Relationship between respiratory response and metabolic pathways of S. Derby and S. Mbandaka.

<p>Respiratory responses which distinguish <i>S</i>. Derby and <i>S</i>. Mbandaka under aerobic conditions at 25°C and the corresponding metabolic pathway for the metabolism of the compound from uptake to glycolysis. Only <i>S</i>. Mbandaka respires on D-galactonic acid-g-lactone (DGL) (a), this can be attributed to the lack of pathway genes (e) (KEGG map 00052, created 31/5/12). Likewise, <i>S</i>. Derby is unable to respire on D-glucosaminic acid (DGA) while <i>S</i>. Mbandaka can (c) yet both serovars possess the same pathway genes (g) (KEGG map 00030, created 9/3/13). For both DGL and DGA there were gaps in the SEEDmodel reconstruction which prevented the metabolism of the compounds, these were filled using ECBLAST and SEEDviewer BLASTp (yellow stars). Both serovars contain the same pathway genes for the metabolism of mucic acid (f) (KEGG map 00053, created 28/9/09) yet the area under the respiratory curve for this compound, and not the log phase gradient or lag phase, was significantly greater for <i>S</i>. Derby (b).</p

Multiplex PCR results.xls

<p>276 APEC were investigated for R type, phylogroup and virulence factors using multiplex pcr as described in </p><p> </p><p>Comparative genomics of European Avian Pathogenic E. coli (APEC) (in press)</p

Expression of SPI-23 in <i>S</i>. Derby D1 when exposed to jejunum and colon explants.

<p>Fold differences in qRT-PCR expression levels +/−1SEM of ten genes found on SPI-23, in the order they appear on the island, when exposed to porcine jejunum and colon explants relative to a no tissue control.</p

Association and invasion of IPEC-J2 monolayers by <i>S</i>. Derby and <i>S</i>. Mbandaka.

<p>(<b>a</b>) Association (cells adhering and invading) and (<b>b</b>) invasion of IPEC-J2, porcine jejunum derived cell line, by <i>S</i>. Derby strains D1 and D2 and <i>S</i>. Mbandaka strains M1 and M2 after 15, 30 and 60 minute incubation periods. Values are mean colony forming units (CFU) +/−1SEM recovered from disrupted monolayers planted on to LB agar plates and incubated for 16 hours at 37°C.</p

Comparison of morphological and structural features of the <i>S</i>. Derby D1 parental and <i>potR</i> mutant strains.

<p>Comparison of <i>S</i>. Derby D1 (left) and mutant strain <i>S</i>. Derby D1 <i>ΔpotR::kan</i> (right). (<b>a</b>) Strains were photographed after 2 hours remaining static post 16 hours of culturing at 37°C with agitation at 220 rpm. (<b>b</b>) Negative electron microscopy performed after 16 hours of culturing at 37°C with agitation at 220 rpm. Pili on the mutant and type-1 fimbriae on the parental strain are marked by arrows. (<b>c</b>) Strains were plated on to LB agar plates and incubated for 16 hours at 37°C.</p

Adhesion and Invasion of <i>S</i>. Derby D1 parent and <i>potR</i> mutant strains.

<p>Differential stain confocal microscopy of (<b>a</b>) <i>S</i>. Derby D1 and (<b>b</b>) <i>S</i>. Derby D1 <i>ΔpotR::kan</i> that have adhered (yellow) and invaded (red) IPEC-J2 monolayers (blue) after 4 hours exposure.</p

Isolates were grouped into 114 clusters at the 40% similarity level.

<p>Isolates were grouped into 114 clusters at the 40% similarity level.</p

Simpson’s Index of Diversity (1-D) of isolates from different categories (country and host species) at the similarity levels from 25% to 90%.

<p>Simpson’s Index of Diversity (1-D) of isolates from different categories (country and host species) at the similarity levels from 25% to 90%.</p

Minimal spanning tree constructed based on the MLST profiles of the 313 isolates (Bionumerics V6) and coloured according to the cluster numbers: red, cluster 76; purple, cluster 103; light blue, cluster 30; dark green, cluster 42; green, no array data and white, a mixture of rare cluster number.

<p>Minimal spanning tree constructed based on the MLST profiles of the 313 isolates (Bionumerics V6) and coloured according to the cluster numbers: red, cluster 76; purple, cluster 103; light blue, cluster 30; dark green, cluster 42; green, no array data and white, a mixture of rare cluster number.</p