1887

Abstract

is predominant in implant-associated infections due to its capability to form biofilms. It can deploy several strategies for biofilm development using either polysaccharide intercellular adhesin (PIA), extracellular DNA (eDNA) and/or proteins, such as the extracellular matrix-binding protein (Embp).

We hypothesize that the dichotomic regulation of adhesins is linked to whether it is inside a host or not, and that biofilm investigations in laboratory media may not reflect actual biofilms .

We address the importance of PIA and Embp in biofilm grown in ‘humanized’ media to understand if these components play different roles in biofilm formation under conditions where bacteria can incorporate host proteins in the biofilm matrix.

1585 WT (deficient in ), and derivative strains that either lack , express from an inducible promotor, or express from a plasmid, were cultivated in standard laboratory media, or in media with human plasma or serum. The amount, structure, elasticity and antimicrobial penetration of biofilms was quantified to describe structural differences caused by the different matrix components and growth conditions. Finally, we quantified the initiation of biofilms as suspended aggregates in response to host factors to determine how quickly the cells aggregate in response to the host environment and reach a size that protects them from phagocytosis.

1585 required polysaccharides to form biofilm in laboratory media. However, these observations were not representative of the biofilm phenotype in the presence of human plasma. If human plasma were present, polysaccharides and Embp were redundant for biofilm formation. Biofilms formed in human plasma were loosely attached and existed mostly as suspended aggregates. Aggregation occurred after 2 h of exposing cells to plasma or serum. Despite stark differences in the amount and composition of biofilms formed by polysaccharide-producing and Embp-producing strains in different media, there were no differences in vancomycin penetration or susceptibility.

We suggest that the assumed importance of polysaccharides for biofilm formation is an artefact from studying biofilms in laboratory media void of human matrix components. The cell–cell aggregation of can be activated by host factors without relying on either of the major adhesins, PIA and Embp, indicating a need to revisit the basic question of how deploys self-produced and host-derived matrix components to form antibiotic-tolerant biofilms .

Funding
This study was supported by the:
  • Sundhedsvidenskabelige Fakultet, Aarhus Universitet (Award PhD grant)
    • Principle Award Recipient: SandraM. Skovdal
  • Carlsbergfondet (DK) (Award CF16-0342)
    • Principle Award Recipient: RikkeLouise Meyer
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License.
Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.001287
2021-01-25
2021-10-16
Loading full text...

Full text loading...

/deliver/fulltext/jmm/70/3/jmm001287.html?itemId=/content/journal/jmm/10.1099/jmm.0.001287&mimeType=html&fmt=ahah

