Biofilm incubation on stainless steel beads

SS Sumbul Shaikh
NL Norman A. Lapin
BP Bibin Prasad
CS Carolyn R. Sturge
CP Christine Pybus
RP Reed Pifer
QW Qi Wang
BE Bret M. Evers
RC Rajiv Chopra
DG David E. Greenberg
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Biofilm was grown on stainless steel beads (6mm) using Staphylococcus aureus U1 (Lux UAMS-1; MSSA; generously provided by Dr. Mark Smeltzer) or Pseudomonas aeruginosa (Lux-PAO1, ATCC, Manassas, VA). An isolated colony was inoculated into 3 ml of Mueller–Hinton broth (MHII; Fisher Scientific), and incubated overnight at 37 °C, at 220 RPM. A bacterial suspension of 1 × 106 CFU/ml was created from this culture. Biofilm was produced by placing a 6mm stainless steel bead in a conical tube containing 4 ml of the bacterial suspension and incubated in a shaking incubator at 110 RPM for 2 h at 37 °C. The initial log10CFU bacterial count on the un-implanted balls for S. aureus and P. aeruginosa were on average 5.0 and 4.6, respectively.

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