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<p>Extracellular vesicles (EVs) have emerged as important mediators of bacterial interspecies interactions and may play critical roles in food spoilage dynamics, but co-isolated contaminants hinder their accurate characterization. To address this, we compared conventional ultracentrifugation (UC) and two combined methods for isolating A<i>eromonas salmonicida</i> EVs (ASEVs). The ultrafiltration–UC–density gradient centrifugation (FCD) method achieved a 6.9-fold purity increase over UC. Comparative proteomics revealed that contaminants mainly originated from cytoplasm, and flagellum-, pili-, and phage-associated proteins were identified as purity markers. The ASEVs proteins are primarily located in membrane structures and mainly implicated in metabolic and membrane transport pathways. Functional assays demonstrated that ASEVs inhibited planktonic growth of <i>Pseudomonas fluorescens</i> and reduced biofilm formation by 40.48%. This study establishes a theoretical foundation for elucidating the characteristics of food spoilage bacteria EVs and their roles in bacterial interspecies interactions.</p>

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Characterization of high-purity Aeromonas salmonicida extracellular vesicles and their inhibitory activity against Pseudomonas fluorescens biofilm formation

  • 期刊论文

  • 发表日期：

    2026年6月2日

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  • Zhenzhen Ning,
  • Wei Rao,
  • Liangting Shao,
  • Yang Dong,
  • Silu Liu,
  • Xinqi Yang,
  • Xinglian Xu,
  • Huhu Wang

摘要

Extracellular vesicles (EVs) have emerged as important mediators of bacterial interspecies interactions and may play critical roles in food spoilage dynamics, but co-isolated contaminants hinder their accurate characterization. To address this, we compared conventional ultracentrifugation (UC) and two combined methods for isolating Aeromonas salmonicida EVs (ASEVs). The ultrafiltration–UC–density gradient centrifugation (FCD) method achieved a 6.9-fold purity increase over UC. Comparative proteomics revealed that contaminants mainly originated from cytoplasm, and flagellum-, pili-, and phage-associated proteins were identified as purity markers. The ASEVs proteins are primarily located in membrane structures and mainly implicated in metabolic and membrane transport pathways. Functional assays demonstrated that ASEVs inhibited planktonic growth of Pseudomonas fluorescens and reduced biofilm formation by 40.48%. This study establishes a theoretical foundation for elucidating the characteristics of food spoilage bacteria EVs and their roles in bacterial interspecies interactions.