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Transgenerational immune priming (TGIP) is a phenomenon by which an initial exposure to a pathogen, here Tetracapsuloides bryosalmonae (causative agent of Proliferative Kidney Disease), stimulates the subsequent immune response to the same or a different pathogen in future generations. The impact of rearing conditions in previous generations, regardless of their exposure to the pathogen, on the immune cell composition and immune response in subsequent generations has not yet been investigated in the brown trout- T. bryosalmonae system. In the present work, we performed flow cytometry to analyze immune cell populations of brown trout. Parental generations (F0) differed in rearing conditions and exposure to the parasite. The study evaluated the baseline frequencies of IgM + B cells, myeloid cells, and CD8 + T cells in the offspring (F1) young-of-the-year brown trout. Afterwards, F1 fish were experimentally infected with T. bryosalmonae spores and monitored during eight weeks post-infection. The kidney was identified as an immune-cell niche dominated by myeloid cells, which represent approximately two-thirds of the total immune cell population, along with a substantial proportion of IgM + B cells. CD8 + T cells constitute only a minor fraction within this niche. As measured by flow cytometry, the immune-cell frequencies of offspring were largely unaffected by the parental rearing background (F0) and infection history. Parental history had no influence on the outcome of experimental infection. In T. bryosalmonae exposed animals, parasite concentration increased significantly over time. Moreover, a proportional increase in IgM + B cells and a proportional decrease in myeloid cells over time was observed. However, the increase in IgM + B cells was also detected in control animals. In conclusion, this study presents the first analysis of immune cell composition in F1 brown trout derived from parents reared under three distinct environmental conditions with varying parasite exposures. Our flow cytometry results highlight the need for alternative approaches to investigate transgenerational immune priming (TGIP) in brown trout.

In fish, the skin is a multifunctional organ and the first barrier against pathogens. Salmonids differ in their susceptibility to microorganisms due to varied skin morphology and gene expression patterns. The brown trout is a salmonid species with important commercial and ecological value in Europe. However, there is a lack of knowledge regarding the genes involved in the immune response and mucus secretion in the skin of this fish. Thus, we characterized the skin transcriptome of anadromous brown trout using next-generation sequencing (NGS). A total of 1,348,306 filtered reads were obtained and assembled into 75,970 contigs. Of these contigs 48.57% were identified using BLAST tool searches against four public databases. KEGG pathway and Gene Ontology analyses revealed that 13.40% and 34.57% of the annotated transcripts, respectively, represent a variety of biological processes and functions. Among the identified KEGG Orthology categories, the best represented were signal transduction (23.28%) and immune system (8.82%), with a variety of genes involved in immune pathways, implying the differentiation of immune responses in the trout skin. We also identified and transcriptionally characterized 8 types of mucin proteins-the main structural components of the mucosal layer. Moreover, 140 genes involved in mucin synthesis were identified, and 1,119 potential simple sequence repeats (SSRs) were detected in 3,134 transcripts.

Yersinia ruckeri is the causative agent of enteric redmouth disease, a bacterial infection of marine and freshwater fish. The disease mainly affects salmonids, and outbreaks have significant economic impact on fish farms all over the world. Vaccination routines are in place against the major serotypes of Y. ruckeri but are not effective in all cases. Despite the economic importance of enteric redmouth disease, a detailed molecular understanding of the disease is lacking. A considerable number of mostly omics-based studies have been performed in recent years to identify genes related to Y. ruckeri virulence. This review summarizes the knowledge on Y. ruckeri virulence factors. Understanding the molecular pathogenicity of Y. ruckeri will aid in developing more efficient vaccines and antimicrobial compounds directed against enteric redmouth disease.

Antimicrobial peptides (AMPs) are an important part of non-specific immunity and play a key role in the cellular host defense against pathogens and tissue injury infections. We investigated the effects of AMP supplementation on the antioxidant capacity, non-specific immunity, and gut microbiota of tsinling lenok trout. 240 fish were fed diets (CT, A120, A240 and A480) containing different amounts of AMP peptides (0, 120 mg kg −1 , 240 mg kg −1 , 480 mg kg −1 ) for 8 weeks. Our results showed that the activity of total antioxidant capacity (T-SOD) and glutathione peroxidase (GSH-Px), lysozyme (LZM), catalase (CAT) and acid phosphatase (ACP) in the A240 and A480 group were higher than that in the CT group ( P  < 0.05). The content of malondialdehyde (MDA) in AMP group was significantly lower than that in CT group ( P  < 0.05). Furthermore, we harvested the mid-gut and applied next-generation sequencing of 16S rDNA. The results showed that the abundance of Halomonas in AMP group was significantly lower than that in CT group. Functional analysis showed that the abundance of chloroalkane and chloroalkene degradation pathway increased significantly in AMP group. In conclusion, AMP enhanced the antioxidant capacity, non-specific immunity, and intestinal health of tsinling lenok trout.

