Time-dependent effects of rifampicin on staphylococcal biofilms

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.

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 (ODr = 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.

The incubation of the biofilms with rifampicin led to a significant reduction of the OD of the biofilms within 1 minute (ODr baseline: 1; ODr 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.

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.