Antibiofilm Power of Basil Essential Oil Against Fish-Originated Multidrug-Resistant Salmonella and Bacillus spp.: Targeting Biofilms on Food Contact Surfaces.

Abstract

The antimicrobial and antibiofilm efficacy of two Ocimum basilicum L., essential oils sourced from Colombia (BEOC) and Italy (BEOI), was evaluated against multidrug-resistant fish isolates of Salmonella enterica subsp. salamae, Bacillus thuringiensis, and Bacillus oceanisediminis-species for which such activity has not been previously reported. Using a fish-based model system (FBMS), we found that BEOI, rich in linalool (69.86%), exhibited stronger antimicrobial activity than camphor-dominated BEOC (24.61%). The antimicrobial effects of both EOs were strain- and concentration-dependent, with minimum bactericidal concentration (MBC) 3.75-15.0 µL/mL for BEOI and 15.0-30.0 µL/mL for BEOC. Pure linalool showed even greater potency (MBC: 0.0125 to 0.025 µL/mL). Confocal laser scanning microscopy revealed that BEOI induced severe membrane damage (27% of the cells within 1 h), ultimately leading to the death of 96% of the cells after 24 h. Biofilm formation, assessed in both FBMS and tryptone soy broth (TSB), was strain-dependent, with FBMS promoting higher biofilm production than TSB. Moreover, significant differences in biofilm morphotypes were observed, with the morphotype PDAR (pink dry and rough), characterized by only cellulose, being the most frequently exhibited by the strains (7/15), while BDAR (brown dry and rough), characterized by only curli, was the least expressed (7/15); the remaining strains presented morphotype RDAR. In addition, the strains in polystyrene surfaces accumulated more biomass than stainless steel 304. Notably, BEOI and linaool significantly reduced biofilm formation across all strains, with a reduction of 90% in S. enterica subsp. salamae strains (TJC19 and TJC21. These strains with the RDAR phenotype likely contribute to their strong biofilm-forming capacity. Our findings highlight BEOI's potential as a natural anti-biofilm agent in food processing environments, offering a promising strategy to combat multidrug-resistant bacteria biofilm-related challenges in the food industry.