References

  1. Kloos WE, Musselwhite MS. Distribution and persistence of Staphylococcus and Micrococcus species and other aerobic bacteria on human skin. Appl Microbiol 1975; 30:381–395 [View Article][PubMed]
    [Google Scholar]
  2. Grice EA, Kong HH, Conlan S, Deming CB, Davis J et al. Topographical and temporal diversity of the human skin microbiome. Science 2009; 324:1190–1192 [View Article][PubMed]
    [Google Scholar]
  3. Otto M. Staphylococcus epidermidis--the 'accidental' pathogen. Nat Rev Microbiol 2009; 7:555567 [View Article][PubMed]
    [Google Scholar]
  4. Rogers KL, Fey PD, Rupp ME. Coagulase-Negative staphylococcal infections. Infect Dis Clin North Am 2009; 23:73–98 [View Article]
    [Google Scholar]
  5. Otto M. Staphylococcus epidermidis Pathogenesis. In Fey PD. editor Staphylococcus Epidermidis: Methods and Protocols Totowa, NJ: Humana Press; 2014 pp 17–31
    [Google Scholar]
  6. Rupp ME. Clinical characteristics of infections in humans due to Staphylococcus epidermidis . In Fey PD. editor Staphylococcus Epidermidis: Methods and Protocols Totowa, NJ: Humana Press; 2014. pp 1–16
    [Google Scholar]
  7. Rupp ME, Archer GL. Coagulase-negative staphylococci: pathogens associated with medical progress. Clin Infect Dis 1994; 19:231–245 [View Article][PubMed]
    [Google Scholar]
  8. Conlon KM, Humphreys H, O'Gara JP, O’Gara JP. Regulation of icaR gene expression in Staphylococcus epidermidis . FEMS Microbiol Lett 2002; 216:171–177 [View Article][PubMed]
    [Google Scholar]
  9. Cue D, Lei MG, Lee CY. Genetic regulation of the intercellular adhesion locus in staphylococci. Front Cell Infect Microbiol 2012; 2:38 [View Article][PubMed]
    [Google Scholar]
  10. Büttner H, Mack D, Rohde H. Structural basis of Staphylococcus epidermidis biofilm formation: mechanisms and molecular interactions. Front Cell Infect Microbiol 2015; 5:14 [View Article][PubMed]
    [Google Scholar]
  11. Heilmann C, Schweitzer O, Gerke C, Vanittanakom N, Mack D et al. Molecular basis of intercellular adhesion in the biofilm-forming Staphylococcus epidermidis . Mol Microbiol 1996; 20:1083–1091 [View Article][PubMed]
    [Google Scholar]
  12. Mack D, Nedelmann M, Krokotsch A, Schwarzkopf A, Heesemann J et al. Characterization of transposon mutants of biofilm-producing Staphylococcus epidermidis impaired in the accumulative phase of biofilm production: genetic identification of a hexosamine-containing polysaccharide intercellular adhesin. Infect Immun 1994; 62:3244–3253 [View Article][PubMed]
    [Google Scholar]
  13. Conlon KM, Humphreys H, O'Gara JP, O’Gara JP. icaR encodes a transcriptional repressor involved in environmental regulation of ICA operon expression and biofilm formation in Staphylococcus epidermidis . J Bacteriol 2002; 184:4400–4408 [View Article][PubMed]
    [Google Scholar]
  14. Christner M, Franke GC, Schommer NN, Wendt U, Wegert K et al. The giant extracellular matrix-binding protein of Staphylococcus epidermidis mediates biofilm accumulation and attachment to fibronectin. Mol Microbiol 2010; 75:187–207 [View Article][PubMed]
    [Google Scholar]
  15. Decker R, Burdelski C, Zobiak M, Büttner H, Franke G et al. An 18 kDa scaffold protein is critical for Staphylococcus epidermidis biofilm formation. PLoS Pathog 2015; 11:e1004735 [View Article][PubMed]
    [Google Scholar]
  16. Christner M, Heinze C, Busch M, Franke G, Hentschke M et al. sarA negatively regulates Staphylococcus epidermidis biofilm formation by modulating expression of 1 MDa extracellular matrix binding protein and autolysis-dependent release of eDNA. Mol Microbiol 2012; 86:394–410 [View Article][PubMed]
    [Google Scholar]
  17. Schommer NN, Christner M, Hentschke M, Ruckdeschel K, Aepfelbacher M et al. Staphylococcus epidermidis uses distinct mechanisms of biofilm formation to interfere with phagocytosis and activation of mouse macrophage-like cells 774A.1. Infect Immun 2011; 79:2267–2276 [View Article][PubMed]
    [Google Scholar]
  18. Rohde H, Burandt EC, Siemssen N, Frommelt L, Burdelski C et al. Polysaccharide intercellular adhesin or protein factors in biofilm accumulation of Staphylococcus epidermidis and Staphylococcus aureus isolated from prosthetic hip and knee joint infections. Biomaterials 2007; 28:1711–1720 [View Article][PubMed]
