Api 50 ch strips

All isolation procedures were performed under an anaerobic environment. A cattle rumen sample was used as a bacterial source. The inner surface of the cattle rumen was sliced and chopped, and the bacteria on the rumen samples were extracted into the sterilized 10 % (v/v) glycerol solution by vigorous vortex mixing. The extracted bacterial samples were inoculated in the selection media for the isolation of hexanoic acid producer and cultured at 37 °C in a standing culture. After 7 days of cultivation, the enriched broths were transferred to the fresh selection media, and this procedure was repeated successively ten times. Then, the last enriched broths were inoculated in the fresh selection media not containing hexanoic acid and were cultured for 3 days. After confirming the presence of hexanoic acid at the final culture broth, the culture broth was serially diluted with sterilized saline solutions and spread on RCM solid plates. The plates were incubated for 7 days in an anaerobic chamber. Bacterial colonies grown on the RCM plates were serially sub-cultured to fresh plates to acquire a pure bacterial strain. Carbohydrate usage of the isolate was evaluated using API 50 CH strips (bioMérieux, France) according to the manufacturer’s instructions.

The ability of B. coagulans CGI314 to use different carbon sources was investigated with API 50CH strips (BioMérieux, Hampshire, UK), while hydrolytic activities were determined using the API-ZYM kit system (BioMérieux, Hampshire, UK) according to the instructions provided by the manufacturer.

Biochemical tests were performed using API ZYM, API 20A, and API 50CH strips (bioMérieux) according to the manufacturer's instructions. The strips were incubated for 4, 24, and 48 hr respectively.
Cellular fatty acid methyl ester (FAME) analysis was performed using Gas Chromatography/Mass Spectrometry (GC/MS). Strain Marseille‐P2341^T was grown on Columbia agar enriched with 5% sheep's blood (bioMérieux). Two samples were then prepared with approximately 50 mg of bacterial biomass per tube harvested from several culture plates. Fatty acid methyl esters were prepared as described by Sasser (Sasser, 2006). GC/MS analyses were carried out as previously described (Dione et al., 2016). In brief, fatty acid methyl esters were separated using an Elite 5‐MS column and monitored by mass spectrometry (Clarus 500—SQ 8 S, Perkin Elmer, Courtaboeuf, France). A spectral database search was performed using MS Search 2.0 operated with the Standard Reference Database 1A (NIST, Gaithersburg, USA) and the FAMEs mass spectral database (Wiley, Chichester, UK).
Antibiotic susceptibility was tested using the disc diffusion method (Le Page et al., 2015). The results were read using Scan 1200 (Interscience, Saint‐Nom‐la‐Bretèche, France).