Interplay of antibiotics and bacterial inoculum on suture-associated biofilms

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Abstract

Background: Biofilms are often antibiotic resistant, and it is unclear if prophylactic antibiotics can effectively prevent biofilm formation. Experiments were designed to test the ability of high (bactericidal) concentrations of ampicillin (AMP), vancomycin (VAN), and oxacillin (OXA) to prevent formation of suture-associated biofilms initiated with low (104) and high (10 7) numbers of Staphylococcus aureus. Materials and methods: S. aureus biofilms were cultivated overnight on silk suture incubated in biofilm growth medium supplemented with bactericidal concentrations of AMP, VAN, or OXA. Standard microbiological methods were used to quantify total numbers of viable suture-associated S. aureus. Crystal violet staining followed by spectroscopy was used to quantify biofilm biomass, which includes bacterial cells plus matrix components. To observe the effects of antibiotics on the microscopic appearance of biofilm formation, biofilms were cultivated on glass slides, then stained with fluorescent dyes, and observed by confocal microscopy. Results: In the presence of a relatively low inoculum (104) of S. aureus cells, bactericidal concentrations of AMP, VAN, or OXA were effective in preventing development of suture-associated biofilms. However, similar concentrations of these antibiotics were typically ineffective in preventing biofilm development on sutures inoculated with 107 S. aureus, a concentration relevant to contaminated skin. Confocal microscopy confirmed that bactericidal concentrations of AMP, VAN, or OXA inhibited, but did not prevent, development of S. aureus biofilms. Conclusion: Bactericidal concentrations of AMP, VAN, or OXA inhibited formation of suture-associated biofilms initiated with low numbers (104), but not high numbers (107), of S. aureus cells.

Original languageEnglish (US)
Pages (from-to)334-340
Number of pages7
JournalJournal of Surgical Research
Volume177
Issue number2
DOIs
StatePublished - Oct 2012

Bibliographical note

Funding Information:
This work was supported in part by U.S. National Institutes of Health Grant R01 GM095553 (to CW) and in part by funds from the Department of Surgery, University of Minnesota, Minneapolis (to DH). Parts of this work were performed in the Institute of Technology Characterization Facility, University of Minnesota, which receives partial support from NSF through the MRSEC program.

Keywords

  • Ampicillin
  • Antibiotics
  • Biofilm
  • Biomass
  • Confocal microscopy
  • Oxacillin
  • Staphylococcus aureus
  • Suture
  • Vancomycin

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