Abstract

Staphylococcus aureus is one of the most relevant opportunistic pathogens involved in many biofilm-associated diseases, and is a major cause of nosocomial infections, mainly due to the increasing prevalence of multidrug-resistant strains. Consequently, alternative methods to eradicate the pathogen are urgent. It has been previously shown that polyvalent staphylococcal kayviruses and their derived endolysins are excellent candidates for therapy. Here we present the characterization of a new bacteriophage: vB_SauM-LM12 (LM12). LM12 has a broad host range (>90%; 56 strains tested), and is active against several MRSA strains. The genome of LM12 is composed of a dsDNA molecule with 143,625 bp, with average GC content of 30.25% and codes for 227 Coding Sequences (CDSs). Bioinformatics analysis did not identify any gene encoding virulence factors, toxins, or antibiotic resistance determinants. Antibiofilm assays have shown that this phage significantly reduced the number of viable cells (less than one order of magnitude). Moreover, the encoded endolysin also showed activity against biofilms, with a consistent biomass reduction during prolonged periods of treatment (of about one order of magnitude). Interestingly, the endolysin was shown to be much more active against stationary-phase cells and suspended biofilm cells than against intact and scraped biofilms, suggesting that cellular aggregates protected by the biofilm matrix reduced protein activity. Both phage LM12 and its endolysin seem to have a strong antimicrobial effect and broad host range against S. aureus, suggesting their potential to treat S. aureus biofilm infections.

Highlights

  • The development of bacterial resistance to antibiotics has become a global concern.Very recently, the World Health Organization (WHO) published a list of priority pathogens resistant to antibiotics, encouraging the scientific community and pharmaceutical industries to focus on the development of new antimicrobials to combat antimicrobial-resistant (AMR) pathogens [1]

  • We report the isolation of a new Kayvirus phage, named vB_SauM-LM12 (LM12)

  • Twenty-six methicillin-resistant Staphylococcus aureus (MRSA) clinical strains were used for phage enrichment, using wastewater treatment plant sewage samples as phage sources

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Summary

Introduction

The World Health Organization (WHO) published a list of priority pathogens resistant to antibiotics, encouraging the scientific community and pharmaceutical industries to focus on the development of new antimicrobials to combat antimicrobial-resistant (AMR) pathogens [1]. Antibiotic-resistant S. aureus is well-established in both community and healthcare settings, being one the most frequently isolated pathogens in hospital-associated infections (HAI) [2]. This microorganism is a resourceful pathogen that can cause a wide range of diseases, from food poisoning to life-threatening diseases such as pneumonia, infective endocarditis, or sepsis [3]. The severity of S. aureus infections is mostly related to its intrinsic ability to acquire and express antibiotic-resistance genes.

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