Copper induces an oxidative stress condition in the marine alga Ulva compressa that is due to the production of superoxide anions and hydrogen peroxide, mainly in organelles. The increase in hydrogen peroxide is accompanied by increases in intracellular calcium and nitric oxide, and there is a crosstalk among these signals. The increase in intracellular calcium activates signaling pathways involving Calmodulin-dependent Protein Kinases (CaMKs) and Calcium-Dependent Protein Kinases (CDPKs), leading to activation of gene expression of antioxidant enzymes and enzymes involved in ascorbate (ASC) and glutathione (GSH) synthesis. It was recently shown that copper also activates Mitogen-Activated Protein Kinases (MAPKs) that participate in the increase in the expression of antioxidant enzymes. The increase in gene expression leads to enhanced activities of antioxidant enzymes and to enhanced levels of ASC and GSH. In addition, copper induces an increase in photosynthesis leading to an increase in the leve of Nicotinamide Adenine Dinucleotide Phosphate (NADPH). Copper also induces an increase in activities of enzymes involved in C, N, and S assimilation, allowing the replacement of proteins damaged by oxidative stress. The accumulation of copper in acute exposure involved increases in GSH, phytochelatins (PCs), and metallothioneins (MTs) whereas the accumulation of copper in chronic exposure involved only MTs. Acute and chronic copper exposure induced the accumulation of copper-containing particles in chloroplasts. On the other hand, copper is extruded from the alga with an equimolar amount of GSH. Thus, the increases in activities of antioxidant enzymes, in ASC, GSH, and NADPH levels, and in C, N, and S assimilation, the accumulation of copper-containing particles in chloroplasts, and the extrusion of copper ions from the alga constitute essential mechanisms that participate in the buffering of copper-induced oxidative stress in U. compressa.

The genome of the marine alga Ulva compressa was assembled using long and short reads. The genome assembly was 80.8 Mb in size and encoded 19,207 protein-coding genes. Several genes encoding antioxidant enzymes and a few genes encoding enzymes that synthesize ascorbate and glutathione were identified, showing similarity to plant and bacterial enzymes. Additionally, several genes encoding signal transduction protein kinases, such as MAPKs, CDPKS, CBLPKs, and CaMKs, were also detected, showing similarity to plants, green microalgae, and bacterial proteins. Regulatory transcription factors, such as ethylene- and ABA-responsive factors, MYB, WRKY, and HSTF, were also present and showed similarity to plant and green microalgae transcription factors. Genes encoding enzymes that synthesize ACC and ABA-aldehyde were also identified, but oxidases that synthesize ethylene and ABA, as well as enzymes that synthesize other plant hormones, were absent. Interestingly, genes involved in plant cell wall synthesis and proteins related to animal extracellular matrix were also detected. Genes encoding cyclins and CDKs were also found, and CDKs showed similarity to animal and fungal CDKs. Few genes encoding voltage-dependent calcium channels and ionotropic glutamate receptors were identified as showing similarity to animal channels. Genes encoding Transient Receptor Potential (TRP) channels were not identified, even though TRPs have been experimentally detected, indicating that the genome is not yet complete. Thus, protein-coding genes present in the genome of U. compressa showed similarity to plant and green microalgae, but also to animal, bacterial, and fungal genes.

Filifolinone is an aromatic geranyl derivative, a natural compound isolated from Heliotropum sclerocarpum, which has immunomodulatory effects on Atlantic salmon, upregulating cytokines involved in Th1-type responses through a mechanism that remains unknown. In this work, we determined whether the immunomodulatory effects of filifolinone depend on the host microbiotic composition. We evaluated the effect of filifolinone on immune genes and intestinal microbiotic composition of normal fish and fish previously treated with bacitracin/neomycin. Filifolinone induced the early expression of IFN-α1 and TGF-β, followed by the induction of TNF-α, IL-1β, and IFN-γ. A pre-treatment with antibiotics modified this effect, mainly changing the expression of IL-1β and IFN-γ. The evaluation of microbial diversity shows that filifolinone modifies the composition of intestinal microbiota, increasing the abundance of immunostimulating organisms like yeast and firmicutes. We identified 69 operational taxonomic units (OTUs) associated with filifolinone-induced IFN-γ. Our results indicate that filifolinone stimulates the immune system in two ways, one dependent on fish microbiota and the other not. To our knowledge, this is the first report of microbiota-dependent immunostimulation in Salmonids.

