title: Metabolic activity, urease production, antibiotic resistance and virulence in dual species biofilms of Staphylococcus epidermidis and Staphylococcus aureus.
abstract: In this paper, the metabolic activity in single and dual species biofilms of Staphylococcus epidermidis and Staphylococcus aureus isolates was investigated. Our results demonstrated that there was less metabolic activity in dual species biofilms compared to S. aureus biofilms. However, this was not observed if S. aureus and S. epidermidis were obtained from the same sample. The largest effect on metabolic activity was observed in biofilms of S. aureus Mu50 and S. epidermidis ET-024. A transcriptomic analysis of these dual species biofilms showed that urease genes and genes encoding proteins involved in metabolism were downregulated in comparison to monospecies biofilms. These results were subsequently confirmed by phenotypic assays. As metabolic activity is related to acid production, the pH in dual species biofilms was slightly higher compared to S. aureus Mu50 biofilms. Our results showed that S. epidermidis ET-024 in dual species biofilms inhibits metabolic activity of S. aureus Mu50, leading to less acid production. As a consequence, less urease activity is required to compensate for low pH. Importantly, this effect was biofilm-specific. Also S. aureus Mu50 genes encoding virulence-associated proteins (Spa, SplF and Dps) were upregulated in dual species biofilms compared to monospecies biofilms and using Caenorhabditis elegans infection assays, we demonstrated that more nematodes survived when co-infected with S. epidermidis ET-024 and S. aureus mutants lacking functional spa, splF or dps genes, compared to nematodes infected with S. epidermidis ET-024 and wild- type S. aureus. Finally, S. epidermidis ET-024 genes encoding resistance to oxacillin, erythromycin and tobramycin were upregulated in dual species biofilms and increased resistance was subsequently confirmed. Our data indicate that both species in dual species biofilms of S. epidermidis and S. aureus influence each other's behavior, but additional studies are required necessary to elucidate the exact mechanism(s) involved.
authors.affiliation: Laboratorium voor Farmaceutische Microbiologie, Universiteit Gent, Ottergemsesteenweg, Gent, Belgium.
authors.forename: Ilse
authors.initials: I
authors.lastname: Vandecandelaere
authors.affiliation: Laboratorium voor Farmaceutische Biotechnologie, Universiteit Gent, Ottergemsesteenweg, Gent, Belgium.
authors.forename: Filip
authors.initials: F
authors.lastname: Van Nieuwerburgh
authors.affiliation: Laboratorium voor Farmaceutische Biotechnologie, Universiteit Gent, Ottergemsesteenweg, Gent, Belgium.
authors.forename: Dieter
authors.initials: D
authors.lastname: Deforce
authors.affiliation: Laboratorium voor Farmaceutische Microbiologie, Universiteit Gent, Ottergemsesteenweg, Gent, Belgium.
authors.forename: Tom
authors.identifier: http://orcid.org/0000-0002-6407-0601
authors.initials: T
authors.lastname: Coenye
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D018441:Biofilms / Q000187:drug effects*; D017173:Caenorhabditis elegans / Q000382:microbiology; D024881:Drug Resistance, Bacterial; D004734:Energy Metabolism; D004789:Enzyme Activation; D020869:Gene Expression Profiling; D006801:Humans; D006863:Hydrogen-Ion Concentration; D013203:Staphylococcal Infections / Q000382:microbiology; D013211:Staphylococcus aureus / Q000187:drug effects* / Q000302:isolation & purification / Q000502:physiology*; D013212:Staphylococcus epidermidis / Q000187:drug effects* / Q000302:isolation & purification / Q000502:physiology*; D059467:Transcriptome; D014510:Urease / Q000096:biosynthesis*; D014774:Virulence / Q000235:genetics
chemical_list: D000900:Anti-Bacterial Agents; D014510:Urease