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Antibiotic susceptibility for P. aeruginosa P16, E. coli DH5α harboring an empty vector and vector expressing OXA-205 was determined in Mueller Hinton broth (Liofilchem), using a final inoculum of 5 × 10 CFU/ml.
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A 10 μl aliquot of the microorganism was used to inoculate each microtiter plate well to achieve a final inoculum size of 1 × 10 CFU/ml.
A 10 μl aliquot of the standard laboratory strain of E. coli ATCC 8739 was used to inoculate each microtiter plate well to achieve a final inoculum size of 5 × 105 CFU/ml.
A final inoculum of ~5 × 105 CFU/ml was used for all in-vitro experiments.
The MIC for M. tuberculosis H37Rv and drug resistant clinical sample of M. tuberculosis was determined using a broth microdilution method in Middlebrook 7H9 medium supplemented with OADC, with a final inoculum of 5 × 102 cells/mL.
A final inoculum of 5 × 105 CFU/ml in Mueller-Hinton broth containing 2 5% lysed horse blood was used to inoculate the Sensititre STPF3 standard plate (Trek Diagnostics, East Essex, England).
Bacterial suspensions were diluted in sterile Muller Hington Broth (MHB) to obtain a final inoculum of 106 CFU/ ml.
Sixteen mice were inoculated with a 1∶1 mix of PA14 and PA14glpT::MAR2xT7 for a final inoculum size approximately of 5×105.
The bacterial suspension was then added to each well to provide a final inoculum density of 5×105 cfu/ml.
For the low-inoculum experiments, a further 1-in-4,000 dilution was made in Ca-MH broth to achieve a final inoculum of ∼4 ×10 CFU/ml.
Conidial suspensions were further diluted 1 10 in medium for a final inoculum concentration of 1.0 x 10 CFU/mL.
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