Bactericidal effect of ϕAB2 in a liquid suspension To determine the bactericidal effect Dactolisib supplier of ϕAB2 in suspension, A. baumannii M3237 was Y-27632 mw cultured overnight and then transferred to a flask and incubated at 37°C until reaching an OD600 of 1.0 (5 × 108 CFU/ml). A. baumannii M3237 cultures were then serially diluted to obtain final concentrations of 5 × 106, 5 × 105 or 5 × 104 CFU/ml. A 1 ml aliquot of each concentration was mixed with 1 ml of ϕAB2 suspension to obtain a final phage

concentration of 103, 105, or 108 PFU/ml. Phage-free culture (containing bacteria only) was included as a control. Following a 5- or 10-min incubation, host and phage mixtures were immediately passed through 47-mm diameter membrane filters (pore size of 0.45 μm, Pall Corporation) and washed with 20 ml of phosphate-buffered saline (PBS) to remove unattached phages [26]. PHA-848125 Washed filters were placed in separate dishes containing LB agar, and following 24-h incubation at 37°C, the number of recovered A. baumannii M3237 was calculated by counting colonies on each filter. The survival rate was calculated as log10 of Nt/N0, where N0 is the number of A. baumannii M3237 colonies recovered on the control filter and Nt

is the number of colonies on the test filter. Bactericidal effect of ϕAB2 on a glass slide To determine the bactericidal effect of ϕAB2 on a glass surface, glass slides were sterilized, pre-contaminated with A. baumannii M3237 by spreading diluted culture stock solution on the glass surface to obtain concentrations of 104, 105, and 106 CFU/slide and dried for 30 min in a biosafety hood at room temperature. Then, slides were divided into two groups. 1) test: treated with ϕAB2 to reach a concentration stiripentol of 103, 105, or 108 PFU/slide and 2) control: treated with phage-free suspension. After the ϕAB2 solution or phage-free suspension was applied to the A. baumannii M3237 slide, they were stored for 5 or 10 min at room temperature. Residual A. baumannii M3237 particles on the test or control slides were eluted with 20 ml of peptone into a conical tube, gently vortexed for 30 s, serially

diluted and passed through membrane filters, as above. The filters were then washed with PBS, placed on LB agar plates, and incubated for 24 h at 37°C. The number of A. baumannii M3237 colonies that grew on each filter was counted and the survival rate was calculated. Production of ϕAB2 hand sanitizer in a paraffin oil-based lotion A commercial cream containing paraffin mineral oil (First Chemical Works, Taipei, Taiwan) was combined with ϕAB2 in a conical tube and sterile water added to obtain a paraffin oil-based lotion with a final concentration of 10% (v/v) paraffin oil and a phage concentration of 108 PFU/ml. The phage-containing lotion was stored at room temperature up to 30 days. At each sampling point, the phage lotion was inoculated for plaque assays to obtain a kinetic curve of the phage concentration.