In order to analyze the mechanisms involved in copper accumulation in Ulva compressa, algae were collected at control sites of central and northern Chile, and at two copper-polluted sites of northern Chile. The level of intracellular copper, reduced glutathione (GSH), phytochelatins (PCs), PC2 and PC4, and transcripts encoding metallothioneins (MTs) of U. compressa, UcMT1, UcMT2 and UcMT3, were determined. Algae of control sites contained around 20 $\mathsf{\mu }$g of copper g⁻¹ of dry tissue (DT) whereas algae of copper-polluted sites contained 260 and 272 $\mathsf{\mu }$g of copper g⁻¹ of DT. Algae of control sites and copper-polluted sites did not show detectable amounts of GSH, the level of PC2 did not change among sites whereas PC4 was increased in one of the copper-polluted sites. The level of transcripts of UcMT1 and UcMT2 were increased in algae of copper-polluted sites, but the level of UcMT3 did not change. Algae of a control site and a copper-polluted site were visualized by transmission electron microscopy (TEM) and the existence of copper in electrodense particles was analyzed using energy dispersive x-ray spectroscopy (EDXS). Algae of copper-polluted sites showed electrodense nanoparticles containing copper in the chloroplasts, whereas algae of control sites did not. Algae of a control site, Cachagua, were cultivated without copper (control) and with 10 $\mathsf{\mu }$M copper for 5 days and they were analyzed by TEM-EDXS. Algae cultivated with copper showed copper-containing nanoparticles in the chloroplast whereas control algae did not. Thus, U. compressa from copper-polluted sites exhibits intracellular copper accumulation, an increase in the level of PC4 and expression of UcMTs, and the accumulation of copper-containing particles in chloroplasts.

Andrographolide, fucoidan, or a combination of both compounds were evaluated to determine their effects on the antiviral response in the Atlantic salmon macrophage-like cell line (SHK-1) infected with infectious pancreatic necrosis virus (IPNV). We assessed the transcript expression levels of key molecules involved in the interferon (IFN)-dependent antiviral response, as well as the viral load in cells treated with these compounds. In non-infected cells, incubation with either fucoidan, andrographolide, or a mixture of both resulted in an increase in the transcript expression of IFNα1 and various interferon-stimulated genes (ISGs). In IPNV-infected cells, treatment with either fucoidan or andrographolide separately did not significantly enhance the antiviral response compared to that of infected cells that had not previously been treated with these compounds. In contrast, the combination of andrographolide and fucoidan led to a marked increase in the transcript expression of viperin and a significant reduction in viral load. Overall, combining andrographolide and fucoidan resulted in a greater reduction in IPNV viral load in infected cells than that noted when the compounds were administered individually. Our findings suggest that pre-incubation with this mixture promotes the establishment of a protective antiviral state against IPNV, likely mediated by an IFN-dependent response.

Infectious pancreatic necrosis virus (IPNV), a member of the family Birnaviridae, is a major pathogen of farmed salmonids and an important model in fish virology. Despite its small genome, which encodes only five viral proteins, IPNV exhibits complex molecular processes that govern genome expression, replication, and particle assembly. Comprehensive descriptions of the molecular biology and replication cycle of IPNV were largely established in reviews published in the mid-1990s, whereas more recent reviews have primarily focused on virulence determinants, epidemiology, or host–virus interactions. This review provides an updated synthesis of available experimental knowledge on the molecular biology of IPNV by integrating classical and recent studies addressing virion architecture, genome organization, and the functions of viral proteins. Particular attention is given to the molecular events involved in the viral replication cycle, including virus entry, genome transcription, translation and replication in the cytoplasm, polyprotein processing by the viral protease, and the coordination between genome replication and virion assembly. When appropriate, experimental observations from the related Avibirnavirus infectious bursal disease virus are considered to provide additional context for molecular mechanisms conserved within the family Birnaviridae. Together, these studies outline the current understanding of the molecular processes governing IPNV replication and morphogenesis.