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Pathogen interactions with cultured fish populations are well studied, but their effects on native fishes have not been characterized. In Chile, the disease caused by bacterial species Piscirickettsia salmonis represents one of the main issues and is considered to be one of the important pathogens in the field of aquaculture. They have been found to infect native fish. Therefore, it is necessary to understand the impact of P. salmonis on native species of local commercial value, as well as the potential impact associated with the emergence of antibiotic-resistant strains of P. salmonis. Due to this purpose, the native fish Eleginops maclovinus was used in our study. Fish were randomly distributed in tanks and intraperitoneally inoculated with two strains of P. salmonis. No mortality was recorded during the experiment. Cortisol, glucose and total α-amino acid levels increased in fish injected with AUSTRAL-005 strain compared to sham-injected and LF-89-inoculated fish. Moreover, results showed an increase in the activity of carbohydrates and lipids metabolism in liver; and an increase in the carbohydrates, lipids and total α-amino acid metabolism in muscle after injection with AUSTRAL-005. Our results suggest that P. salmonis modulates the physiology of E. maclovinus and the physiological impact increase in the presence of the antibiotic-resistant strain AUSTRAL-005.

Francisella noatunensis subsp. noatunensis, the etiological agent of Francisellosis, affects a large number of farmed species such as Salmo salar. This species coexists with several native species in the same ecosystem, including Eleginops maclovinus. Our objective was to evaluate the susceptibility, presence of clinical symptoms, and the ability of Eleginops maclovinus to respond to Francisella infection. For this, healthy individuals were inoculated with 1.5 × 101, 1.5 × 105, and 1.5 × 1010 bact/μL of Francisella by intraperitoneal injection, subsequently the fish were sampled on days 1, 3, 7, 14, 21, and 28 post injection (dpi). At the end of the experiment, no mortality, nor internal and external clinical signs were observed, although in the high dose anaemia was detected. Additionally, bacteria were detected in all three doses, however there was replication at day 28 only in the liver in the high dose. Analysis of gene expression by qPCR showed that the spleen generated an immune response against infection from day 1 dpi, however at day 7 dpi most of the genes suffered repressed expression; observing over expression of the genes C3, NLRC3, NLRC5, MHCI, IgM. In contrast, expression in the anterior kidney did not vary significantly during the challenge. IgM quantification showed the production of antibodies in the medium and high doses. This study provides new knowledge about Francisella infection and the long-lasting and specific immune response generated by Eleginops maclovinus. It also demonstrates its susceptibility to Francisellosis where there is a difference in the immune response according to the tissue.

Eleginops maclovinus is a eurythermic fish that under natural conditions lives in environments with temperatures ranging from 4 to 18 °C and can be usually captured near salmon farming areas. The aim of this study was to evaluate the effect of temperature over the innate and adaptive immune response of E. maclovinus challenged with Piscirickettsia salmonis following different treatments: C (control injection with culture medium at 12 °C), C+ (bacterial injection at 12 °C), 18 °C c/A + B (injection with culture medium in acclimation at 18 °C), 18 °C c/A + B (bacterial injection in acclimation at 18 °C), 18 °C s/A + M (injection with culture medium without acclimation at 18 °C) and 18 °C s/A + B (bacterial injection without acclimation at 18 °C). Each injection had 100 μL of culture medium or with 100 μL at a concentration 1 × 108 of live bacteria, sampling six fish per group at 4, 8, 12, 16 and 20 days post-injection (dpi). Expression of the mRNA related with the innate immune response gene (TLR1, TLR5, TLR8, NLRC3, NLRC5, MyD88 and IL-1β) as well as the adaptive immune response gene (MHCI, MHCII, IgMs and IgD) were measured in spleen and head kidney. Gene expression profiles were treatment-type and time dependent. Levels of Immunoglobulin M (IgM) increased in challenged groups with P. salmonis from day 8–20 post challenge, which suggest activation of B cells IgM + through P. salmonis epitope detection. Additionally, a rise in temperature from 12 °C (C+) to 18 °C (with/without acclimation) also resulted in antibody increment detected in serum with significant differences between “18 °C c/A + B” and “18 °C s/A + B” groups. This is the first study that evaluates the effect of temperature changes and mRNA expression related with immune system gene over time on E. maclovinus, a native wild life fish that cohabits in the salmon farming environment.

