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Exploring the Activity of Bacteriocins Against Bacterial Biofilms Using a Simple and Inexpensive Assay
Matilde Fernández1,2, María Isabel Torices3, Claudia Teso3
1Department of Microbiology, Faculty of Sciences, C. Fuentenueva S/N, University of Granada, Granada, Spain. matildefernandez@ugr.es.
Probiotics and Antimicrobial Proteins
|September 26, 2025
Summary
A new method for growing bacterial biofilms on glass beads in microtiter plates is presented. This approach effectively tests antimicrobial susceptibility, highlighting bacteriocins like nisin and AS-48 as promising agents against resistant biofilms.
Area of Science:
- Microbiology
- Antimicrobial Resistance
- Bacteriocin Research
Background:
- Antibiotic-resistant bacteria and biofilms pose significant clinical challenges.
- Standardized susceptibility testing for sessile bacteria is lacking.
- Bacteriocins show potential as novel antimicrobial agents but require further study against biofilms.
Purpose of the Study:
- To develop a high-throughput, affordable, and simple method for biofilm formation on a 3D surface.
- To evaluate the antimicrobial susceptibility of biofilms using this new method.
- To assess the efficacy of bacteriocins (nisin and enterocin AS-48) against Gram-positive bacterial biofilms.
Main Methods:
- Developed a biofilm growth methodology using glass beads in 96-well microtiter plates.
- Tested biofilm formation and susceptibility with Gram-positive and Gram-negative bacteria.
- Benchmarked the method against the Calgary Biofilm Device.
- Purified and tested bacteriocins nisin and enterocin AS-48 against biofilms.
Main Results:
- Optimized biofilm formation occurred after 48-72 hours incubation.
- The glass bead method demonstrated comparable adhesion and susceptibility to the Calgary Biofilm Device.
- Bacteriocins nisin and AS-48 showed increased efficacy against biofilms compared to planktonic cells.
- Specific eradication concentrations were determined for nisin and AS-48 against Enterococcus faecalis and Staphylococcus aureus.
Conclusions:
- The glass bead method offers a standardized and efficient approach for biofilm susceptibility testing.
- Bacteriocins nisin and AS-48 exhibit significant potential for combating antibiotic-resistant Gram-positive bacterial biofilms.
- Further therapeutic development of bacteriocins is warranted based on their biofilm eradication capabilities.
