Antibacterial activity of asCATH1 and 2 against three species of bacteria - <i>Y. ruckeri</i> (A), <i>V. anguillarum</i> (B) and <i>E. coli</i> (C).

<p>Bacteria were grown to mid-log phase then incubated for 18 h with varying concentrations of the Atlantic salmon cathelicidins. LL-37 was included as a control for bacterial killing and <i>E. coli</i> ATCC 25922 was included as a control bacterium. Absorbance was read at 600 nm and percentage growth was calculated by comparison to the no inhibition control lane in the assay. The MIC was defined as 50% inhibition, which is indicated by a dotted line on each plot. Data shown are the mean ± SE of triplicate wells.</p

Kinetic profile of IL-8, IL-1 and IL-18 gene expression in Atlantic salmon peripheral blood leukocytes stimulated with asCATH1 (A,C,E) and asCATH2 (B,D,E).

<p>Gene expression was assessed by real-time PCR at 0, 2, 5, 8 and 16 h post incubation with asCATH1 or asCATH2 at 2.5 µM (A,B), 5 µM (C,D) and 10 µM (E,F). Expression was normalised to the mean expression of four stable reference genes. Data shown are the means ± SE of quadruplicate PBL samples assayed in duplicate by quantitative real-time PCR and presented as fold induction compared to 0 h. * indicates a significant fold induction (P<0.05) compared to 0 h while the dotted line represents the minimum fold change (2 fold) deemed biologically significant.</p

Haemolytic activity of the Atlantic salmon cathelicidins to Atlantic salmon erythrocytes.

<p>Erythrocytes were incubated with varying concentrations of the cathelicidins for 2 h and the absorbance of the well supernatants was read at 405 nm to detect released haemoglobin. Percentage haemolysis was calculated from 100% lysis controls, which were incubated with 0.2% Triton-X 100. Data shown are the means ± SD of duplicate wells.</p

Expression of asCATH1 and 2 in a range of organs from a representative healthy juvenile Atlantic salmon.

<p>PCR was performed on cDNA samples from the various organs and 10 µl of each product was visualised on a 1% LB agarose gel. β-actin was included to assess the quality and quantity of cDNA.</p

Antibacterial activity of asCATH1 and 2.

<p>Bacteria were incubated with varying concentrations of cathelicidin peptides for 18 h before absorbance was read at 600 nm to determine bacterial growth. MIC values are shown as the range of concentrations which produced a 50% reduction in growth compared to a control without cathelicidin.</p

Oligonucleotide primers used in real-time pcr experiments.

<p>Oligonucleotide primers used in real-time pcr experiments.</p

Relative expression of asCATH1 and 2 in the gill (A) and spleen (B) of Atlantic salmon post-infection with <i>Y. ruckeri</i>.

<p>Gene expression was measured by quantitative real-time PCR. Expression at 8, 24, 48, 72 and 96 h post-infection was compared to expression at 0 h. Each bar represents the mean ± SE of at least 4 fish sampled at each time point. * indicates a significant upregulation of cathelicidin expression compared to expression at 0 h (p<0.05, as assessed by one-way analysis of variance and Dunnett's post-test).</p

The chemokine receptor CXCR4 promotes granuloma formation by sustaining a mycobacteria-induced angiogenesis programme

CXC chemokine receptor 4 plays a critical role in chemotaxis and leukocyte differentiation. Furthermore, there is increasing evidence that links this receptor to angiogenesis. Using the well-established zebrafish-Mycobacterium marinum model for tuberculosis, angiogenesis was recently found to be important for the development of cellular aggregates called granulomas that contain the mycobacteria and are the hallmark of tuberculosis disease. Here, we found that initiation of the granuloma-associated proangiogenic programme requires CXCR4 signalling. The nascent granulomas in cxcr4b-deficient zebrafish embryos were poorly vascularised, which in turn also delayed bacterial growth. Suppressed infection expansion in cxcr4b mutants could not be attributed to an overall deficient recruitment of leukocytes or to different intramacrophage bacterial growth rate, as cxcr4b mutants displayed similar microbicidal capabilities against initial mycobacterial infection and the cellular composition of granulomatous lesions was similar to wildtype siblings. Expression of vegfaa was upregulated to a similar extent in cxcr4b mutants and wildtypes, suggesting that the granuloma vascularisation phenotype of cxcr4b mutants is independent of vascular endothelial growth factor

Additional file 8: of De novo assembly of Sockeye salmon kidney transcriptomes reveal a limited early response to piscine reovirus with or without infectious hematopoietic necrosis virus superinfection

Putative DEGs identified by preliminary DESeq and edgeR analysis and subsequent qPCR validation. (PDF 1454 kb

Additional file 6: of De novo assembly of Sockeye salmon kidney transcriptomes reveal a limited early response to piscine reovirus with or without infectious hematopoietic necrosis virus superinfection

Associated UniProt 2013_11 protein database annotation of all Sockey salmon head kidney unigene components. (RAR 14953 kb