Relatedness of <i>Salmonella enterica</i> ICMS spectra reflects serotype.

<p>(A) Global cluster analysis of <i>S. enterica</i> isolates. (B) Enlargement of major clusters from (A). Serovars: <i>S</i>. Typhi (red), <i>S.</i> Typhimurium (green), <i>S</i>. Enteritidis (yellow), others (blue). Isolate sources: G:Göttingen; R:Salmonella Reference Center; E:Eikwe; N:Nkawkaw; f:Fosso. Isolation time points in Ghana (E, N, and f only): not bold  = 2006; bold  = 2009 (C) Overlay of ICMS spectra contained in the four major clusters identifies at least one major peak (peak 2; m/z  = 5713.9) specific to <i>S</i>. Typhi (red) and two major peaks (peaks 1 and 3; m/z = 5616.7 and m/z = 6009.7 respectively) specific for non-<i>S</i>. Typhi isolates (green, yellow and blue). Several other small peaks specific for <i>S</i>. Typhi were also seen (three example peaks indicated in cluster IIb by arrows, m/z = 2856.4, m/z = 3258.0, and m/z = 4716.3, respectively).</p

Value of BioTyper hit scores for <i>S.</i> Typhi identification.

<p>Negative values indicated higher false hit scores, positive values indicated higher correct hit scores. A value near zero indicated a similar score distribution between correct and false hits.</p

Concordance of species identification by conventional and ICMS methods.

a<p>In the case of absence or discordance of identifications by conventional and ICMS, the correct species was identified by sequencing of the 16S rDNA locus.</p

Gentamicin protection assays.

<p>1. wild type strain NCTC 11168, 2. mutant strain NCTC 11168::<i>cj0268c</i>, 3. complemented mutant NCTC 11168::<i>cj0268c</i>-comp-<i>cj0268c</i>. Gentamicin protection assays with the strains under investigation using (a) Caco2 cells and (b) PCC-cells. The assays on Caco2 cells with the parental strain, the knockout mutant and the complemented knockout mutant confirmed the infection-deficient phenotype to be due to the functional loss of <i>cj0268c.</i> Four independent experiments have been carried out, respectively. The standard deviations are indicated. (a) Taking the number of <i>C. jejuni</i> wild type strain NCTC 11168 colonies recovered as 100%, the mean value of <i>cj0268c</i>-mutant colonies (NCTC 11168::<i>cj0268c</i>) accounted for 62% (±2.41), whereas the percentage of obtained colonies from the complemented strain (NCTC 11168::<i>cj0268c</i>-comp-<i>cj0268c)</i> was 93% (±2.04). Since the <i>P</i>-value for the mutant was less than 0.001, the reduced invasion capacity was significant. (b) The relevance of <i>cj0268c</i> for invasion is not restricted to human cells, since it could also be shown for the invasion of PCC-cells. Compared to the invasion capacity of parental strain NCTC 11168 (100%), the percentage of colonies obtained from mutant strain NCTC 11168::<i>cj0268c was</i> only 62% (±2.36, P<0.0007). However, complementation of the mutant strain with <i>cj0268c</i> restored the infectivity of NCTC 11168::<i>cj0268c</i>-comp-<i>cj0268c</i> which was similar to that of the parental strain (96%, ±4.96).</p

Flow cytometric measurements of permeabilized and non-permeabilized <i>E.</i><i> coli</i> cells expressing Cj0268c (pRRC-<i>cj0268c</i>) or transformed with an empty vector (pRRC).

<p>Bacteria were immunolabelled with a monoclonal anti-pentaHis primary antibody and a R-phycoerythrin-conjugated secondary antibody. E. coli cells of both populations were gated according to the forward scatter and the side scatter (dot plots, left panel). Permeabilized <i>E. coli</i> cells harbouring the empty plasmid pRRC without gene <i>cj0268c</i> (upper panel) possess a considerable lower fluorescence intensity as compared to an <i>E. coli</i> population which express Cj0268c (lower panel). To verify the specificity of binding of anti-pentaHis, a control staining with the secondary antibody only was included (right panel).</p

Detection of Cj0268c in <i>E. coli</i> and altered adherence.

<p>1. <i>E. coli</i> DH5α/pRRC, 2. <i>E. coli</i> DH5α/pRRC-cj0268cHis. (a) Immunoblot with a monoclonal antibody against the His-tag detected a protein with a molecular weight of 41 kDa corresponding to Cj0268c. The protein was exclusively found in the lysate of the Cj0268c-expressing <i>E. coli</i>. The respective Coomassie staining served as a loading control. (b) Adherence assays on Caco2 cells with Cj0268c-expressing <i>E. coli</i> and <i>E. coli</i> transformed with plasmid pRRC alone. Taken the number of recovered <i>E. coli</i> colonies from the strain harbouring pRRC without <i>cj0268c</i> as 100%, the mean value of the adherence capacity of <i>E. coli</i> DH5α/pRRC-cj0268cHis was 201.7% (±6.44, P<0.0039).</p

Resistance to Triton X-100.

<p>(1) Parental strain NCTC 11168, (2) <i>cj0268c</i> knockout mutant (NCTC 11168::<i>cj0268c</i>), (3) complemented knockout mutant (NCTC 11168::<i>cj0268c-</i>comp-<i>cj0268c</i>). The numbers of colonies of the respective fourth dilution plated onto blood agar are shown. The <i>cj0268c</i> knockout mutant shows a clearly diminished resistance to Triton X-100. When defining the number of wild type colonies as 100%, the mean value of colonies recovered from the corresponding <i>cj0268c</i>-mutant was 21.3% (±9.02, P<0.0039). After complementation of the mutant with an intact copy of <i>cj0268c</i> the percentage of colonies obtained increased to 91.3%.</p

Additional file 4: of SMRT sequencing of the Campylobacter coli BfR-CA-9557 genome sequence reveals unique methylation motifs

Prodigal annotation of the C. coli BFR-CA-9557. Annotation of the C. coli BFR-CA-9557 genome using the Prodigal/Prokka annotation pipeline. (XLS 727 kb

Additional file 3: of SMRT sequencing of the Campylobacter coli BfR-CA-9557 genome sequence reveals unique methylation motifs

RAST annotation of the C. coli BFR-CA-9557. Annotation of the C. coli BFR-CA-9557 genome using the rapid annotation using subsystem technology platform (RAST, http://rast.nmpdr.org ). (XLS 3605 kb

Le Miroir des sports : publication hebdomadaire illustrée

17 août 19371937/08/17 (N964)-1937/08/17