Gram staining, motility, catalase and oxidase were determined as described by Lagier et al.[3] (link). Sporulation was tested using a thermal shock on bacterial colonies (diluted in phosphate-buffered saline) for 10 minutes at 80°C. The biochemical characteristics were tested using API 50CH, API ZYM and API 20A strips (bioMérieux). Antibiotic susceptibility referred to European Committee on Antimicrobial Susceptibility Testing 2015 recommendations.
Api 20a strips
Manufactured by bioMérieux
Sourced in France
The API 20A strips are a standardized identification system for anaerobic bacteria, designed for use in clinical microbiology laboratories. The strips contain 20 microtubes filled with dehydrated biochemical substrates that allow for the identification of a wide range of anaerobic bacterial species.
Automatically generated - may contain errors
2 protocols using api 20a strips
1
Bacterial Phenotypic Characterization
2
Isolation and Antibiotic Resistance of Clostridium baratii
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C. baratii C3 was routinely cultured in Reinforced Clostridium Medium (RCM, Becton-Dickinson, Franklin Lakes, NJ, USA) supplemented with 0.5 g/L of L-cysteine (RCM-cys; Loba Chemie, India). All incubations were performed at 37 °C for 24–48 h in an anaerobic jar (Anaerocult, Merck, Darmstadt, Germany) with anaerobic packs (Gaspak EM, Becton-Dickinson, Franklin Lakes, NJ, USA). Two overnight cultures of C. baratii were made on Blood Agar and Starch Agar plates (Winkler, Santiago, Chile). The bacteria were also inoculated in API-20A strips (Biomerieux, Craponne, France) following the manufacturer’s instructions. Finally, antibiotic resistance patterns were evaluated using antibiotic discs in RCM agar plates incubated for 48 h (Lilfilchem, Roseto degli Abruzzi TE, Italy) for 48 h. Cefazolin (30 µg), ciprofloxacin (5 µg), chloramphenicol (30 µg), erythromycin (15 µg), penicillin G (10 µg), piperacill/tazobactam (110 µg), trimethoprim-sulfamethoxazole (25 µg), and vancomycin (30 µg) were used.
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