Api 50ch systems

The biochemical reactions toward various substrates were determined by API 20NE and API 50CH systems (bioMérieux), according to the manufacturers’ instructions. Enzyme activities and other biochemical properties were conducted by API ZYM assay (bioMérieux). The biochemical phenotypes of strain CHPC 1.3453^T were compared with those of three type trains, B. diminuta ATCC 11568^T, B. vesicularis NBRC 12165^T, and B. halotolerans MCS24^T, in each independent experiment.

The six strains were phenotypically characterized in detail. Bacteria were streaked on Columbia agar with 5% defibrinated sheep blood and incubated at 25°C under aerobic condition for 2 days to observe the color of the colonies. Growth response to different temperatures (25, 37, and 42°C) were determined by spreading bacterial cells of 0.5 McFarland on Columbia agar with 5% defibrinated sheep blood agar followed by incubation under aerobic condition for up to 7 days. Hydrolysis of tyrosine and xanthine were tested as described previously (Conville and Witebsky, 2007 ). Biochemical data were obtained using the API 50 CH systems (bioMérieux, France) at 25°C according to manufacturers’ instructions. Reading intervals and durations of the tests followed the protocol suggested by Kattar et al. (2001) (link). Cellular fatty acids were extracted directly from lyophilized cells (approximately 30 mg of original wet weight) grown on Columbia agar with 5% defibrinated sheep blood at 25°C for 2 days and analyzed by gas chromatography as described by Microbial ID, Inc. (Sasser, 1990 ) with peak-naming performed using MIDI, Inc. Sherlock Rapid Libraries and Methods.