In vitro effects of phytogenic feed additive on Piscirickettsia salmonis growth and biofilm formation.

Piscirickettsiosis is the main cause of mortality in salmonids of commercial importance in Chile, which is caused by Piscirickettsia salmonis, a Gram-negative, γ-proteobacteria that can produce biofilm as one of its virulence factors. The Chilean salmon industry uses large amounts of antibiotics to control piscirickettsiosis outbreaks, which has raised concern about its environmental impact and the potential to induce antibiotic resistance. Thus, the use of phytogenic feed additives (PFA) with antibacterial activity emerges as an interesting alternative to antimicrobials. Our study describes the antimicrobial action of an Andrographis paniculate-extracted PFA on P. salmonis planktonic growth and biofilm formation. We observed complete inhibition of planktonic and biofilm growth with 500 and 400 µg/mL of PFA for P. salmonis LF-89 and EM-90-like strains, respectively. Furthermore, 500 µg/mL of PFA was bactericidal for both evaluated bacterial strains. Sub-inhibitory doses of PFA increase the transcript levels of stress (groEL), biofilm (pslD), and efflux pump (acrB) genes for both P. salmonis strains in planktonic and sessile conditions. In conclusion, our results demonstrate the antibacterial effect of PFA against P. salmonis in vitro, highlighting the potential of PFA as an alternative to control Piscirickettsiosis.

Hypertensive Nephropathy: Unveiling the Possible Involvement of Hemichannels and Pannexons.

Hypertension is one of the most common risk factors for developing chronic cardiovascular diseases, including hypertensive nephropathy. Within the glomerulus, hypertension causes damage and activation of mesangial cells (MCs), eliciting the production of large amounts of vasoactive and proinflammatory agents. Accordingly, the activation of AT1 receptors by the vasoactive molecule angiotensin II (AngII) contributes to the pathogenesis of renal damage, which is mediated mostly by the dysfunction of intracellular Ca2+ ([Ca2+]i) signaling. Similarly, inflammation entails complex processes, where [Ca2+]i also play crucial roles. Deregulation of this second messenger increases cell damage and promotes fibrosis, reduces renal blood flow, and impairs the glomerular filtration barrier. In vertebrates, [Ca2+]i signaling depends, in part, on the activity of two families of large-pore channels: hemichannels and pannexons. Interestingly, the opening of these channels depends on [Ca2+]i signaling. In this review, we propose that the opening of channels formed by connexins and/or pannexins mediated by AngII induces the ATP release to the extracellular media, with the subsequent activation of purinergic receptors. This process could elicit Ca2+ overload and constitute a feed-forward mechanism, leading to kidney damage.

Biofilm Produced In Vitro by Piscirickettsia salmonis Generates Differential Cytotoxicity Levels and Expression Patterns of Immune Genes in the Atlantic Salmon Cell Line SHK-1.

Piscirickettsia salmonis is the causative agent of Piscirickettsiosis, an infectious disease with a high economic impact on the Chilean salmonid aquaculture industry. This bacterium produces biofilm as a potential resistance and persistence strategy against stressful environmental stimuli. However, the in vitro culture conditions that modulate biofilm formation as well as the effect of sessile bacteria on virulence and immune gene expression in host cells have not been described for P. salmonis. Therefore, this study aimed to analyze the biofilm formation by P. salmonis isolates under several NaCl and iron concentrations and to evaluate the virulence of planktonic and sessile bacteria, together with the immune gene expression induced by these bacterial conditions in an Atlantic salmon macrophage cell line. Our results showed that NaCl and Fe significantly increased biofilm production in the LF-89 type strain and EM-90-like isolates. Additionally, the planktonic EM-90 isolate and sessile LF-89 generated the highest virulence levels, associated with differential expression of il-1ß, il-8, nf-κb, and iκb-α genes in SHK-1 cells. These results suggest that there is no single virulence pattern or gene expression profile induced by the planktonic or sessile condition of P. salmonis, which are dependent on each strain and bacterial condition used.

Salmo salar glucocorticoid receptors analyses of alternative splicing variants under stress conditions.

Cortisol is the main corticosteroid in teleosts, exerting multiple functions by activating glucocorticoid receptors (GR). Most teleost species have two GR genes, gr-1 and gr-2. Some teleost also presents two splice variants for gr-1; gr-1a and gr-1b. In this study, we report for first time the presence of 2 homeologous genes for gr-1 and gr-2, located on chromosomes 4q-13q (gr-1) and 5p-9q (gr-2) of the Salmo salar genome. Furthermore, our results describe gr-1 splice variants derived from chromosome 4 and 13, sharing typical teleost GR elements such as the 9 amino acid insertion in the DNA binding domain (DBD) and variations in the length of the ligand binding domain (LBD). Three splice variants were predicted for the gr-2 homeologous gene in chromosome 5, with differences of a 5 amino acid insertion in the DBD. We also identified an uncommon truncated gr-2 gene in chromosome 9 in salmon, which lacked the DBD and LBD domains. Finally, by designing specific primers for each predicted splice variant, we validated and evaluated the expression of their transcripts in S. salar subjected to stress caused by stocking density. Differences were observed in the expression of all identified mRNAs, revealing that gr-1 and gr-2 splice variants were upregulated in head kidney and gills of post-stressed fish. In conclusion, our findings suggest that from specific salmonid genomic duplication (125 MYA), two gene copies of each GR receptor were generated in S. salar. The identified splice variants could contribute to the variability of GR receptor complex modulation expression during stressful events, leading to variations in physiological responses in fish.