Petri dish

Sensitivity to antibiotics was determined on a Mueller–Hinton agar in a petri dish (BioMerieux, Marcy-l'Etoile, France). The following antibiotics were tested using Sirscan discs (i2a, Perols, France): doxycycline, rifampicin, vancomycin, amoxicillin, erythromycin, ceftriaxone, ciprofloxacin, gentamicin, penicillin, trimethoprim/sulfamethoxazole, imipenem, and metronidazole. Scan 1200 was used to interpret the results (Interscience, Saint Nom la Bretêche, France).
The commercially available API ZYM, API 50CH, and API 20 NE strips (BioMerieux, Marcy-l'Etoile, France) were used for biochemical tests according to the manufacturer's instructions. The time of incubation was 4 h for API ZYM and 48 h for the others.
Growth of the strain Awa-1^T was tested with different growth temperatures (25°C, 30°C, 37°C, 45°C) under aerobic conditions and also in anaerobic and microaerophilic atmospheres, created using AnaeroGen^TM (Atmosphere Generation Systems, Dardily, France) and anaerobic jars (Mitsubishi) with GENbag microaer system (BioMerieux), respectively.

Gram staining and motility were observed from fresh colonies between blades and slats using a DM1000 photonic microscope (Leica Microsystems, Nanterre, France) with a 40 × objective lens [16] (link). Spore formation was determined by thermal shock (80°C for 20 minutes) and observed under a microscope. Negative staining was carried out with detection formvar-coated grids placed on a drop of 40 μL of bacterial suspension and incubated at 37°C for 30 minutes, followed by a 10-second incubation in 1% ammonium molybdate. The grids were dried on blotting paper and then observed with a Tecnai G20 transmission electron microscope (FEI Company, Limeil-Brévannes, France). We studied the biochemical characteristics of this strain using API 20NE, API ZYM and API 50CH strips according to the manufacturer's instructions (bioMérieux). Oxidase and catalase reactions were determined using a BBL DrySlide (Becton, Le Pont de Claix, France) according to the manufacturer's instructions. The antimicrobial activity test was performed using the disc diffusion method (i2a, Montpellier, France) [17] (link) on Mueller-Hinton agar in a petri dish (bioMérieux).

Biochemical tests were performed using the commercially available Api ZYM (bioMérieux, Marcy l’Étoile, France), API 50CH (bioMérieux) and 20 NE (bioMérieux) strips. The incubation time was 48 hours for the API 50CH and 20 NE, and 4 hours for Api ZYM. Growth of strain SK-1T was tested in aerobic atmosphere, in the presence of 5% CO₂ and also in anaerobic and microaerophilic atmospheres, created using AnaeroGen (Atmosphere Generation Systems, Dardily, France). Antibiotic susceptibility was determined by Müller-Hinton agar in a petri dish (bioMérieux). The following antibiotics were tested: doxycycline, rifampicin, vancomycin, nitrofurantoin, amoxicillin, erythromycin, ampicillin, ceftriaxone, ciprofloxacin, gentamicin, penicillin, trimethoprim/sulfamethoxazole, imipenem and metronidazole.