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The diagnosis of implant-associated infections is hampered due to microbial adherence and biofilm formation on the implant surface. Sonication of explanted devices was shown to improve the microbiological diagnosis by physical removal of biofilms. Recently, chemical agents have been investigated for biofilm dislodgement such as the chelating agent ethylenediaminetetraacetic acid (EDTA) and the reducing agent dithiothreitol (DTT). We compared the activity of chemical methods for biofilm dislodgement to sonication in an established  in vitro  model of artificial biofilm. Biofilm-producing laboratory strains of  Staphylococcus epidermidis  (ATCC 35984),  S .  aureus  (ATCC 43300),  E .  coli  (ATCC 25922) and  Pseudomonas aeruginosa  (ATCC 53278) were used. After 3 days of biofilm formation, porous glass beads were exposed to control (0.9% NaCl), sonication or chemical agents. Quantitative and qualitative biofilm analyses were performed by colony counting, isothermal microcalorimetry and scanning electron microscopy. Recovered colony counts after treatment with EDTA and DTT were similar to those after exposure to 0.9% NaCl for biofilms of  S .  epidermidis  (6.3 and 6.1 vs. 6.0 log 10  CFU/mL,  S .  aureus  (6.4 and 6.3 vs. 6.3 log 10  CFU/mL),  E .  coli  (5.2 and 5.1 vs. 5.1 log 10  CFU/mL and  P .  aeruginosa  (5.1 and 5.2 vs. 5.0 log 10  CFU/mL, respectively). In contrast, with sonication higher CFU counts were detected with all tested microorganisms (7.5, 7.3, 6.2 and 6.5 log 10  CFU/mL, respectively) (p <0.05). Concordant results were observed with isothermal microcalorimetry and scanning electron microscopy. In conclusion, sonication is superior to both tested chemical methods (EDTA and DTT) for dislodgement of  S .  epidermidis ,  S .  aureus ,  E .  coli  and  P .  aeruginosa  biofilms. Future studies may evaluate potential additive effect of chemical dislodgement to sonication.

plos one

Comparison of sonication with chemical biofilm dislodgement methods using chelating and reducing agents: Implications for the microbiological diagnosis of implant associated infection