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bioRxiv · 10.1101/2025.08.10.669548

Profiling of Burkholderia pseudomallei variants derived from Queensland clinical isolates

Coulon, P. M. L.·Ramsay, K. A.·Lee, A.·Ritmejeryte, E.·Pitt, M. E.·To, J.·Mediati, D. G.·Reed, S.·Harris, P. N. A.·Myers, G. S. A.
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Abstract

Burkholderia pseudomallei (Bp), an environmental bacterium and opportunistic pathogen endemic to tropical regions, is highly adaptive and thrives in diverse environments, from soil to human hosts. Bacterial adaptation is critical for survival, virulence modulation, and persistence during infection and can manifest as colony morphotype variation (CMV). While Bp adaptation has been well studied, CMV remains poorly understood. Here, we characterized five clinical Bp isolates exhibiting heterogeneous populations with rough and smooth colony morphologies. We used phenotypic assays, whole-genome sequencing, and proteomics to investigate the molecular pathways impacted by CMV - by comparing smooth and rough morphotypes. While phenotypic differences in protease activity, haemolysis, mucoid, iron uptake and antibiotic sensitivity --including to antimicrobial agents commonly used to treat infections--were rare, these traits alone could not distinguish morphotypes or group of isolates. Genomic comparisons revealed either no differences or limited isolate-specific mutations, which does not explain the overall difference in phenotypes. In contrast, proteomic analysis uncovered consistent shifts in protein abundance related to virulence, including quorum sensing, DNA methylation, and secretion systems. Rough variants showed higher expression of EPS-associated proteins, the BpsI3/R3 quorum sensing system, and the global regulator ScmR, whereas smooth variants upregulated type III/VI secretion and siderophore biosynthesis pathways. These findings suggest that CMV is driven by phase variation and regulatory mechanisms rather than punctual genomic modifications. Our study underscores the limitations of phenotype or genome-based classification alone in the context of CMV and highlights the value of integrated multi-omics approaches to uncover CMV-associated biomarkers, with potential applications in diagnostics and the development of targeted therapies against persistent and drug-resistant Bp infections.

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Coulon, P. M. L., Ramsay, K. A., Lee, A., Ritmejeryte, E., Pitt, M. E., To, J., Mediati, D. G., Reed, S., Harris, P. N. A., Myers, G. S. A.. 2025-08-11. Profiling of Burkholderia pseudomallei variants derived from Queensland clinical isolates. https://doi.org/10.1101/2025.08.10.669548

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