Omnilog software

Selected strains were identified on a Biolog GEN III (Biolog, Hayward, CA, USA) microtiter plate, containing 71 different carbon sources and 23 chemical sensitivity assays. After 24-h incubation on TSA plates, single colonies were transferred into inoculating fluid. The bacterial suspension was adjusted to 95% transmittance using a Biolog turbidimeter. Then, 150 μL of the suspension was dispensed into each well of the Biolog GEN III microplate. Inoculated plates were monitored via Biolog’s OmniLog software for 48 h at 37 °C with measurements taken every 15 min. Their identity was confirmed using the Biolog GEN III MicroPlate, based on phenotypic similarity to the control strains in the BIOLOG database.

A high-throughput analysis for microbial growth phenotypes using a colorimetric reaction, Phenotype MicroArrays (Biolog Inc., Hayward, CA, USA), was performed in accordance with the manufacturer’s protocol. Briefly, several colonies of E. coli grown on LB agar were transferred in 10 mL of a mixture of Biolog IF-0a medium (Biolog) and sterilized water into a sterile capped test tube. The suspension was mixed gently, and the turbidity was adjusted to achieve the appropriate transmittance using the Biolog turbidimeter (Biolog). The cell suspension was diluted with the IF-0a Plus dye mix, as mentioned in the manufacturer’s protocol. One hundred microliters of the mixture suspension was inoculated into phenotypic microarray (PM) plates 1 to 3 and 9 to 20 and incubated for 72 h at 37°C. The absorbance of each well was taken every 15 min. The OmniLog software (Biolog) was used to view and edit data, to compare data lists, and to generate reports.