Constitutive Phenotypic Modification of Lipid A in Clinical Acinetobacter baumannii Isolates.

The degree of polymyxin B (PMB) resistance was measured in 40 clinical Acinetobacter baumannii isolates obtained from health care facilities. All of the tested isolates possessed a multidrug-resistant (MDR) phenotype against four classes of antibiotics (meropenem, doxycycline, gentamicin, and erythromycin), except for PMB. The bla OXA-23 gene was detected throughout the genetic analysis and experimental assay, indicating that all of the MDR strains were carbapenem-resistant A. baumannii strains. Multilocus sequence typing-based genotyping revealed that nine selected strains belonged to the international clone II lineage. When matrix-assisted laser desorption ionization-time of flight mass spectrometry was performed, intrinsic lipid A modification by phosphoethanolamine (PEtN) incorporation was noticeable only in the PMB-resistant (PMB R ) strains. However, the presence of hexa- and penta-acylated lipid A due to the loss of the laurate (C 12 ) acyl chain was noted in all PMB-susceptible strains but not in the PMB R strains. The reduction of negative surface charges in the PMB R strains was assessed by zeta potential analysis. Fluorescence imaging using dansyl-PMB revealed that, in the PMB R strains, PMB was less likely to bind to the cell surface. IMPORTANCE The widespread presence of MDR pathogens, including A. baumannii, is causing serious hospital-acquired infections worldwide. Extensive surveillance of MDR clinical A. baumannii isolates has been conducted, but the underlying mechanisms for their development of MDR phenotypes are often neglected. Either lipid A modification or loss of lipopolysaccharide in Gram-negative bacteria leads to PMB R phenotypes. The prevalence of intrinsic lipid A modification in PMB R clinical strains was attributed to high levels of basal expression of pmrC and eptA-1 . Our findings suggest that new therapeutic strategies are warranted to combat MDR pathogens due to the emergence of many PMB R clinical strains.