliis (Protozoa, Ciliophora) in jundi\u00e1 fingerlings","volume":"47","author":"Silva","year":"2017","journal-title":"Cienc. Rural"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Lee, M., and Lee, E. (2007). The effectiveness of hydrogen peroxide liquid or gas plasma on protozoan oocysts. Can. J. Infect. Dis. Med. Microbiol., 18.","DOI":"10.1155\/2007\/265870"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"201","DOI":"10.2307\/3283016","article-title":"Effect of high temperature on infectivity of Toxoplasma gondii tissue cysts in pork","volume":"76","author":"Dubey","year":"1990","journal-title":"J. Parasitol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"687","DOI":"10.1089\/fpd.2007.0060","article-title":"Methods for inactivation of Toxoplasma gondii cysts in meat and tissues of experimentally infected sheep","volume":"5","author":"Tawfik","year":"2008","journal-title":"Foodborne Pathog. Dis."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"496","DOI":"10.20506\/rst.16.2.1032","article-title":"Parasites associated with pork and pork products","volume":"16","author":"Gamble","year":"1997","journal-title":"Rev. Sci. Tech."}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/13\/7\/1517\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:01:10Z","timestamp":1760032870000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/13\/7\/1517"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,6,29]]},"references-count":27,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2025,7]]}},"alternative-id":["microorganisms13071517"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms13071517","relation":{},"ISSN":["2076-2607"],"issn-type":[{"type":"electronic","value":"2076-2607"}],"subject":[],"published":{"date-parts":[[2025,6,29]]}}}