culture was then incubated at 37uC for 1224 hours. The diameter of a zone of inhibition was measured which is a qualitative measure of the inhibitory activity of a compound. The data represents the distances from the edge of the disks to the end of the clear zone, where growth begins. Each experiment was repeated at least three times. doi:10.1371/journal.pone.0033777.t003 String and Precipitation test for Hypermucoviscosity The NTUH-K2044 and NTUH-K2044DcpxAR was streaked onto LB agar plates and incubated at 37uC overnight. A standard bacteriologic loop was used to stretch ” a mucoviscous string from the colony. Hypermucoviscosity was defined by the formation of viscous strings.5 mm in length when a loop was used to stretch the colony on agar plate which was considered the positive string test. The strains to be tested were cultured overnight in LB broth at 37uC and subjected to centrifugation at 1,0006g for 5 min to 10595516” check reduction in mucoidy. For exopolysaccharide analysis, cells were grown to late log phase in shaking culture and stained with crystal violet followed by treatment with 20% copper sulphate solution. Samples were visualized using an Olympus microscope work station. Capsular polysaccharides were extracted from overnight bacterial suspensions adjusted to,108 cells per ml with Zwittergent 314 detergent. The amount of uronic acid was then measured according to the method described previously. Each experiment was performed in triplicate. Antibiotic susceptibility testing Strains in this study were examined for resistance to nalidixic acid: NA30, colistin: CL30, enrofloxacin: EX10, polymyxin B: PB300, ciprofloxacin: CF5, azithromycin: AT15, erythromycin: E15, tetracycline: T30, rifampicin: R5, trimethoprim: TR5, kanamycin: K30, streptomycin: S10, tobramycin: TB10, clindamycin: CD2, spectinomycin: S100, imipenem: I10, ampicillin: A10, ertapenem: ETP10, piperacillin: PC100, ticarcillin: TI75, ceftazidime: CA30, chloramphenicol: C30, ceftriaxone: CI30, cefepime: CPM30 and carbencillin: CB100 by using commercial discs as described previously according to the interpretation criteria recommended by Clinical and Laboratory Standards Institute CLSI. MIC of antibiotics was tested using E-strips. Interpretation was done as per the criteria approved by CLSI. E. coli ATCC 25922 was used as a reference strain as recommended. Scanning electron microscopy Overnight cultures were fixed after harvesting; cells were washed three times with ice-cold NaCl/Pi. The cells were then resuspended in NaCl/Pi, adhered to cover slips that had been coated with 0.1% poly. Adherent cells were washed with NaCl/Pi and then dehydrated using an ascending series of ethanol incubations. Finally, cells on covers lips were infiltrated with tbutyl alcohol and freeze-dried in a lyophilizer. Dried samples were sputter-coated with gold/palladium and then observed under a scanning electron microscope. In vitro growth curves To examine bacterial growth in vitro, overnight cultures were diluted 1:100 and subcultured for 10 h. The growth kinetics was monitored with LB at different pH. The growth Outer membrane proteins SCH 58261 supplier preparation Outer membrane proteins were purified by the method as described previously. Cells were harvested by centrifugation CpxAR Confers b-Lactam Resistance and were suspended in 50 mM Tris-HCl buffer containing 5 mM phenylmethylsulfonyl fluoride and sonicated for 15 mins. The crushed 
material was treated with DNase and RNase, and the unbroken cells were r
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