20.500.12556/RUL-182930 Lactococcus cremoris decreases virulence‑associated properties, and modifies the protein profile of foodborne pathogen Campylobacter jejuni in biofilm Campylobacter jejuni, the most common foodborne pathogen, relies on adhesion, invasion, and biofilm formation for successful virulence and persistence in the food-processing environments. In this study, we investigated the effects of the transient bacterium with probiotic properties Lactococcus cremoris on C. jejuni virulence via an adhesion and invasion assay using human colon adenocarcinoma Caco-2 cells, culturability via colony-forming units, and proteomic adaptation in co-culture via label-free quantitative mass spectrometry. Caco-2 cell adhesion and invasion assay revealed that L. cremoris decreases C. jejuni adhesion (by 50%) and invasion (by 92%), particularly when inoculated prior to the pathogen. In mature (48 h old) dual-species biofilms, C. jejuni culturability decreased by 88%, while L. cremoris culturability remained unaffected, compared to single-species biofilms. Proteomic analysis of biofilm cells showed metabolic reprogramming in C. jejuni, with increased levels of proteins related to the TCA cycle, stress response, amino acid and nucleotide metabolism, chemotaxis, and energy production. The most upregulated proteins included the tungsten transporter TupA, LuxS, Dps, and hydrogenase subunits HydAB, indicating adaptations to nutrient limitation and oxidative stress. Nevertheless, the decreased culturability of C. jejuni suggests it is overwhelmed under the competitive conditions of dual-species biofilms. Our findings demonstrate that L. cremoris exerts strong antagonistic effects on C. jejuni, restricting its virulence, and triggering significant metabolic shifts in dual-species biofilms. These results highlight the potential of L. cremoris as a preventative strategy against C. jejuni adhesion, colonisation, and transmission. Campylobacter jejuni Lactococcus cremoris virulence adhesion biofilm quantitative proteomics stress response Campylobacter jejuni Lactococcus cremoris adhezija biofilm odziv na stres true false true Angleški jezik Slovenski jezik Članek v reviji 2026-05-28 13:51:44 2026-05-28 13:51:52 2026-05-29 04:37:02 0000-00-00 00:00:00 2026 0 0 13 str. art. 41 Vol. 18 2026 0000-00-00 Zaloznikova Objavljeno apc 0000-00-00 0000-00-00 0000-00-00 579.22/.26 1757-4749 10.1186/s13099-026-00827-3 275947267 RAZ_Jug_Blaz_2026.pdf RAZ_Jug_Blaz_2026.pdf 1 902B9EED7F8D7DBAA2BA3A3D330D7448 ba0ada6a17ea7d367679e9aae4045fef01d5eb91bbf0794dab5043b0fb2583ea 5257bc3f-5a8d-11f1-b0ab-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=234616 https://link.springer.com/article/10.1186/s13099-026-00827-3 1 4a2622d0-5a8b-11f1-b0ab-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=234615 Biotehniška fakulteta 0 0 0