Optimal growth was obtained anaerobically, with weak growth being observed in microaerophilic condition, and no growth occurring in aerobic conditions and with 5% CO2. Gram staining showed Gram positive cocci. A motility test was negative. Cells grown on agar are read FAQ Gram-positive (Figure 2) and have a mean diameter of 0.71 ��m by electron microscopy and are mostly grouped in pairs, short chains or small clumps (Figure 3). Figure 2 Gram staining of A. vaginalis strain PH9 Figure 3 Transmission electron microscopy of A. vaginalis strain PH9, using a Morgani 268D (Philips) at an operating voltage of 60kV.The scale bar represents 900 nm. Strain PH9 exhibited catalase activity but no oxidase activity. Using API Rapid ID 32A, a positive reaction was observed for arginine dihydrolase, histidine arylamidase, leucine arylamidase and mannose fermentation.

A weak activity was observed for glycine arylamidase. A. vaginalis is susceptible to penicillin G, imipeneme, amoxicillin + clavulanic acid, vancomycin, clindamycin and metronidazole. Matrix-assisted laser-desorption/ionization time-of-flight (MALDI-TOF) MS protein analysis was carried out as previously described [19]. Briefly, a pipette tip was used to pick one isolated bacterial colony from a culture agar plate, and to spread it as a thin film on a MTP 384 MALDI-TOF target plate (Bruker Daltonics, Germany). Twelve distinct deposits were done for strain PH9 from twelve 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 power. The time of acquisition was between 30 seconds and 1 minute per spot. The twelve PH9 spectra were imported into the MALDI Bio Typer software (version 2.0, Bruker) and analyzed by standard pattern matching (with default parameter settings) against the main spectra of 2,843 bacteria, including spectra from seven validated Anaerococcus species used as reference data, in the Bio Typer database. The method of identification includes the m/z from 3,000 to 15,000 Da.

For every spectrum, Entinostat 100 peaks at most were taken into account and compared with the spectra in the database. A score enabled the presumptive identification and discrimination of the tested species from those in the database: a score �� 2 with a validated species enabled the identification at the species level; a score �� 1.7 but < 2 enabled the identification at the genus level; and a score < 1.7 did not enable any identification. Spectra were compared with the Bruker database that contained spectra from the seven validated Anaerococcus species.