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Studies have compared the currently dominant single nucleotide polymorphism (SNP) cluster I (pertussis toxin promoter allele,\n                      <jats:italic toggle=\"yes\">ptxP3<\/jats:italic>\n                      ) and previously dominant SNP cluster II (\n                      <jats:italic toggle=\"yes\">ptxP1<\/jats:italic>\n                      ) strains as planktonic cells. Since biofilm formation is linked with\n                      <jats:italic toggle=\"yes\">B. pertussis<\/jats:italic>\n                      pathogenesis\n                      <jats:italic toggle=\"yes\">in vivo<\/jats:italic>\n                      , this study compared the biofilm formation capabilities of representative strains of cluster I and cluster II. Confocal laser scanning microscopy found that the cluster I strain had a denser biofilm structure compared to the cluster II strain. Differences in protein abundance of the biofilm cells were then compared using tandem mass tagging and high-resolution multiple reaction monitoring. In total, 1,453 proteins were identified, of which 40 proteins had significant differential abundance between the two strains in biofilm conditions. Of particular interest was a large increase in the abundance of energy metabolism proteins (cytochrome proteins PetABC and BP3650) in the cluster I strain. When the abundance of these proteins was compared between six additional strains from each cluster, it was found that the protein abundance varied between all strains. These findings suggest that there are large levels of individual proteomic diversity between\n                      <jats:italic toggle=\"yes\">B. pertussis<\/jats:italic>\n                      strains in biofilm conditions despite the highly conserved genome of the species. Overall, this study revealed visual differences in biofilm structure between\n                      <jats:italic toggle=\"yes\">B. pertussis<\/jats:italic>\n                      strains and highlighted strain-specific variation in protein abundance that dominates potential cluster-specific changes that may be linked with the dominance of cluster I strains.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>\n                        <jats:italic toggle=\"yes\">Bordetella pertussis<\/jats:italic>\n                        causes whooping cough. The currently circulating cluster I strains have taken over previously dominant cluster II strains. It is important to understand the reasons behind this evolution to develop new strategies against the pathogen. Recent studies have shown that\n                        <jats:italic toggle=\"yes\">B. pertussis<\/jats:italic>\n                        can form biofilms during infection. This study compared the biofilm formation capabilities of a cluster I and a cluster II strain and identified visual differences in the biofilms. The protein abundance between these strains grown in biofilms was compared, and proteins identified with varied abundance were measured with additional strains from each cluster. It was found that despite the highly conserved genetics of the species, there was varied protein abundance between the additional strains. This study highlights that strain-specific variation in protein abundance during biofilm conditions may dominate the cluster-specific changes that may be linked to the dominance of cluster I strains.\n                      <\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/spectrum.01715-25","type":"journal-article","created":{"date-parts":[[2025,9,30]],"date-time":"2025-09-30T12:06:51Z","timestamp":1759234011000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":1,"title":["Proteomic comparison of epidemic Australian\n                    <i>Bordetella pertussis<\/i>\n                    biofilm cells"],"prefix":"10.1128","volume":"13","author":[{"given":"Hiroki","family":"Suyama","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/03r8z3t63","id-type":"ROR","asserted-by":"publisher"}],"name":"School of Biotechnology and Biomolecular Sciences, University of New 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