1887

Abstract

Purpose. The purpose of this study was to develop an alternative, more clinically relevant approach to susceptibility reporting for implant-associated infections. Using 20 staphylococcal isolates, isolated from clinical implant infections, the majority (85 %) demonstrated biofilm-forming capabilities. A significantly increased minimum biofilm eradication concentration (MBEC) compared to minimum inhibitory concentration (MIC) breakpoint was obtained, with MBEC values greater than 256 µg ml for the majority of bacteria. Such a vast increase was also demonstrated for isolates defined as negligible biofilm formers via crystal violet staining, likely due to the high protein content of biofilms, as confirmed by proteinase-K treatment.

Methodology. This study employed a variety of techniques to assess MIC and MBEC of the isolates tested. In addition, the nature of bacterial biofilm across a range of clinical isolates was investigated using crystal violet staining, sodium metaperiodate and proteinase-K treatment, and PCR analysis.

Results/Key findings. Infection of medical implants is associated with increased rates of infection and increased bacterial tolerance to antibiotic strategies. Clinical significance is due to the presence of pathogens attached to biomaterial surfaces enclosed in an extracellular polymeric matrix termed the biofilm. This article highlights the importance of defining the clinical susceptibility of implant-associated infections in vitro using methods that are relevant to the biofilm phenotype in vivo, and highlights how current planktonic-based antimicrobial susceptibility tests are often misleading.

Conclusion. The use of biofilm-relevant susceptibility tests would improve patient outcomes by enabling correct antimicrobial regimens to be rapidly identified, reducing treatment failure and halting the spread of antimicrobial-resistant strains.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.000466
2017-04-28
2019-08-17
Loading full text...

Full text loading...

/deliver/fulltext/jmm/66/4/461.html?itemId=/content/journal/jmm/10.1099/jmm.0.000466&mimeType=html&fmt=ahah

