Inhibitory Effect of Antimicrobial Peptides Bac7(17), PAsmr5-17 and PAβN on Bacterial Growth and Biofilm Formation of Multidrug-Resistant Acinetobacter baumannii.

Abstract

Acinetobacter (A.) baumannii is a major nosocomial pathogen in human and veterinary medicine. The emergence of certain international clones (ICs), often with multidrug-resistant (MDR) phenotypes and biofilm formation (BF), facilitates its spread in clinical environments. The global rise in antimicrobial resistance demands alternative treatment strategies, such as antimicrobial peptides (AMPs). In this study, 45 human and companion animal MDR-A. baumannii isolates, belonging to the globally spread IC1, IC2 and IC7, were tested for antimicrobial resistance and biofilm-associated genes (BAGs) and their capacity for BF. Of these, 13 were used to test the inhibitory effect of AMPs on bacterial growth (BG) and BF through the application of a crystal violet assay. The two novel AMP variants Bac7(17) (target cell inactivation) and Pasmr5-17 (efflux pump inhibition) and the well-known AMP phenylalanine-arginine-β-naphthylamide (PAβN) were tested at concentrations of 1.95 to 1000 µg/mL. Based on whole-genome sequence data, identical patterns of BAGs were detected within the same IC. AMPs inhibited BG and BF in a dose-dependent manner. Bac7(17) and PAsmr5-17 were highly effective against BG, with growth inhibition (GI) of >99% (62.5 and 125 µg/mL, respectively). PAβN achieved only 95.7% GI at 1000 µg/mL. Similar results were obtained for BF. Differences between the ICs were found for both GI and BF when influenced by AMPs. PAsmr5-17 had hardly any inhibitory effect on the BF of IC1 isolates, but for IC2 and IC7 isolates, 31.25 µg/mL was sufficient. Our data show that the susceptibility of animal MDR-A. baumannii to AMPs most likely resembles that of human isolates, depending on their assignment to a particular IC. Even low concentrations of AMPs had a significant effect on BG. Therefore, AMPs represent a promising alternative in the treatment of MDR-A. baumannii, either as the sole therapy or in combination with antibiotics.