Distribution (%) of MIC values (in µg/mL) of group n°2 antimicrobials.

<p>MICs of marbofloxacin, danofloxacin, gamithromycin, tildipirosin, tulathromycin, valnemulin for 27 <i>M. bovis</i> strains isolated in 1978–1979 (white bars) and 30 <i>M. bovis</i> strains isolated in 2010–2012 (black bars). When available, CLSI breakpoints for bovine <i>Pasteurellaceae</i> are given under the X axis: - strains with MIC values less than or equal to the dilution indicated in the dotted-line arrow are susceptible, - strains with MIC values greater than the dilution indicated in the full-line arrow are resistant, all other strains are intermediate.</p

Distribution (%) of MIC values (in µg/mL) of group n°1 antimicrobials.

<p>MICs of tylosin, tilmicosin, spectinomycin, oxytetracycline and florfenicol for 27 <i>M. bovis</i> strains isolated in 1978–1979 (white bars) and 46 isolated in 2010–2012 (black bars). MICs of enrofloxacin, for 27 <i>M. bovis</i> strains isolated in 1978- 1979 (white bars) and 143 <i>M. bovis</i> strains isolated in 2010–2012 (black bars). When available, CLSI breakpoints for bovine <i>Pasteurellaceae</i> are given under the X axis: - strains with MIC values less than or equal to the dilution indicated in the dotted-line arrow are susceptible, - strains with MIC values greater than the dilution indicated in the full-line arrow are resistant, - all other strains are intermediate.</p

Exopolysaccharide (EPS) secretion by <i>Mmm</i> strain Afadé in supplemented PPLO broth and CMRL-1066 medium.

<p>EPS extracted either from <i>Mmm</i> culture (Afadé) or from unseeded medium used as negative control (Neg) were spotted onto a nylon membrane and either stained with a glycoprotein detection reagent (<b>A</b>) or immunodetected using a serum from a CBPP-positive bovine (<b>B</b>).</p

Variation in <i>Mmm</i> colony opacity associated to antigenic variation of the glucose PTS permease.

<p><b>A: </b><i>Mmm</i> strain Afadé colonies on PPLO supplemented agar observed under stereomicroscope with indirect light. <b>B:</b> Colony-blotting performed from the same plate using the 3F3 monoclonal antibody. (Magnification: x10).</p

PG1^T colony opacity is related to the presence of capsular material.

<p><b>A:</b> Colony morphology of PG1^T opaque (OP) and translucent (TR) colony variants grown on PPLO supplemented agar with (right) or without (left) Congo red. Magnification: x30. <b>B:</b> Electron microscopy of Ruthenium red-stained PG1^T OP and TR colonies. CM: cytoplasmic membrane.</p

Kinetics of exopolysaccharides secretion and viability of <b><i>Mmm</i></b><b> strain Afadé following transfer into CMRL-1066 medium with (red) or without (blue) glucose supplementation.</b>

<p>Glucose (1 g/L) was added at 24, 48 and 72 h (arrows).</p

NMR spectra of EPS produced by <i>Mmm</i>.

<p><b>A: ¹H NMR spectrum, B: 2D COSY NMR spectrum and C: 2D HSQC NMR spectrum.</b></p

Growth, pH and exopolysaccharide (EPS) secretion of translucent (blue) and opaque (red) colony variants in CMRL-1066 medium.

<p>50 ml of CMRL-1066 medium were inoculated with 10⁸–10⁹ CFU/mL and incubated for 96 hours. Viable titer, pH and EPS production were determined every 24 h.</p

Bayesian inference of <i>Mycoplasma mycoides</i> subsp. <i>mycoides</i> “Small Colony” evolutionary history.

<p>The maximum credibility tree resulted from BEAST analysis with the concatenated sequence of 62 core genes and using a strict molecular clock and a GTR+I substitution model. Branches are scaled by time according to the scales displayed at the bottom and top of the figure. Strain names are colored according to the sampling location (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0046821#pone-0046821-g001" target="_blank">Figure 1</a>). Country codes are indicated in brackets (for abbreviations refer to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0046821#pone-0046821-g002" target="_blank">Figure 2</a>). The dates at the nodes refer to the most recent common ancestor (MRCA) estimated by BEAST for all strains deriving from these nodes. The 95% Highest Posterior Densities (HPD) values are displayed in brackets. The node leading to the Australasian and South African strains was used as a control of estimation accuracy. The model dates their MRCA around 1850. This agrees with the dates of CBPP introduction in Australia (1858) and Southern Africa (1853), indicated in blue boxes. According to this model, the MRCA for all the tested strains emerged around 1700 and the MRCA for all the Sub-Saharan strains around 1810.</p

Phylogeny of <i>Mycoplasma mycoides</i> subsp. <i>mycoides</i> “Small Colony” (MmmSC) inferred by the maximum likelihood method.

<p>The maximum likelihood tree was reconstructed using PhyML (GTR with invariable sites) based on the alignment of 62 concatenated core genes of MmmSC. Bootstrap values >80% are shown. Strain names are colored according to the sampling location (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0046821#pone-0046821-g001" target="_blank">Figure 1</a>). Country codes are indicated in brackets. The branch corresponding to the outgroup (GM12) was shortened, as indicated by two parallel bars. The scale indicates the number of substitutions per site. Abbreviations: AU = Australia; CM = Cameroon; ES = Spain; ER = Eritrea; ET = Ethiopia; FR = France; GN = Guinea; IN = India; IT = Italy; ML = Mali; PT = Portugal; RW = Rwanda; SD = Sudan; TZ = Tanzania; ZM = Zambia.</p