References

  1. Laverty G, Gorman SP, Gilmore BF. Biofilms and implant-associated infections. In Cooper IB, Barnes L. (editors) Biofilms and Implant-Associated Infections, 1st ed. Woodhead Publishing; 2014
    [Google Scholar]
  2. Maki DG, Kluger DM, Crnich CJ. The risk of bloodstream infection in adults with different intravascular devices: a systematic review of 200 published prospective studies. Mayo Clin Proc 2006; 81: 1159– 1171 [CrossRef] [PubMed]
    [Google Scholar]
  3. O'Neill J. Tackling drug resistant infections globally: final report and recommendation. The Review on Antimicrobial Resistance. UK Government and Wellcome Trust; 2016
  4. Andrews JM. BSAC working party on susceptibility testing. BSAC standardized disc susceptibility testing method (version 6). J Antimicrob Chemother 2007; 60: 20– 41 [Crossref]
    [Google Scholar]
  5. Scott RD. 2009; The direct medical costs of healthcare-associated infections in US hospitals and the benefits of prevention. Centres for Disease Control and Prevention www.cdc.gov/HAI/pdfs/hai/Scott_CostPaper.pdf
  6. MacGowan AP, Wise R. Establishing MIC breakpoints and the interpretation of in vitro susceptibility tests. J Antimicrob Chemother 2001; 48: 17– 28 [CrossRef] [PubMed]
    [Google Scholar]
  7. Holá V, Růzicka F, Votava M. [Differences in antibiotic sensitivity in biofilm-positive and biofilm-negative strains of Staphylococcus epidermidis isolated from blood cultures]. Epidemiol Mikrobiol Imunol 2004; 53: 66– 69 [PubMed]
    [Google Scholar]
  8. Widmer AF, Frei R, Rajacic Z, Zimmerli W. Correlation between in vivo and in vitro efficacy of antimicrobial agents against foreign body infections. J Infect Dis 1990; 162: 96– 102 [CrossRef] [PubMed]
    [Google Scholar]
  9. Cerca N, Martins S, Cerca F, Jefferson KK, Pier GB et al. Comparative assessment of antibiotic susceptibility of coagulase-negative staphylococci in biofilm versus planktonic culture as assessed by bacterial enumeration or rapid XTT colorimetry. J Antimicrob Chemother 2005; 56: 331– 336 [CrossRef] [PubMed]
    [Google Scholar]
  10. Dunne WM Jr. Effects of subinhibitory concentrations of vancomycin or cefamandole on biofilm production by coagulase-negative staphylococci. Antimicrob Agents Chemother 1990; 34: 390– 393 [CrossRef] [PubMed]
    [Google Scholar]
  11. Rachid S, Ohlsen K, Witte W, Hacker J, Ziebuhr W. Effect of subinhibitory antibiotic concentrations on polysaccharide intercellular adhesin expression in biofilm-forming Staphylococcus epidermidis. Antimicrob Agents Chemother 2000; 44: 3357– 3363 [CrossRef] [PubMed]
    [Google Scholar]
  12. Sandoe JA, Wysome J, West AP, Heritage J, Wilcox MH. Measurement of ampicillin, vancomycin, linezolid and gentamicin activity against enterococcal biofilms. J Antimicrob Chemother 2006; 57: 767– 770 [CrossRef] [PubMed]
    [Google Scholar]
  13. Stepanovic S, Vukovic D, Dakic I, Savic B, Svabic-Vlahovic M. A modified microtiter-plate test for quantification of staphylococcal biofilm formation. J Microbiol Methods 2000; 40: 175– 179 [CrossRef] [PubMed]
    [Google Scholar]
  14. Mack D, Haeder M, Siemssen N, Laufs R. Association of biofilm production of coagulase-negative staphylococci with expression of a specific polysaccharide intercellular adhesin. J Infect Dis 1996; 174: 881– 883 [CrossRef] [PubMed]
    [Google Scholar]
  15. Wang X, Preston JF 3rd, Romeo T. The pgaABCD locus of Escherichia coli promotes the synthesis of a polysaccharide adhesin required for biofilm formation. J Bacteriol 2004; 186: 2724– 2734 [CrossRef] [PubMed]
    [Google Scholar]
  16. Ziebuhr W, Heilmann C, Götz F, Meyer P, Wilms K et al. Detection of the intercellular adhesion gene cluster (ica) and phase variation in Staphylococcus epidermidis blood culture strains and mucosal isolates. Infect Immun 1997; 65: 890– 896 [PubMed]
    [Google Scholar]
