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Background <p><i>Pseudomonas aeruginosa</i> is an opportunistic pathogen of clinical significance in both human and veterinary medicine. In human healthcare, it is primarily associated with multidrug-resistant hospital-acquired infections. In companion animals, it commonly causes chronic and difficult-to-treat infections of the ears, wounds, and respiratory tract.</p> <p>This study aimed to evaluate the impact of sub-inhibitory concentrations of commonly used antiseptics in veterinary practice on the biofilm-forming ability of <i>P. aeruginosa</i> strains of animal origin.</p> Methods <p>Ten <i>P. aeruginosa</i> strains of animal origin (dogs, cats) isolated from clinical samples as a primary cause of infection, were selected for analysis. Antibiotic susceptibility of the tested strains was determined using the disk diffusion method in accordance with EUCAST and CLSI guidelines. The minimum inhibitory concentrations for two antiseptics: Octenisept and Polisept Vet Wound Irrigation Silver Pro were determined using the microdilution method. Subsequently, the effects of 1/4 and 1/8 MIC concentrations on biofilm formation and its early structure were assessed. Additionally, we evaluated the impact of a short-term exposure to MIC concentrations of tested agents on mature <i>P. aeruginosa</i> biofilms. Fluorescence microscopy was employed to visualize differences in biofilm architecture after 48&#xa0;h of exposure to the antiseptics in 1/4 MIC concentrations.</p> Results <p>All analyzed strains were resistant to amoxicillin–clavulanic acid, tetracycline, and trimethoprim/sulfamethoxazole. The minimum inhibitory concentration values were 0.00625% for Octenisept (Schülke &amp; Mayr, Norderstedt, Germany) and 1.25% for Polisept Vet Wound Irrigation Silver Pro (JM Sante Pharma, Sosnowiec, Poland). Exposure to sub-inhibitory concentrations (1/4 and 1/8 MIC) significantly affected biofilm formation in a time-dependent manner. A marked stimulation of biofilm production was observed after 48&#xa0;h of incubation, whereas this effect was less pronounced after 24&#xa0;h. In contrast, the analyzed antiseptics did not affect the structure of preformed biofilm after 24&#xa0;h of exposure. Fluorescence analysis demonstrated similar proportions of live and dead cells between groups; however, the total number of cells differed substantially, with more than a twofold increase observed under the tested conditions.</p> Conclusions <p>Sub-inhibitory concentrations of antiseptics were found to stimulate biofilm formation in <i>P. aeruginosa</i> strains. Given the potential for low concentrations of antiseptics to occur in the environment, further research and close monitoring of this phenomenon are necessary.</p>

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Preliminary study on the effects of sub-MIC concentrations of octenidine and polyhexanidine on biofilms produced by animal isolates of Pseudomonas aeruginosa

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  • 发表日期：

    2026年3月26日

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  • Paweł Foksiński,
  • Edyta Kaczorek-Łukowska,
  • Natalia Szyryńska,
  • Joanna Małaczewska

摘要

Background

Pseudomonas aeruginosa is an opportunistic pathogen of clinical significance in both human and veterinary medicine. In human healthcare, it is primarily associated with multidrug-resistant hospital-acquired infections. In companion animals, it commonly causes chronic and difficult-to-treat infections of the ears, wounds, and respiratory tract.

This study aimed to evaluate the impact of sub-inhibitory concentrations of commonly used antiseptics in veterinary practice on the biofilm-forming ability of P. aeruginosa strains of animal origin.

Methods

Ten P. aeruginosa strains of animal origin (dogs, cats) isolated from clinical samples as a primary cause of infection, were selected for analysis. Antibiotic susceptibility of the tested strains was determined using the disk diffusion method in accordance with EUCAST and CLSI guidelines. The minimum inhibitory concentrations for two antiseptics: Octenisept and Polisept Vet Wound Irrigation Silver Pro were determined using the microdilution method. Subsequently, the effects of 1/4 and 1/8 MIC concentrations on biofilm formation and its early structure were assessed. Additionally, we evaluated the impact of a short-term exposure to MIC concentrations of tested agents on mature P. aeruginosa biofilms. Fluorescence microscopy was employed to visualize differences in biofilm architecture after 48 h of exposure to the antiseptics in 1/4 MIC concentrations.

Results

All analyzed strains were resistant to amoxicillin–clavulanic acid, tetracycline, and trimethoprim/sulfamethoxazole. The minimum inhibitory concentration values were 0.00625% for Octenisept (Schülke & Mayr, Norderstedt, Germany) and 1.25% for Polisept Vet Wound Irrigation Silver Pro (JM Sante Pharma, Sosnowiec, Poland). Exposure to sub-inhibitory concentrations (1/4 and 1/8 MIC) significantly affected biofilm formation in a time-dependent manner. A marked stimulation of biofilm production was observed after 48 h of incubation, whereas this effect was less pronounced after 24 h. In contrast, the analyzed antiseptics did not affect the structure of preformed biofilm after 24 h of exposure. Fluorescence analysis demonstrated similar proportions of live and dead cells between groups; however, the total number of cells differed substantially, with more than a twofold increase observed under the tested conditions.

Conclusions

Sub-inhibitory concentrations of antiseptics were found to stimulate biofilm formation in P. aeruginosa strains. Given the potential for low concentrations of antiseptics to occur in the environment, further research and close monitoring of this phenomenon are necessary.