Time-Dependent Effects of Rifampicin on Staphylococcal Biofilms

Abstract

Objective

To test the time-dependent effects of rifampicin on established biofilms of Staphylococcus epidermidis isolated from patients with cardiac implant infections and catheter-related bacteremia.

Methods

Biofilms were grown in microtiter plates for 24 hours, dyed and stained with crystal violet. The mean optical density (OD) was used for quantification. The OD ratio (OD_r = OD of the treated biofilm/OD of the untreated biofilm) was used to measure changes in the thickness of the biofilms over the time. Biofilms were incubated with rifampicin (0.6 mg/mL) for 1, 5, 15, 30 and 60 minutes. Unstained biofilms were sonicated and plated on Columbia agar for time-kill curves.

Results

The incubation of the biofilms with rifampicin led to a significant reduction of the OD of the biofilms within 1 minute (OD_r baseline: 1; OD_r 1 min: 0.333 ± 0.131) (p<0.001). With regard to bacterial killing, rifampicin reduced the mean log count, but viable bacteria were still grown from biofilms in 13 out of 28 isolates despite MIC values < 0.01 mg/L.

Conclusions

In conclusion, our results confirm that rifampicin at a concentration of 1.2 mg/mL immediately reduces established biofilms formed by S. epidermidis although it is not bactericidal despite very low MICs at planktonic conditions.