  17. Gorman SP, Adair CG, Mawhinney WM. Incidence and nature of peritoneal catheter biofilm determined by electron and confocal laser scanning microscopy. Epidemiol Infect 1994; 112: 551– 559 [CrossRef] [PubMed]
    [Google Scholar]
  18. Clinical and Laboratory Standards Institute (editor) Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobically; Approved Standard, 10th ed. Clinical and Laboratory Standards Institute; 2015
    [Google Scholar]
  19. Mack D, Becker P, Chatterjee I, Dobinsky S, Knobloch JK et al. Mechanisms of biofilm formation in Staphylococcus epidermidis and Staphylococcus aureus: functional molecules, regulatory circuits, and adaptive responses. Int J Med Microbiol 2004; 294: 203– 212 [CrossRef] [PubMed]
    [Google Scholar]
  20. 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 [CrossRef] [PubMed]
    [Google Scholar]
  21. von Eiff C, Peters G, Heilmann C. Pathogenesis of infections due to coagulase-negative staphylococci. Lancet Infect Dis 2002; 2: 677– 685 [CrossRef] [PubMed]
    [Google Scholar]
  22. Bordi C, de Bentzmann S. Hacking into bacterial biofilms: a new therapeutic challenge. Ann Intensive Care 2011; 1: 19 [CrossRef] [PubMed]
    [Google Scholar]
  23. Cunningham CD 3rd, Slattery WH 3rd, Luxford WM. Postoperative infection in cochlear implant patients. Otolaryngol Head Neck Surg 2004; 131: 109– 114 [CrossRef] [PubMed]
    [Google Scholar]
  24. Isiklar ZU, Darouiche RO, Landon GC, Beck T. Efficacy of antibiotics alone for orthopaedic device related infections. Clin Orthop Relat Res 1996; 332: 184– 189 [CrossRef] [PubMed]
    [Google Scholar]
  25. Raad I, Darouiche R, Hachem R, Sacilowski M, Bodey GP. Antibiotics and prevention of microbial colonization of catheters. Antimicrob Agents Chemother 1995; 39: 2397– 2400 [CrossRef] [PubMed]
    [Google Scholar]
  26. Soroush S, Jabalameli F, Taherikalani M, Amirmozafari N, Fooladi AA et al. Investigation of biofilm formation ability, antimicrobial resistance and the staphylococcal cassette chromosome mec patterns of methicillin resistant Staphylococcus epidermidis with different sequence types isolated from children. Microb Pathog 2016; 93: 126– 130 [CrossRef] [PubMed]
    [Google Scholar]
  27. Blahova J, Kralikova K, Krcmery VS, Babalova M, Menkyna R et al. Four years of monitoring antibiotic resistance in microorganisms from bacteremic patients. J Chemother 2007; 19: 665– 669 [CrossRef] [PubMed]
    [Google Scholar]
  28. Gatermann SG, Koschinski T, Friedrich S. Distribution and expression of macrolide resistance genes in coagulase-negative staphylococci. Clin Microbiol Infect 2007; 13: 777– 781 [CrossRef] [PubMed]
    [Google Scholar]
  29. Frank KL, Reichert EJ, Piper KE, Patel R. In vitro effects of antimicrobial agents on planktonic and biofilm forms of Staphylococcus lugdunensis clinical isolates. Antimicrob Agents Chemother 2007; 51: 888– 895 [CrossRef] [PubMed]
    [Google Scholar]
  30. Patel JD, Ebert M, Ward R, Anderson JM. S. epidermidis biofilm formation: effects of biomaterial surface chemistry and serum proteins. J Biomed Mater Res A 2007; 80: 742– 751 [CrossRef] [PubMed]
    [Google Scholar]
  31. O'Toole GA, Kolter R. Flagellar and twitching motility are necessary for Pseudomonas aeruginosa biofilm development. Mol Microbiol 1998; 30: 295– 304 [CrossRef] [PubMed]
    [Google Scholar]
  32. Cramton SE, Gerke C, Götz F. In vitro methods to study staphylococcal biofilm formation. Methods Enzymol 2001; 336: 239– 255 [PubMed] [Crossref]
    [Google Scholar]
  33. Cramton SE, Ulrich M, Götz F, Döring G. Anaerobic conditions induce expression of polysaccharide intercellular adhesin in Staphylococcus aureus and Staphylococcus epidermidis. Infect Immun 2001; 69: 4079– 4085 [CrossRef] [PubMed]
    [Google Scholar]
  34. Laverty G, Gorman SP, Gilmore BF. Biomolecular mechanisms of staphylococcal biofilm formation. Future Microbiol 2013; 8: 509– 524 [CrossRef] [PubMed]
