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10.3390/microorganisms13122667
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Article

Lacticaseibacillus casei Combats Biofilm Formation and Exhibits Antibacterial Activity Against Clinical Isolates of Staphylococcus aureus, Salmonella enterica, and Escherichia coli

by
Despoina Eugenia Kiousi
1,
Sotiris Kyriakou
2,
Christos Efstathiou
1,
Stylianos Didaskalou
1,
Maria Koffa
1,
Aglaia Pappa
1,
Maria Panopoulou
3,
Mihalis I. Panayiotidis
2,4 and
Alex Galanis
1,*
1
Department of Molecular Biology and Genetics, Faculty of Health Sciences, Democritus University of Thrace, 68100 Alexandroupolis, Greece
2
Department of Cancer Genetics, Therapeutics and Ultrastructural Pathology, The Cyprus Institute of Neurology and Genetics, 2371 Nicosia, Cyprus
3
Department of Medicine, Faculty of Health Sciences, Democritus University of Thrace, 68100 Alexandroupolis, Greece
4
Department of Comparative Biomedical Sciences, School of Veterinary Medicine, Mississippi State University, Starkville, MS 39762, USA
*
Author to whom correspondence should be addressed.
Microorganisms 2025, 13(12), 2667; https://doi.org/10.3390/microorganisms13122667
Submission received: 19 October 2025 / Revised: 17 November 2025 / Accepted: 19 November 2025 / Published: 24 November 2025
(This article belongs to the Special Issue Advanced Research on Microbial Biofilms—a Themed Issue in Honor of Professor Gianfranco Donelli)
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Abstract

Biofilm-forming pathogens are a major cause of persistent infections, showing limited response to antibiotic treatment. The search for alternative strategies has therefore driven extensive research into the antimicrobial potential of beneficial microorganisms. In the present study, the antibacterial and antibiofilm activity of the commercial probiotic strain Lacticaseibacillus casei ATCC 393 (Lc393) was examined against clinical isolates of Staphylococcus aureus, Salmonella enterica subsp. enterica serovar Enteritidis and Escherichia coli. Lc393 reduced pathogen viability and attachment to the colon adenocarcinoma cell line HT-29, with maximal effects recorded against S. aureus. Confocal microscopy visualization of the lactobacilli-pathogens-host interface revealed that Lc393 binds loosely to both host cells and pathogens. The Lc393 cell-free culture supernatant (CFCS) significantly reduced planktonic growth, biofilm mass, and viability of cells in biofilm (>2 logCFU reduction, p < 0.05) and downregulated genes involved in the early stages of biofilm formation in S. aureus (i.e., icaA, fnbpA, eno). In silico analysis of the Lc393 genome identified two bacteriocin clusters, along with genes related to ethanol and organic acid production. Based on in silico predictions and a bacteriocin zymogram, the strain cannot produce functional antimicrobial peptides. Untargeted metabolomics based on UPLC/MS further revealed the presence of putative antimicrobial metabolites. Collectively, our findings highlight the antimicrobial potential of Lc. casei ATCC 393 and support its further investigation for combating clinically relevant human pathogens.
Keywords: Lacticaseibacillus casei; antimicrobial; antibiofilm; pathogens; untargeted metabolomics; cell-free culture supernatants; probiotics Lacticaseibacillus casei; antimicrobial; antibiofilm; pathogens; untargeted metabolomics; cell-free culture supernatants; probiotics

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MDPI and ACS Style

Kiousi, D.E.; Kyriakou, S.; Efstathiou, C.; Didaskalou, S.; Koffa, M.; Pappa, A.; Panopoulou, M.; Panayiotidis, M.I.; Galanis, A. Lacticaseibacillus casei Combats Biofilm Formation and Exhibits Antibacterial Activity Against Clinical Isolates of Staphylococcus aureus, Salmonella enterica, and Escherichia coli. Microorganisms 2025, 13, 2667. https://doi.org/10.3390/microorganisms13122667

AMA Style

Kiousi DE, Kyriakou S, Efstathiou C, Didaskalou S, Koffa M, Pappa A, Panopoulou M, Panayiotidis MI, Galanis A. Lacticaseibacillus casei Combats Biofilm Formation and Exhibits Antibacterial Activity Against Clinical Isolates of Staphylococcus aureus, Salmonella enterica, and Escherichia coli. Microorganisms. 2025; 13(12):2667. https://doi.org/10.3390/microorganisms13122667

Chicago/Turabian Style

Kiousi, Despoina Eugenia, Sotiris Kyriakou, Christos Efstathiou, Stylianos Didaskalou, Maria Koffa, Aglaia Pappa, Maria Panopoulou, Mihalis I. Panayiotidis, and Alex Galanis. 2025. "Lacticaseibacillus casei Combats Biofilm Formation and Exhibits Antibacterial Activity Against Clinical Isolates of Staphylococcus aureus, Salmonella enterica, and Escherichia coli" Microorganisms 13, no. 12: 2667. https://doi.org/10.3390/microorganisms13122667

APA Style

Kiousi, D. E., Kyriakou, S., Efstathiou, C., Didaskalou, S., Koffa, M., Pappa, A., Panopoulou, M., Panayiotidis, M. I., & Galanis, A. (2025). Lacticaseibacillus casei Combats Biofilm Formation and Exhibits Antibacterial Activity Against Clinical Isolates of Staphylococcus aureus, Salmonella enterica, and Escherichia coli. Microorganisms, 13(12), 2667. https://doi.org/10.3390/microorganisms13122667

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