Figure 1 Phylogenetic tree highlighting the position of Alistipes senegalensis strain JC50T relative to other type strains within the Alistipes genus. GenBank accession numbers are indicated in parentheses. The tree was inferred from the comparison of 16S rRNA … Different growth temperatures (25, 30, 37, 45��C) were tested; no growth occurred at 25��C and especially 45��C, growth occurred between 30 and 37��C, and optimal growth was observed at 37��C. Colonies were 0.2 to 0.3 mm in diameter on blood-enriched Columbia agar and Brain Heart Infusion (BHI) agar. Growth of the strain was tested under anaerobic and microaerophilic conditions using GENbag anaer and GENbag microaer systems, respectively (BioM��rieux), and in the presence of air, with or without 5% CO2 and in aerobic conditions.

The optimal growth of the strain was obtained anaerobically, with weak growth being observed under microaerophilic conditions, and no growth observed under aerobic conditions. Gram staining showed Gram negative bacilli. A motility test was negative. Cells grown on agar are Gram-negative rod-shaped bacteria (Figure 2) and have a mean diameter of 0.56 ��m (Figure 3) by electron microscopy. Figure 2 Gram staining of A. senegalensis strain JC50T Figure 3 Transmission electron microscopy of A. senegalensis strain JC50T, using a Morgani 268D (Philips) at an operating voltage of 60kV.The scale bar represents 900 nm. Strain JC50T exhibited a catalase activity but no oxidase activity. Using API Rapid ID 32A, a positive reaction was obtained for mannose fermentation, proline arylimidase, leucyl glycine arylamidase, alanine arylamidase.

A weak reaction was obtained for indole production, ��-galactosidase, ��-galactosidase, ��-glucuronidase, arginine arlyamidase and glycine arylamidase. A. senegalensis is susceptible to penicillin G, imipeneme, amoxicillin + clavulanic acid and clindamycin but resistant to metronidazole and vancomycin. Matrix-assisted laser-desorption/ionization time-of-flight (MALDI-TOF) MS protein analysis was carried out as previously described [20]. Briefly, a pipette tip was used to pick one isolated bacterial colony from a culture agar plate, and spread as a thin film on an MTP 384 MALDI-TOF target plate (Bruker Daltonics, Germany). Four distinct deposits were done for strain JC50T from four isolated colonies.

Each smear was overlaid with 2��L of matrix solution (saturated solution of alpha-cyano-4-hydroxycinnamic acid) in 50% acetonitrile, 2.5% tri-fluoracetic acid, and allowed to dry for five minutes. Measurements were performed with a Microflex spectrometer (Bruker). Spectra were recorded in the positive linear mode for the mass range of 2,000 to 20,000 Da (parameter settings: ion source 1 (ISI), 20kV; IS2, 18.5 kV; lens, 7 kV). A spectrum was obtained after 675 shots at a variable laser Dacomitinib power.