Abstract

Host defense peptides or antimicrobial peptides (AMPs), e.g., cathelicidins, have recently been discussed as a potential new treatment option against bacterial infections. To test the efficacy of AMPs, standardized methods that closely mimic the physiological conditions at the site of infection are still needed. The aim of our study was to test the meningitis-causing bacteria Streptococcus suis and Escherichia coli for their susceptibility to cathelicidins in culture medium versus cerebrospinal fluid (CSF). Susceptibility testing was performed in analogy to the broth microdilution method described by the Clinical and Laboratory Standard Institute (CLSI) to determine minimum inhibitory concentrations (MICs) of antimicrobial agents. MICs were determined using cation-adjusted Mueller–Hinton broth (CA-MHB), lysogeny broth (LB), Roswell Park Memorial Institute medium (RPMI) or Dulbecco’s Modified Eagle’s Medium (DMEM) (the latter two supplemented with 5% CA-MHB or blood) and compared with MICs obtained in porcine or human CSF. Our data showed that MICs obtained in CA-MHB as recommended by CLSI do not reflect the MICs obtained in the physiological body fluid CSF. However, the MICs of clinical isolates of S. suis tested in RPMI medium supplemented with CA-MHB, were similar to those of the same strains tested in CSF. In contrast, the MICs in the human CSF for the tested E. coli K1 strain were higher compared to the RPMI medium and showed even higher values than in CA-MHB. This highlights the need for susceptibility testing of AMPs in a medium that closely mimics the clinically relevant conditions.

Highlights

  • The increase in antimicrobial resistance is an emerging problem and requires new strategies to combat bacterial infections (Kim, 2013)

  • The present study focuses on two different meningitis-causing pathogens, the Gram-positive Streptococcus suis and the Gramnegative Escherichia coli

  • The aim of our study was to test the minimum inhibitory concentrations (MICs) of LL-37 and PR-39 with broth microdilution in accordance with the Clinical and Laboratory Standard Institute (CLSI) recommendations (CLSI, 2018) in cation-adjusted Mueller–Hinton broth (CA-Mueller-Hinton broth (MHB)) and compared with lysogeny broth (LB), various well-known cell culture media and human or porcine cerebrospinal fluid (CSF)

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Summary

Introduction

The increase in antimicrobial resistance is an emerging problem and requires new strategies to combat bacterial infections (Kim, 2013). Fighting infections with the application of antimicrobial peptides (AMPs) is considered a promising new strategy. An important group of AMPs are the cathelicidins. They consist of 12–100 amino acids and occur in all vertebrate species (reviewed by Tomasinsig and Zanetti, 2005). Their positive charge allows them to bind to the negatively charged bacterial membranes and subsequently lyse them. Current research is focusing on the synthetic generation of smaller peptidic analogs of cathelicidins with improved antimicrobial activities as useful tools for evaluating cathelicidins in the treatment of severe infections (reviewed by Wang et al, 2019)

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