Background Tetracapsuloides bryosalmonae is a myxozoan parasite which causes economically important and emerging proliferative kidney disease (PKD) in salmonids. Brown trout, Salmo trutta is a native fish species of Europe, which acts as asymptomatic carriers for T. bryosalmonae . There is only limited information on the molecular mechanism involved in the kidney of brown trout during T. bryosalmonae development. We employed RNA sequencing (RNA-seq) to investigate the global transcriptome changes in the posterior kidney of brown trout during T. bryosalmonae development. Methods Brown trout were exposed to the spores of T. bryosalmonae and posterior kidneys were collected from both exposed and unexposed control fish. cDNA libraries were prepared from the posterior kidney and sequenced. Bioinformatics analysis was performed using standard pipeline of quality control, reference mapping, differential expression analysis, gene ontology, and pathway analysis. Quantitative real time PCR was performed to validate the transcriptional regulation of differentially expressed genes, and their correlation with RNA-seq data was statistically analyzed. Results Transcriptome analysis identified 1169 differentially expressed genes in the posterior kidney of brown trout, out of which 864 genes (74%) were upregulated and 305 genes (26%) were downregulated. The upregulated genes were associated with the regulation of immune system process, vesicle-mediated transport, leucocyte activation, and transport, whereas the downregulated genes were associated with endopeptidase regulatory activity, phosphatidylcholine biosynthetic process, connective tissue development, and collagen catabolic process. Conclusion To our knowledge, this is the first RNA-seq based transcriptome study performed in the posterior kidney of brown trout during active T. bryosalmonae development. Most of the upregulated genes were associated with the immune system process, whereas the downregulated genes were associated with other metabolic functions. The findings of this study provide insights on the immune responses mounted by the brown trout on the developing parasite, and the host molecular machineries modulated by the parasite for its successful multiplication and release.

With respect to salmonid aquaculture, one of the most important bacterial pathogens due to high mortality and antibiotic usage is the causative agent of typical furunculosis, spp. ( ). In Atlantic salmon, , the host response during infections with is well-documented, with furunculosis outbreaks resulting in significant mortality in commercial settings. However, less is known about the host-pathogen interactions in the emerging aquaculture species, Arctic charr . Furthermore, there is no data on the efficacy or response of this species after vaccination with commonly administered vaccines against furunculosis. To this end, we examined the immunological response of during infection with , with or without administration of vaccines (Forte Micro®, Forte Micro® + Renogen®, Elanco Animal Health). Artic charr (vaccinated or unvaccinated) were i.p.-injected with a virulent strain of (10 CFUs/mL) and tissues were collected pre-infection/post-vaccination, 8, and 29 days post-infection. Unvaccinated Arctic charr were susceptible to with 72% mortalities observed after 31 days. However, there was 72-82% protection in fish vaccinated with either the single or dual-vaccine, respectively. Protection in vaccinated fish was concordant with significantly higher serum IgM concentrations, and following RNA sequencing and transcriptome assembly, differential expression analysis revealed several patterns and pathways associated with the improved survival of vaccinated fish. Most striking was the dramatically higher basal expression of complement/coagulation factors, acute phase-proteins, and iron hemostasis proteins in pre-challenged, vaccinated fish. Remarkably, following infection, this response was abrogated and instead the transcriptome was characterized by a lack of immune-stimulation compared to that of unvaccinated fish. Furthermore, where pathways of actin assembly and FcγR-mediated phagocytosis were significantly differentially regulated in unvaccinated fish, vaccinated fish showed either the opposite regulation (ForteMicro®), or no impact at all (ForteMicro®Renogen®). The present data indicates that vaccine-induced protection against relies on the pre-activation and immediate control of humoral immune parameters that is coincident with reduced activation of apoptotic (e.g., NF-κB) and actin-associated pathways.

