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In the present study, we have determined in vitro growth rates of seven <jats:italic>T. pallidum<\/jats:italic> strains including three from the Nichols-like cluster (DAL-1, Madras, and Haiti B) and four from the SS14-like cluster (Mexico A, SS14, Grady, and Philadelphia 1). Despite the observed considerable variability, the results of standard subcultures identified growth differences between the two clusters during the long-term cultivation. Furthermore, in vitro monocultures with defined inoculum revealed differences among individual strains. During three week-long binary co-cultivations of seven strains (<jats:italic>n<\/jats:italic>\u2009=\u200921), different growth rates were confirmed for individual strains (<jats:italic>p<\/jats:italic>\u2009&lt;\u20090.001) using PCR amplicon sequencing of genomic regions differentiating treponemal-pairs. The order of strains by decreasing growth rate in vitro was DAL-1, Madras, Mexico A, Haiti B, SS14, Grady, and Philadelphia 1. While the generation time of strain DAL-1 was 32.97\u00a0h, the slowest strain, Philadelphia 1, had generation time 43.5\u00a0h. These experiments revealed significant physiological differences between the <jats:italic>T. pallidum<\/jats:italic> strains, which may also be involved in the variable presentation of syphilis symptoms observed in previous decades.<\/jats:p>","DOI":"10.1038\/s41598-025-18827-9","type":"journal-article","created":{"date-parts":[[2025,9,26]],"date-time":"2025-09-26T11:40:09Z","timestamp":1758886809000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["In vitro analysis of seven syphilis-causing Treponema pallidum strains revealed inherent growth rate differences"],"prefix":"10.1038","volume":"15","author":[{"given":"Juraj","family":"Bos\u00e1k","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mat\u011bj","family":"Hrala","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Petr","family":"Andrla","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eli\u0161ka","family":"Vrbov\u00e1","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Petra","family":"Posp\u00ed\u0161ilov\u00e1","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David","family":"\u0160majs","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,9,26]]},"reference":[{"key":"18827_CR1","doi-asserted-by":"publisher","first-page":"e0143304","DOI":"10.1371\/journal.pone.0143304","volume":"10","author":"L Newman","year":"2015","unstructured":"Newman, L. et al. 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