Fig 1.
Schematic representation of the experimental design showing E. ruminantium acquisition and transmission by A. hebraeum ticks as well as the downstream analysis of the resultant tick midgut transcriptomes by RNA-seq analysis.
The figure was created and exported under a paid subscription with BioRender.com.
Fig 2.
Monitoring of rectal temperatures in experimental and control sheep during E. ruminantium acquisition and transmission by A. hebraeum ticks.
Panel A shows the monitored rectal temperatures of sheep during the acquisition of E. ruminantium by A. hebraeum nymphs. The test sheep (depicted in red) was administered an intravenous inoculum of E. ruminantium at day 0, while the control sheep (depicted in blue) did not receive an inoculum. Two batches of A. hebraeum nymphs were allowed to feed on both sheep on days 2 and 4 post-inoculation. The experiment concluded at day 15, following the nymphs’ completion of feeding. Panel B displays the rectal temperatures of a new set of naïve sheep during E. ruminantium transmission by the emerging A. hebraeum adults. Adults potentially infected with E. ruminantium were allowed to feed on test-sheep 2 to mimic a natural transmission, while those from the control-sheep 1 fed on control-sheep 2 (shown in blue).
Table 1.
Mapping statistics of E. ruminantium reads in infected midgut groups of A. hebraeum ticks.
Table 2.
Statistical summary of Amblyomma hebraeum midgut sequencing of ten RNA samples in five comparison groups.
Fig 3.
Species classification of BLASTx matches: A pie chart representation of A. hebraeum transcriptome unigenes.
Fig 4.
Differential gene expression analysis in E. ruminantium infected A. hebraeum midguts visualized as a volcano plot.
This plot compares the DEGs across five E. ruminantium infected A. hebraeum midgut groups to respective controls. Red dots signify significantly up-regulated genes, blue dots represent significantly down-regulated genes, and blank points denote genes with no significant change in expression. Significance was determined based on a log2FC ≥ 1.5 or log2FC ≤ -1.5 and a q-value < 0.05. The y-axis plots the logCPM values, providing a measure of each gene’s expression level (Counts Per Million).
Table 3.
Selected top DEGs up-regulated in E. ruminantium infected A. hebraeum nymph midgut tissues at 14 days post detachment from host.
Table 4.
Selected top DEGs up-regulated in E. ruminantium infected A. hebraeum unfed male midgut tissues.
Table 5.
Selected top DEGs up-regulated in E. ruminantium infected A. hebraeum unfed female midgut tissues.
Fig 5.
UpsetR plot of differential gene expression in E. ruminantium infected A. hebraeum midgut tissue.
This figure illustrates the number of up-regulated and down-regulated DEGs common to the five midgut comparison groups. It highlights the unique and shared DEGs in nymphs, unfed and partly-fed male and female tick midguts.
Table 6.
Selected top DEGs up-regulated in E. ruminantium infected A. hebraeum partly-fed male midgut tissues.
Table 7.
Selected top DEGs up-regulated in E. ruminantium infected A. hebraeum partly-fed female midgut tissues.
Fig 6.
KEGG pathway analysis of selected categories of A. hebraeum midgut transcriptional response to E. ruminantium infection.
Panel [A] depicts the comparative analysis of DEGs in the midguts of infected nymphs against uninfected controls. Panel [B] elucidates the DEG patterns in infected partly-fed females compared to their respective controls and [C] infected unfed females compared to control.
Fig 7.
Selected regression plots comparing the differential expression values of eight genes in A. hebraeum nymphs, and the unfed male and female tick midguts.
This comparison is between the quantitative real-time PCR (qPCR) data and the RNA-seq data. The log2 fold-change values from qPCR are plotted on the x-axis and the corresponding values from RNA-Seq are plotted on the y-axis. The plot illustrates the correlation and consistency between the qPCR validation and the original RNA-Seq data for these selected genes. All validation plots are detailed in S2 Data.
Table 8.
Selected DEGs values of 8 genes of A. hebraeum unfed male midgut. The log2 ratios qPCR plotted against those of respective RNA-Seq.