An in vitro biofilm model to examine the effect of antibiotic ointments on biofilms produced by burn wound bacterial isolates

• Published in: Burns 2011-03, Vol.37 (2), p.312-321
• Main Authors: Hammond, Adrienne A, Miller, Kyle G, Kruczek, Cassandra J, Dertien, Janet, Colmer-Hamood, Jane A, Griswold, John A, Horswill, Alexander R, Hamood, Abdul N
• Format: Article
• Language: English
• Subjects: Agar; Analysis of Variance; Anti-Bacterial Agents - pharmacology; Antibiotics; Antimicrobial agents; Bacitracin - therapeutic use; Biofilm; Biofilms; Biofilms - drug effects; Biofilms - growth & development; Blood; Burn infection; Burns; Burns - microbiology; Cellulose; Clinical isolates; Critical Care; Drug Therapy, Combination; Gentamicin; Gentamicins - pharmacology; Humans; Microbial Sensitivity Tests - methods; Microorganisms; Mupirocin; Mupirocin - pharmacology; Neomycin - therapeutic use; Ointments; Ointments - pharmacology; Pathogens; Polymyxin B - therapeutic use; Pseudomonas aeruginosa; Pseudomonas aeruginosa - drug effects; Pseudomonas aeruginosa - physiology; Staphylococcus aureus; Staphylococcus aureus - drug effects; Staphylococcus aureus - physiology; Topical antimicrobials; Wounds
• ISSN: 0305-4179; 1879-1409
• DOI: 10.1016/j.burns.2010.09.017

Abstract Purpose Topical treatment of burn wounds is essential as reduced blood supply in the burned tissues restricts the effect of systemic antibiotics. On the burn surface, microorganisms exist within a complex structure termed a biofilm, which enhances bacterial resistance to antimicrobial agents significantly. Since bacteria differ in their ability to develop biofilms, the susceptibility of these biofilms to topically applied antibiotics varies, making it essential to identify which topical antibiotics efficiently disrupt or prevent biofilms produced by these pathogens. Yet, a simple in vitro assay to compare the susceptibility of biofilms produced by burn wound isolates to different topical antibiotics has not been reported. Methods Biofilms were developed by inoculating cellulose disks on agar plates with burn wound isolates and incubating for 24 h. The biofilms were then covered for 24 h with untreated gauze or gauze coated with antibiotic ointment and remaining microorganisms were quantified and visualized microscopically. Results Mupirocin and triple antibiotic ointments significantly reduced biofilms produced by the Staphylococcus aureus and Pseudomonas aeruginosa burn wound isolates tested, as did gentamicin ointment, with the exception of one P. aeruginosa clinical isolate. Conclusions The described assay is a practical and reproducible approach to identify topical antibiotics most effective in eliminating biofilms produced by burn wound isolates.