Antibiofilm Potential and Mechanisms of Lacticaseibacillus paracasei L475 Against Multidrug-Resistant Escherichia coli Isolated from Older Adults

The high prevalence of biofilm-associated multidrug-resistant (MDR) Escherichia coli infections in older adults calls for novel control strategies. This study compared fecal E. coli carriage, antimicrobial resistance, and biofilm formation among community-dwelling older adults with different self-reported immune statuses (lower vs. normal), and evaluated the antibiofilm activity of five Lactobacillus cell-free supernatants (CFSs). Fecal samples from 20 older adults were analyzed. E. coli was enumerated, and isolates were characterized for antimicrobial susceptibility and biofilm formation. Five Lactobacillus strains were screened for antibiofilm activity using crystal violet assay, with further evaluation of extracellular polymeric substance (EPS) production and biofilm morphology. After removing the redundant isolates, 70 isolates were reported, with significantly higher counts in the lower-immunity group (7.89 vs. 6.04 log MPN/g). The lower-immunity group had significantly higher antimicrobial resistance (97.3% vs. 60.6%), and higher MDR prevalence (91.7% vs. 24.2%). Biofilm formation was observed in 62.9% of isolates, with significantly higher prevalence among MDR isolates and in the lower-immunity group. L. paracasei L475 CFS showed the strongest antibiofilm activity against a representative MDR isolate (L5-1), with inhibition and eradication rates of 82.9% and 75.0%, respectively. Mechanistically, L475 CFS reduced extracellular polymeric substance components, with a 92.3% reduction in proteins and 41.3% in polysaccharides. Microscopy confirmed disrupted biofilm architecture, membrane damage, and cell lysis. In conclusion, these preliminary findings indicate a potential association between self-reported immune function and E. coli resistance/biofilm formation in older adults. L. paracasei L475 CFS demonstrates promising in vitro antibiofilm activity against an MDR E. coli isolate from this population, supporting its potential as a postbiotic candidate.