Inhibitory effect of trisodium citrate on biofilms formed by Klebsiella pneumoniae.

OBJECTIVES

Klebsiella pneumoniae is a significant nosocomial pathogen related to ventilator-associated pneumonia owing to biofilm formation. Trisodium citrate (TSC) has antibacterial activity, but there is little research on the effect of TSC on biofilm formed by K. pneumoniae. The aims of this study were to evaluate the inhibitory effect of 4% TSC on K. pneumoniae biofilm formation and to determine the best time of TSC addition for biofilm inhibition.

METHODS

A total of 45 K. pneumoniae strains isolated from tracheal tip and sputum specimens were included. Modified Congo red agar was used to screen for biofilm production. Biofilm-positive strains were cultured for 4 days. TSC (4%) was added either initially or 3 days later. Crystal violet staining was used to quantify biofilm mass by measuring the optical density at 570 nm (OD). Scanning electron microscopy (SEM) was used to observe biofilm morphology.

RESULTS

The OD was significantly lower in the 4% TSC group than that in the no-TSC group during the 4-day experiment. Compared with addition of TSC after 3 days, initial TSC addition resulted in a significant absorbance decrease (Day 4, 0.63 ± 0.11 later-TSC group vs. 0.41 ± 0.16 initial-TSC group). As observed by SEM, bacteria were stacked most densely in the no-TSC group on Day 4. In contrast, few bacteria were observed when TSC was added initially, whilst bacteria were obviously dispersed when TSC was added after 3 days.

CONCLUSION

TSC can inhibit K. pneumoniae biofilm formation and has the best effect when added initially.