Smoltification under intensive aquaculture conditions involves important physiological changes to intestinal structures that facilitate acclimation to hyperosmotic environments. This study evaluated if genic expression of primary contributors during the smoltification process in the intestine are in synchrony with gill Na+/K+-ATPase activity. Discordance was found between gill Na+/K+-ATPase activity levels and intestinal genic expressions. In particular, the intestine showed increased mRNA expression for prolactin, a hormone that participates in hypoosmotic environments, and in mRNA expression of the growth hormone, which mediates the hydromineral balance in hyperosmotic environments. These findings indicate a lack of sufficient intestinal maturation in the fish at the moment when they were transferred to seawater environments in the normal aquaculture practices. This intestinal immaturity is a possible explanation for the high rates of desadapted individuals in the seawater phase in Chilean salmon aquaculture.

Although Caligus rogercresseyi negatively impacts Chilean salmon farming, the metabolic effects of infection by this sea louse have never been completely characterized. Therefore, this study analyzed lactate responses in the plasma, as well as the liver/muscle lactate dehydrogenase (LDH) activity and gene expression, in Salmo salar and Oncorhynchus kisutch infested by C. rogercresseyi. The lactate responses of Atlantic and Coho salmon were modified by the ectoparasite. Both salmon species showed increasing in plasma levels, whereas enzymatic activity increased in the muscle but decreased in the liver. Gene expression was overexpressed in both Coho salmon tissues but only in the liver for Atlantic salmon. These results suggest that salmonids need more energy to adapt to infection, resulting in increased gene expression, plasma levels, and enzyme activity in the muscles. The responses differed between both salmon species and over the course of infection, suggesting potential species-specific responses to sea-lice infection.

Fluctuations in ambient temperature along with the presence of pathogenic microorganisms can induce important cellular changes that alter the homeostasis of ectothermic fish. The aim of this study was to evaluate how sudden or gradual changes in environmental temperature together with the administration of Piscirickettsia salmonis modulate the transcription of genes involved in cellular stress response in the liver of Eleginops maclovinus. Fish were subjected to the following experimental conditions in duplicate: C− 12 °C: Injection only with culture medium, C+ 12 °C: Injection with P. salmonis, AM 18 °C: Injection only with culture medium under acclimation at 18 °C, AB 18 °C: Injection with P. salmonis under acclimation at 18 °C, SM 18 °C: Injection only with culture medium and thermal shock at 18 °C and SB 18 °C: Injection with P. salmonis and thermal shock at 18 °C and sampling at 4-, 8-, 12-, 16- and 20-day post injection (dpi). The genes implied in the heat shock response (HSP70, HSC70, HSP90, and GRP78), apoptosis pathway (BAX and SMAC/Diablo), ubiquitination (E2, E3, ubiquitin, and CHIP), and 26 proteasome complex (PSMB7, PSMC1, and PSMA2) showed expression profiles dependent on time and type of injection applied. All the genes greatly increased their expression levels at day 16 and showed moderate increases at day 20, except for PSMA2 which showed a higher increase between 4- and 12-day post challenges. Our results suggest that the changes observed at the final days of the experiment are due to temperature more than P. salmonis.

The nucleotide binding oligomerization domain like receptors, or NOD like receptors (NLRs), are intracellular receptors responsible for recognizing pathogens in vertebrates. Several NLR mammalian models have been characterized and analyzed but few studies have been performed with teleost species. In this study, we analyzed the nucleotide sequence of six mRNA variants of NLRC3 in Atlantic salmon (SsNLRC3), and we deduced the amino acid sequence coding for two different isoforms with a total length of 1135 amino acids and 1093 amino acids. We analyzed the phylogeny of all variants, including a Piscirickettsia salmonis infection in Atlantic salmon. All variants and their expression pattern during infection were analyzed using real-time qPCR. One of the analyzed variants was over-expressed during the early stages of Piscirickettsia salmonis infection, and we were able to identify two different SsNLRC3 isoforms. Lastly, we observed that an alteration in the amino acid sequence of one of the isoforms can directly affect the pathogen recognition function.

Caligus rogercresseyi, a marine ectoparasite, causes notable economic losses for the Chilean salmonid industry. Nevertheless, the immunological responses of infected fish remain poorly understood, including proinflammatory cytokine generation and the respective modulatory effects of various cytokine receptors. This study evaluated mRNA expression of the NLRC5, major histocompatibility complex (MHC) class II, I-kappa-B-alpha, a regulatory that inhibits NF-kappa-B, and proinflammatory cytokines (IL-1&beta, and IL-18) in the liver and muscle of Atlantic salmon (Salmo salar) and Coho salmon (Oncorhynchus kisutch) during a time-course C. rogercresseyi infestation trial. All assessed mRNA were strongly regulated during infestation, but S. salar showed up-regulated expression, possibly accounting for the high infestation vulnerability of this salmonid. In conclusion, this work helps to understand the modulation of the expression of different transcripts involved over short periods of C. rogercresseyi infestation in two salmonid species (S. salar and O. kisutch).

