Abstract

Staphylococcal biofilms are a major concern in both clinical and food settings because they are an important source of contamination. The efficacy of established cleaning procedures is often hindered due to the ability of some antimicrobial compounds to induce biofilm formation, and to the presence of persister cells, a small bacterial subpopulation that exhibits multidrug tolerance. Phage lytic enzymes have demonstrated antimicrobial activity against planktonic and sessile bacteria. However, their ability to lyse and/or select persister cells remains largely unexplored so far. In this work, the lytic activity of the endolysin LysH5 against Staphylococcus aureus and Staphylococcus epidermidis biofilms was confirmed. LysH5 reduced staphylococcal sessile cell counts by 1–3 log units, compared with the untreated control, and sub-inhibitory concentrations of this protein did not induce biofilm formation. LysH5-surviving cells were not resistant to the lytic activity of this protein, suggesting that no persister cells were selected. Moreover, to prove the lytic ability of LysH5 against this subpopulation, both S. aureus exponential cultures and persister cells obtained after treatment with rifampicin and ciprofloxacin were subsequently treated with LysH5. The results demonstrated that besides the notable activity of endolysin LysH5 against staphylococcal biofilms, persister cells were also inhibited, which raises new opportunities as an adjuvant for some antibiotics.

Highlights

  • Two staphylococcal species, Staphylococcus aureus and Staphylococcus epidermidis, are the main cause of hospital-associated infections [1,2]

  • LysH5 is effective against staphylococcal biofilms The ability of LysH5 to remove staphylococcal biofilms and kill sessile cells was determined on six S. aureus and three S. epidermidis strains

  • Two S. aureus strains isolated from a food environment and three S. epidermidis of clinical origin, all of them carrying biofilm related genes, were included in the assay (Table 1). 24 h-old biofilms developed on 96well polystyrene plates were treated with LysH5 (0.15 mM) and lysostaphin (0.2 mM) and biofilm removal was determined after 6 h of incubation

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Summary

Introduction

Staphylococcus aureus and Staphylococcus epidermidis, are the main cause of hospital-associated infections [1,2]. S. aureus is one of the major bacterial agents causing foodborne diseases in humans due to the production of enterotoxins [5]. Pathogenicity of both species is clearly associated with their ability to form biofilms on biotic and abiotic surfaces, providing high resistance to host defenses and antibiotics, and to cleaning and disinfection processes [2]. The ability to form biofilms in both species is due to the production of an extracellular material that contributes to intercellular aggregation and attachment to surfaces In most cases, this matrix is composed of exopolysaccharides such as polyN-acetyl-b-(1,6)-glucosamine (PIA/PNAG), teichoic acids, DNA and specific proteins [2]. Specific proteins such as Bap [7], Spa [8], FnBPA, FnBPB [9], and

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