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Biofilms play a significant role in bacterial persistence and contribute to the development of chronic lesions. A recent study has shown that polymicrobial interactions between species are an important factor in biofilm formation, yet the precise mechanism of biofilm formation in\n                    <jats:italic>M. bovis<\/jats:italic>\n                    remains unknown. By assuming multiple pathogen infections in the bovine respiratory disease complex (BRDC), this study examined the characterisation of the polymicrobial relationship between\n                    <jats:italic>M. bovis<\/jats:italic>\n                    and\n                    <jats:italic>Trueperella pyogenes<\/jats:italic>\n                    (\n                    <jats:italic>T. pyogenes<\/jats:italic>\n                    ) during biofilm formation. Autopsies were performed on four Holstein calves (two chronic\n                    <jats:italic>Mycoplasma<\/jats:italic>\n                    pneumonia calves and two control calves). Bacterium-like aggregation structures (&gt;\u200910\u00a0\u03bcm), which were assumed to be biofilms of\n                    <jats:italic>M. bovis<\/jats:italic>\n                    in vivo, were observed adhering to the cilia in calves with\n                    <jats:italic>Mycoplasma<\/jats:italic>\n                    pneumonia.\n                    <jats:italic>M. bovis<\/jats:italic>\n                    released an extracellular matrix to connect with neighbouring bacteria and form a mature biofilm on the plate. Biofilm formation in the co-culture of\n                    <jats:italic>M. bovis<\/jats:italic>\n                    and\n                    <jats:italic>T. pyogenes<\/jats:italic>\n                    (strain T1: 1\u2009\u00d7\u200910\n                    <jats:sup>5<\/jats:sup>\n                    and 1\u2009\u00d7\u200910\n                    <jats:sup>6<\/jats:sup>\n                    \u00a0CFU\/well) significantly increased (\n                    <jats:italic>p<\/jats:italic>\n                    \u2009&lt;\u20090.05 and\n                    <jats:italic>p<\/jats:italic>\n                    \u2009&lt;\u20090.01; 64.1% and 64.8% increase) compared to that in a single culture of these bacteria. Furthermore, some large aggregates (&gt;\u200940\u00a0\u03bcm), composed of\n                    <jats:italic>M. bovis<\/jats:italic>\n                    and\n                    <jats:italic>T. pyogenes<\/jats:italic>\n                    , were observed. The morphological characteristics of this biofilm were similar to those observed in vivo compared to a single culture. In conclusion, the polymicrobial interaction between\n                    <jats:italic>M. bovis<\/jats:italic>\n                    and\n                    <jats:italic>T. pyogenes<\/jats:italic>\n                    induces biofilm formation, which is associated with increased resistance to antimicrobial agents, and this exacerbates the progression of chronic\n                    <jats:italic>Mycoplasma<\/jats:italic>\n                    pneumonia.\n                  <\/jats:p>","DOI":"10.1186\/s13567-025-01468-1","type":"journal-article","created":{"date-parts":[[2025,1,30]],"date-time":"2025-01-30T10:31:26Z","timestamp":1738233086000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Biofilm characterisation of Mycoplasma bovis co-cultured with Trueperella pyogenes"],"prefix":"10.1186","volume":"56","author":[{"given":"Koji","family":"Nishi","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9745-7792","authenticated-orcid":false,"given":"Satoshi","family":"Gondaira","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuki","family":"Hirano","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Masahide","family":"Ohashi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ayano","family":"Sato","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kazuya","family":"Matsuda","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tomohito","family":"Iwasaki","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takuya","family":"Kanda","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ryoko","family":"Uemura","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hidetoshi","family":"Higuchi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,1,30]]},"reference":[{"key":"1468_CR1","doi-asserted-by":"publisher","first-page":"225","DOI":"10.1016\/j.cvfa.2012.03.007","volume":"28","author":"LK Fox","year":"2012","unstructured":"Fox LK (2012) Mycoplasma mastitis: causes, transmission, and control. 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