    [Google Scholar]
  19. Andrey DO, Jousselin A, Villanueva M, Renzoni A, Monod A et al. Impact of the regulators SigB, rot, SARA and sarS on the toxic shock Tst promoter and TSST-1 expression in Staphylococcus aureus . PLoS One 2015; 10:e0135579 [View Article][PubMed]
    [Google Scholar]
  20. Pintens V, Massonet C, Merckx R, Vandecasteele S, Peetermans WE et al. The role of sigmaB in persistence of Staphylococcus epidermidis foreign body infection. Microbiology 2008; 154:2827–2836 [View Article][PubMed]
    [Google Scholar]
  21. Olwal Charles Ochieng', Ang'ienda Paul Oyieng', Ochiel DO, Olwal CO, Ang’ienda PO. Alternative sigma factor BB) and catalase enzyme contribute to Staphylococcus epidermidis biofilm's tolerance against physico-chemical disinfection. Sci Rep 2019; 9:5355 [View Article][PubMed]
    [Google Scholar]
  22. Ommen P, Zobek N, Meyer RL. Quantification of biofilm biomass by staining: non-toxic safranin can replace the popular crystal violet. J Microbiol Methods 2017; 141:87–89 [View Article][PubMed]
    [Google Scholar]
  23. Zeng G, Vad BS, Dueholm MS, Christiansen G, Nilsson M et al. Functional bacterial amyloid increases Pseudomonas biofilm hydrophobicity and stiffness. Front Microbiol 2015; 6:1099 [View Article][PubMed]
    [Google Scholar]
  24. Schneider CA, Rasband WS, Eliceiri KW. Nih image to ImageJ: 25 years of image analysis. Nat Methods 2012; 9:671–675 [View Article][PubMed]
    [Google Scholar]
  25. Secor PR, Michaels LA, Ratjen A, Jennings LK, Singh PK. Entropically driven aggregation of bacteria by host polymers promotes antibiotic tolerance in Pseudomonas aeruginosa. Proc Natl Acad Sci U S A 2018; 16;115:10780–10785
    [Google Scholar]
  26. Brust M, Aouane O, Thiébaud M, Flormann D, Verdier C et al. The plasma protein fibrinogen stabilizes clusters of red blood cells in microcapillary flows. Sci Rep 2015; 4:4348 [View Article]
    [Google Scholar]
  27. Rügheimer L, Hansell P, Wolgast M. Determination of the charge of the plasma proteins and consequent Donnan equilibrium across the capillary barriers in the rat microvasculature. Acta Physiologica 2008; 194:335–339 [View Article]
    [Google Scholar]
  28. Campoccia D, Montanaro L, Ravaioli S, Pirini V, Cangini I et al. Exopolysaccharide production by Staphylococcus epidermidis and its relationship with biofilm extracellular DNA. Int J Artif Organs 2011; 34:832–839 [View Article]
    [Google Scholar]
  29. Singh R, Sahore S, Kaur P, Rani A, Ray P. Penetration barrier contributes to bacterial biofilm-associated resistance against only select antibiotics, and exhibits genus-, strain- and antibiotic-specific differences. Pathog Dis 2016; 74:ftw056 [View Article]
    [Google Scholar]
  30. Singh R, Ray P, Das A, Sharma M. Penetration of antibiotics through Staphylococcus aureus and Staphylococcus epidermidis biofilms. J Antimicrob Chemother 2010; 65:1955–1958 [View Article]
    [Google Scholar]
  31. Kristian SA, Birkenstock TA, Sauder U, Mack D, Götz F et al. Biofilm formation induces C3a release and protects Staphylococcus epidermidis from IgG and complement deposition and from neutrophil-dependent killing. J Infect Dis 2008; 197:1028–1035 [View Article]
    [Google Scholar]
  32. Champion JA, Mitragotri S. Role of target geometry in phagocytosis. Proc Natl Acad Sci U S A 2006; 103:4930–4934 [View Article]
    [Google Scholar]
  33. Crosby HA, Kwiecinski J, Horswill AR. Staphylococcus aureus aggregation and coagulation mechanisms, and their function in host-pathogen interactions. Adv Appl Microbiol 2016; 96:1–41
    [Google Scholar]
  34. Rohde H, Burdelski C, Bartscht K, Hussain M, Buck F et al. Induction of Staphylococcus epidermidis biofilm formation via proteolytic processing of the accumulation-associated protein by staphylococcal and host proteases. Mol Microbiol 2005; 55:1883–1895 [View Article][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.001287
Loading
/content/journal/jmm/10.1099/jmm.0.001287
Loading

Data & Media loading...

Supplements

Supplementary material 1

PDF

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error