PMC

Specificity of the C. ruminantium pCS20 PCR assay against E. chaffeensis. Agarose gel electrophoresis of products from PCRs with C. ruminantium primers and C. ruminantium DNA (lanes 2 and 3) or E. chaffeensis DNA (lanes 4 and 5), with E. chaffeensis primers and C. ruminantium DNA (lanes 8 and 9) or E. chaffeensis DNA (lanes 10 and 11), and with general Ehrlichia primers and C. ruminantium DNA (lanes 14 and 15) or E. chaffeensis DNA (lanes 16 and 17). The 123-bp DNA standard ladder and negative-control reactions for the C. ruminantium, E. chaffeensis, and general Ehrlichia primers are shown in lanes L, 1, 7, and 13, respectively. Lanes 6 and 12 are empty.

Infection of ISE6 cells with B. burgdorferi s.s. and E. ruminantium. (A) B. burgdorferi spirochaete (arrow) “interacting” with an ISE6 cell containing a colony of E. ruminantium (arrowhead). (B) Quantification of E. ruminantium in ISE6 cells infected with B. burgdorferi followed 24 h later by E. ruminantium. Shaded bars: cells infected with B. burgdorferi and E. ruminantium; white bars: cells infected with E. ruminantium alone. The amount of E. ruminantium DNA detected by real-time PCR was significantly higher in cells with B. burgdorferi than in cells with E. ruminantium alone at 24 and 72 h p.i. (Student's t-test) and across all time points (ANOVA, p = 0.0012). Experiment was repeated twice with similar results; 3 replicates per treatment, error bars represent standard deviation. * indicates p value <0.05 by Student's t-test.

Proteins differentially expressed by E. ruminantium in BAE cells from 24 to 120 hpi based on 1DE-nLC-MS/MS. (A) Number of E. ruminantium proteins detected at each time point post-infection. (B) Hierarchical cluster analysis of differentially regulated E. ruminantium proteins at each time point post-infection, and (C) relative abundance of E. ruminantium proteins detected from 24 to 120 hpi.

ACT alignment of the E. ruminantium Kümm2 sequence (bottom sequence in each panel) against E. ruminantium Welgevonden (top sequence in each panel) to depict the expansion of membrane family 3 in the Kümm2 isolate. The coloured lines drawn between the sequences show the location of homologous genes in the same (red) or opposite (blue) orientation. Top (a) Four E. ruminantium Welgevonden open reading frames matching open reading frames in two locations (yellow lines) in the E. ruminantium Kümm2 annotation. Middle (b) The syntenic matches of the four Welgevonden ORFs to seven Kümm2 open reading frames. Bottom (c) The same four Welgevonden open reading frames matching five Kümm2 open reading frames upstream.