Antimicrobial and Antibiofilm Activities of Citral Against Carbapenem-Resistant Enterobacter cloacae

Citral that is produced in the essential oils of plants is an isomeric mixture of geranial and neral. Few recent studies exhibited robust antibacterial activity of citral against several pathogens. However, little is reported about effects of citral on carbapenem-resistant Enterobacter cloacae (CREC). The purpose of this study was to assess antibacterial and antibiofilm activities of citral against CREC. Here, minimum inhibitory concentrations (MICs) of citral against CREC were determined by the agar dilution method. Antibacterial mode of citral against CREC was elucidated by evaluating changes in intracellular adenosine triphosphate (ATP) concentration, intracellular pH (pHin), membrane potential, membrane integrity, and cell morphology. In addition, CREC cell damage within biofilms was examined by confocal laser scanning microscopy (CLSM). The results showed that the MIC of citral against CREC was 1000 μg/mL. Citral inhibited CREC growth and destroyed its cell membrane integrity, as measured by the decrease of intracellular ATP, pH, and membrane potential as well as distinctive deformation in cellular morphology. CLSM images demonstrated that citral could inactivate CREC cells within biofilms. These results revealed that citral exhibits potent antibacterial and antibiofilm activity against CREC, and thus has potential to be used as natural food preservatives to control CREC-associated infection spread.