GenBank accession numbers are indicated in parentheses. Sequences were aligned using CLUSTALW, and free copy … Different growth temperatures (25, 30, 37, 45��C) were tested. No growth occurred at either 25��C or 45��C, growth occurred at either 30 or 37��C. Optimal growth was observed at 37��C. Colonies were light brown, opaque and 0.5 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, of 5% CO2 and in anaerobic conditions. Optimal growth was obtained aerobically, with weak growth being observed under microaerophilic condition and with 5% CO2. No growth occurred under anaerobic conditions.

Gram staining showed Gram negative curved rods (Figure 2). A motility test was positive. Cells grown on agar have a mean diameter of 0.44 ��m by electron microscopy and have several polar flagella (Figure 3). Figure 2 Gram staining of H. massiliense strain JC206T. Figure 3 Transmission electron microscopy of H. massiliense strain JC206T, using a Morgani 268D (Philips) at an operating voltage of 60kV.The scale bar represents 900 nm. Strain 206T exhibited catalase and oxidase activities. Using an API 20 NE strip (BioMerieux), nitrate reduction, indole formation, glucose fermentation and urease were negative. Arginine dihydrolase and esculin hydrolysis were positive. H. massiliense is susceptible to ticarcillin, imipenem, trimethoprim/sulfamethoxazole, gentamicin, amikacin, and colimycin but resistant to fosfomycin and nitrofurantoin.

Matrix-assisted laser-desorption/ionization time-of-flight (MALDI-TOF) MS protein analysis was carried out as previously described [30] using a Microflex spectrometer (Bruker Daltonics, Germany). Spectra were compared with the Bruker database that contained no spectrum from Herbaspirillum Dacomitinib species. No significant score was obtained with any other taxon. We incremented our database with the spectrum from strain JC206 T (Figure 4). Figure 4 Reference mass spectrum from H. massiliense strain JC206T. Spectra from 12 individual colonies were compared and a reference spectrum was generated. Genome sequencing information Genome project history The organism was selected for sequencing on the basis of its phylogenetic position and 16S rRNA similarity to other members of the genus Herbaspirillum, and is part of a ��culturomics�� study of the human digestive flora aiming at isolating all bacterial species within human feces. It was the second genome of a Herbaspirillum species and the first genome of H. massiliense sp. nov. A summary of the project information is shown in Table 2.