Effect of Cinnamon Oil on icaA Expression and Biofilm Formation by Staphylococcus epidermidis

• Published in: Applied and Environmental Microbiology 2009-11, Vol.75 (21), p.6850-6855
• Main Authors: Nuryastuti, Titik, van der Mei, Henny C, Busscher, Henk J, Iravati, Susi, Aman, Abu T, Krom, Bastiaan P
• Format: Article
• Language: English
• Subjects: agar; Anti-Bacterial Agents - pharmacology; antibacterial properties; Bacteria; Bacterial Proteins - biosynthesis; biofilm; Biofilms; Biofilms - drug effects; bromides; Cell culture; chlorhexidine; cinnamon; Drug Synergism; Effects; Environmental Microbiology; essential oils; gene expression; Gene Expression Profiling; Gene Expression Regulation, Bacterial - drug effects; gentamicin; Humans; image analysis; medical equipment; microbial growth; Microbial Sensitivity Tests; Microbial Viability; microscopy; minimum inhibitory concentration; Oils & fats; Oils, Volatile - pharmacology; polymerase chain reaction; Staphylococcal Infections; Staphylococcus epidermidis; Staphylococcus epidermidis - drug effects; Staphylococcus epidermidis - isolation & purification; Staphylococcus epidermidis - physiology; Streptococcus infections; tetrazolium; Tetrazolium Salts - metabolism; Thiazoles - metabolism
• ISSN: 0099-2240; 1098-5336; 1098-6596
• DOI: 10.1128/AEM.00875-09

Staphylococcus epidermidis is notorious for its biofilm formation on medical devices, and novel approaches to prevent and kill S. epidermidis biofilms are desired. In this study, the effect of cinnamon oil on planktonic and biofilm cultures of clinical S. epidermidis isolates was evaluated. Initially, susceptibility to cinnamon oil in planktonic cultures was compared to the commonly used antimicrobial agents chlorhexidine, triclosan, and gentamicin. The MIC of cinnamon oil, defined as the lowest concentration able to inhibit visible microbial growth, and the minimal bactericidal concentration, the lowest concentration required to kill 99.9% of the bacteria, were determined using the broth microdilution method and plating on agar. A checkerboard assay was used to evaluate the possible synergy between cinnamon oil and the other antimicrobial agents. The effect of cinnamon oil on biofilm growth was studied in 96-well plates and with confocal laser-scanning microscopy (CLSM). Biofilm susceptibility was determined using a metabolic 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Real-time PCR analysis was performed to determine the effect of sub-MIC concentrations of cinnamon oil on expression of the biofilm-related gene, icaA. Cinnamon oil showed antimicrobial activity against both planktonic and biofilm cultures of clinical S. epidermidis strains. There was only a small difference between planktonic and biofilm MICs, ranging from 0.5 to 1% and 1 to 2%, respectively. CLSM images indicated that cinnamon oil is able to detach and kill existing biofilms. Thus, cinnamon oil is an effective antimicrobial agent to combat S. epidermidis biofilms.