    [Google Scholar]
  35. Deighton MA, Balkau B. Adherence measured by microtiter assay as a virulence marker for Staphylococcus epidermidis infections. J Clin Microbiol 1990; 28: 2442– 2447 [PubMed]
    [Google Scholar]
  36. McCann MT, Gilmore BF, Gorman SP. Staphylococcus epidermidis device-related infections: pathogenesis and clinical management. J Pharm Pharmacol 2008; 60: 1551– 1571 [CrossRef] [PubMed]
    [Google Scholar]
  37. Hussain M, Herrmann M, von Eiff C, Perdreau-Remington F, Peters G. A 140-kilodalton extracellular protein is essential for the accumulation of Staphylococcus epidermidis strains on surfaces. Infect Immun 1997; 65: 519– 524 [PubMed]
    [Google Scholar]
  38. Boles BR, Horswill AR. Agr-mediated dispersal of Staphylococcus aureus biofilms. PLoS Pathog 2008; 4: e1000052 [CrossRef] [PubMed]
    [Google Scholar]
  39. McKenney D, Hübner J, Muller E, Wang Y, Goldmann DA et al. The ica locus of Staphylococcus epidermidis encodes production of the capsular polysaccharide/adhesin. Infect Immun 1998; 66: 4711– 4720 [PubMed]
    [Google Scholar]
  40. Rupp ME, Fey PD, Heilmann C, Götz F. Characterization of the importance of Staphylococcus epidermidis autolysin and polysaccharide intercellular adhesin in the pathogenesis of intravascular catheter-associated infection in a rat model. J Infect Dis 2001; 183: 1038– 1042 [CrossRef] [PubMed]
    [Google Scholar]
  41. Hennig S, Nyunt Wai S, Ziebuhr W. Spontaneous switch to PIA-independent biofilm formation in an ica-positive Staphylococcus epidermidis isolate. Int J Med Microbiol 2007; 297: 117– 122 [CrossRef] [PubMed]
    [Google Scholar]
  42. Cramton SE, Gerke C, Schnell NF, Nichols WW, Götz F. The intercellular adhesion (ica) locus is present in Staphylococcus aureus and is required for biofilm formation. Infect Immun 1999; 67: 5427– 5433 [PubMed]
    [Google Scholar]
  43. Kozitskaya S, Cho SH, Dietrich K, Marre R, Naber K et al. The bacterial insertion sequence element IS256 occurs preferentially in nosocomial Staphylococcus epidermidis isolates: association with biofilm formation and resistance to aminoglycosides. Infect Immun 2004; 72: 1210– 1215 [CrossRef] [PubMed]
    [Google Scholar]
  44. Fitzpatrick F, Humphreys H, O'Gara JP. Evidence for icaADBC-independent biofilm development mechanism in methicillin-resistant Staphylococcus aureus clinical isolates. J Clin Microbiol 2005; 43: 1973– 1976 [CrossRef] [PubMed]
    [Google Scholar]
  45. Shanks RM, Donegan NP, Graber ML, Buckingham SE, Zegans ME et al. Heparin stimulates Staphylococcus aureus biofilm formation. Infect Immun 2005; 73: 4596– 4606 [CrossRef] [PubMed]
    [Google Scholar]
  46. Toledo-Arana A, Merino N, Vergara-Irigaray M, Débarbouillé M, Penadés JR et al. Staphylococcus aureus develops an alternative, ica-independent biofilm in the absence of the arlRS two-component system. J Bacteriol 2005; 187: 5318– 5329 [CrossRef] [PubMed]
    [Google Scholar]
  47. Qin Z, Yang X, Yang L, Jiang J, Ou Y et al. Formation and properties of in vitro biofilms of ica-negative Staphylococcus epidermidis clinical isolates. J Med Microbiol 2007; 56: 83– 93 [CrossRef] [PubMed]
    [Google Scholar]
  48. Allesen-Holm M, Barken KB, Yang L, Klausen M, Webb JS et al. A characterization of DNA release in Pseudomonas aeruginosa cultures and biofilms. Mol Microbiol 2006; 59: 1114– 1128 [CrossRef] [PubMed]
    [Google Scholar]
  49. Whitchurch CB, Tolker-Nielsen T, Ragas PC, Mattick JS. Extracellular DNA required for bacterial biofilm formation. Science 2002; 295: 1487 [CrossRef] [PubMed]
    [Google Scholar]
  50. Talsma SS. Biofilms on medical devices. Home Healthc Nurse 2007; 25: 589– 594 [CrossRef] [PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.000466
Loading
/content/journal/jmm/10.1099/jmm.0.000466
Loading

Data & Media loading...

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