We studied the effect of several lactic acid bacteria (LAB) on the humoral response of brown trout (Salmo trutta). LAB groups (Lactococcus (Lc.) lactis ssp. lactis, Lactobacillus (Lb.) sakei and Leuconostoc (Leu.) mesenteroides) were administered orally at 106 colony-forming units/g feed to brown trout for 2 weeks, after which fish were switched to an unsupplemented feed. Blood and intestinal samples were taken from the onset of feeding supplemented diets at 1, 2, 3 and 4 weeks. During the LAB-feeding period, Lc. lactis ssp. lactis, Lb. sakei and Leu. mesenteroides persisted in the fish intestines, but the number of LAB slowly decreased in the intestines after changing to the unsupplemented diet. Only Lb. lactis ssp. lactis and Leu. mesenteroides were detected at levels above 1 × 102 colony-forming units/g at the end of the fourth week. In comparison to untreated control fish, the alternative complement activity in the serum was found to be significantly greater in all LAB groups at the end of the second week. Groups supplemented with Lc. lactis ssp. lactis and Leu. mesenteroides exhibited an elevated level of lysozyme activity at the end of the third week, but the group supplemented with Lb. sakei did not exhibit any significant change in lysozyme activity. Serum immunoglobulin levels were higher compared with the control group, but there was no significant difference between the LAB and control groups.

► Ex vivo transfection of pronephros cells for DNA vaccines screening. ► Successful expression of GFP and GVHSV in transfected pronephros cells. ► GVHSV up-regulated migm, tcr and tnfα 24h post-transfection. ► GVHSV up-regulated mx and tnfα, and induced an anti-VHSV state, 72h post-transfection. DNA vaccination opened a new era in controlling and preventing viral diseases since DNA vaccines have shown to be very efficacious where some conventional vaccines have failed, as it occurs in the case of the vaccines against fish novirhabdoviruses. However, there is a big lack of in vitro model assays with immune-related cells for preliminary screening of in vivo DNA vaccine candidates. In an attempt to solve this problem, rainbow trout pronephros cells in early primary culture were transfected with two plasmid DNA constructions, one encoding the green fluorescent protein (GFP) and another encoding the viral haemorrhagic septicaemia virus (VHSV) glycoprotein G (GVHSV) – the only viral antigen which has conferred in vivo protection. After assessing the presence of GFP- and GVHSV-expressing cells, at transcription and protein levels, the immune response in transfected pronephros cells was evaluated. At 24h post-transfection, GVHSV up-regulated migm and tcr transcripts expression, suggesting activation of B and T cells, as well, a high up-regulation of tnfα gene was observed. Seventy-two hours post-transfection, we detected the up-regulation of mx and tnfα genes transcripts and Mx protein which correlated with the induction of an anti-VHSV state. All together we have gathered evidence for successful transfection of pronephros cells with pAE6G, which correlates with in vivo protection results, and is less time-consuming and more rapid than in vivo assays. Therefore, this outcome opens the possibility to use pronephros cells in early primary culture for preliminary screening fish DNA vaccines as well as to further investigate the function that these cells perform in fish immune response orchestration after DNA immunisation.

The teleost fishes depend on their innate immune system for first line of defense. Mannose binding lectin (MBL) is a member of the collectin family of proteins that plays an important role in the innate immune response in many vertebrates including fish. The present study was thus carried out to identify and characterize MBL homologue from snow trout. The cDNA of snow trout MBL homologue (galactose binding; Sch-GBL) was cloned and characterized. The obtained cDNA (778-bp) contains 762 bp long ORF (open reading frame) and a 16 bp partial 3′UTR (untranslated region); transcripts are ubiquitously expressed in all the tissues under study. Sch-GBL encodes a 253-aa protein that clusters with that of common carp, goldfish and contain several common structural motifs. In addition, the K a /K s ratios of the C-type lectin domain sequences depicts that they are under negative selection. Sch-GBL transcript level increases in response to challenge to Aeromonas hydrophila infection and decreases in response to downshift in temperature. Altogether, these findings suggest that Sch-GBL play an important role in the first line of fish defense mechanisms.

Arctic charr, a highly plastic salmonid that inhabits the circumpolar region, colonized its current environment after the last glaciation. Recent colonization limits the capacity of many techniques to define and characterize constituent populations. As a novel approach, we used the major histocompatibility (MH) class IIalpha gene polymorphism as a marker that would characterize the genetic divergence of global Arctic charr populations caused by drift and by local adaptation to pathogens. We were able to detect significant isolation of all the lineages previously defined by mitochondrial DNA sequencing and also isolation of some populations within those groups. We found that most of the polymorphism of the class IIalpha gene was distributed globally, which indicates ancestral selection; however, in most cases, distinctive allele frequencies and specific haplotypes distinguished each population suggesting that recent selection has also occurred. Although all studied populations showed similar MH class IIalpha polymorphisms, we also found variation in which particular amino acid positions were polymorphic and which were constant in the different populations studied. This variation provides a greater adaptive capacity for the MH class IIalpha receptors in Arctic charr and is yet another illustration of the extraordinary plasticity of the species.