Bacterial species representative of the spectrum of microbial keratitis causative Gram-positive and Gram-negative pathogens (Staphylococcus aureus, Klebsiella pneumoniae and Enterococcus avium (previously classified as group D Streptococcus)) were used to define the mock bacterial community. Klebsiella pneumoniae and Staphylococcus aureus were grown overnight at 37 °C in lysogeny broth (LB) whilst Staphylococcus aureus was grown in Brain Heart Infusion (BHI) broth. Negative controls consisted of the LB and BHI broth without any inoculum, respectively. Overnight cultures were diluted into fresh medium to an optical density 600 nm (OD600) of 0.05 and incubated at 37 °C with aeration. The culture OD600 was measured every 30 min for 5 h using spectrophotometer (Ultrospec 2100 pro, Amersham Biosciences, UK). For enumeration of bacteria, cultures were plated onto agar plates (Enterococcus avium, blood agar; Staphylococcus aureus and Klebsiella pneumoniae, LB agar respectively) and incubated overnight at 37 °C. The colony-forming units (CFUs) for each species were enumerated the following day. Bacterial growth curves were plotted and the mid-exponential growth phases of the samples were taken. The mock bacterial community consisted of 1 × 105 CFU/ml each of Klebsiella pneumoniae, Enterococcus avium and Staphylococcus aureus.

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