Ferritin is a major iron storage protein essential not only in the infectious process, but also in any circumstance generating oxidative stress. In this study, the cDNA coding sequence of ferritin-H was obtained from the sub-Antarctic Notothenioid fish Eleginops maclovinus through transcriptomic analysis of the head kidney. This sequence contained a 534 bp open reading frame that coded for a 177 amino acid protein with a molecular weight of 20,786.2 Da and a theoretical pI of 5.56. The protein displayed a region of iron putative response elements in the 5′UTR, two putative ferritin iron-binding region signatures, and seven characteristic amino acids with ferroxidase functions. Phylogenetic analysis related this sequence to ferritin-H sequences of other Antarctic Notothenioid fish, sharing 96.61% similarity. Constitutive gene expression analysis in different organs revealed increased ferritin-H gene expression in the gills, spleen, muscle, and liver. After infection with two bacterial strains of Piscirickettsia salmonis (LF-89 and Austral-005), ferritin-H was differentially expressed depending on bacterial strain and tissue. This study provides relevant information towards understanding the iron metabolism of a sub-Antarctic Notothenioid fish.

The NOD-like receptors (NLRs) were recently identified as an intracellular pathogen recognition receptor family in vertebrates. While the immune system participation of NLRs has been characterized and analyzed in various mammalian models, few studies have considered NLRs in teleost species. Therefore, this study analyzed the Atlantic salmon (Salmo salar) NLRC5. Structurally, Atlantic salmon NLRC5 presented leucine-rich repeat subfamily genes. Phylogenetically, NLRC5 was moderately conserved between S. salar and other species. Real-time quantitative PCR revealed NLRC5 expression in almost all analyzed organs, with greatest expressions in the head kidney, spleen, and hindgut. Furthermore, NLRC5 gene expression decreased during smolt stage. These data suggest that NLRC5 participates in the Atlantic salmon immune response and is regulated, at least partly, by the smoltification process, suggesting that there is a depression of immune system from parr at smolt stage. This is the first report on the NLRC5 gene in salmonid smolts.

Caligus rogercresseyi sea lice negatively impact Chilean salmonid farming, but no complete characterization for the metabolic effects of Caligus infestation currently exists. Therefore, the aim of this study was to analyze the effects of C. rogercresseyi infestation on the metabolic responses of Salmo salar and Oncorhynchus kisutch. Energy metabolism responses to C. rogercresseyi were examined over a time-course infestation of both salmonid species. Plasma metabolite levels and enzymatic activities related to intermediate metabolisms of carbohydrate, amino acid, and lipid were evaluated in the liver and muscle. Plasma glucose levels changed in both salmonid species, increasing at 1–3 days post-infestation. In turn, triglyceride levels increased at days 3 and 7 in O. kisutch and S. salar, respectively, while protein and total α-amino acids increased in O. kisutch but decreased in S. salar during infestation. Amino acid intermediate catabolic metabolism in the liver and muscle of O. kisutch increased during infestation, indicating a higher use of the gluconeogenic pathway than S. salar. Lipid intermediate anabolic metabolism increased in O. kisutch liver, remained unchanged in S. salar liver, and increased 1 day post-infestation in the muscle of both salmonids. Liver and muscle carbohydrate intermediate anabolic metabolism in O. kisutch increased during infestation, suggesting that this species preferentially uses the glycogenolytic-glucolytic pathway, in contrast with S. salar. In conclusion, amino acid and carbohydrate catabolism enzymes in O. kisutch activated soon after initial sea lice infestation, which would allow this species to dispose of energy substrates earlier than S. salar. This physiological data contributes towards the ability of O. kisutch to more adeptly cope with the increased energy demand imposed by C. rogercresseyi infestation.

This study describes de novo transcriptome sequencing and annotation analyses for the head kidney of the sub-Antarctic notothenioid fish Eleginops maclovinus, a sister group of the Antarctic notothenioid fish clade. Moreover, E. maclovinus is one of the most eurythermal and euryhaline representatives of the Notothenioidei suborder. RNA-seq data were generated by the 454 GS Junior system, resulting in 11,207 contigs that were then assembled by the Genomic Workbench CLC software. The transcriptome was annotated by BLASTing each sequence against the universal, non-redundant NCBI database (National Center for Biotechnology Information) using the AUSTRAL-omics computer cluster. A significant number of transcripts related to innate and adaptive immunity were found in the sequences, which could be used as references in future immunological studies in E. maclovinus.