Search

Background: Growth regulation is a complex process influenced by genetic and environmental factors. We examined differences between growth hormone (GH) transgenic (T) and non-transgenic (NT) coho salmon to elucidate whether the same loci were involved in controlling body size and gene expression phenotypes, and to assess whether physiological transformations occurring from GH transgenesis were under the influence of alternative pathways. The following genomic techniques were used to explore differences between size classes within and between transgenotypes (T vs. NT): RNA-Seq/Differentially Expressed Gene (DEG) analysis, quantitative PCR (qPCR) and OpenArray analysis, Genotyping-by-Sequencing, and Genome-Wide Association Study (GWAS)., Results: DEGs identified in comparisons between the large and small tails of the size distributions of T and NT salmon (NT Large , NT Small , T Large and T Small ) spanned a broad range of biological processes, indicating wide-spread influence of the transgene on gene expression. Overexpression of growth hormone led to differences in regulatory loci between transgenotypes and size classes. Expression levels were significantly greater in T fish at 16 of 31 loci and in NT fish for 10 loci. Eleven genes exhibited different mRNA levels when the interaction of size and transgenotype was considered (IGF1, IGFBP1, GH, C3-4, FAS, FAD6, GLUT1, G6PASE1, GOGAT, MID1IP1). In the GWAS, 649 unique SNPs were significantly associated with at least one study trait, with most SNPs associated with one of the following traits: C3_4, ELA1, GLK, IGF1, IGFBP1, IGFII, or LEPTIN. Only 1 phenotype-associated SNP was found in common between T and NT fish, and there were no SNPs in common between transgenotypes when size was considered., Conclusions: Multiple regulatory loci affecting gene expression were shared between fast-growing and slow-growing fish within T or NT groups, but no such regulatory loci were found to be shared between NT and T groups. These data reveal how GH overexpression affects the regulatory responses of the genome resulting in differences in growth, physiological pathways, and gene expression in T fish compared with the wild type. Understanding the complexity of regulatory gene interactions to generate phenotypes has importance in multiple fields ranging from applications in selective breeding to quantifying influences on evolutionary processes.

The effect of dietary supplementation with a synbiotic mixture of galacto-oligosaccharides (GOS) and Bacillus spp. was examined in Caspian salmon, Salmo trutta caspius (Kessler, 1877) fingerlings. Caspian salmon fed with the synbiotic diet had significantly higher weight gain rate, protein efficiency ratio, and survival rate, as well as lower feed conversion ratio, compared to the control group (P < 0.05). The serum protein, albumin, globulin, and lactate dehydrogenase levels of the fish fed with the synbiotic diet were significantly higher than the control group (P < 0.05), while the serum alkaline phosphatase levels were significantly lower (P < 0.05). The activities of the innate immune response parameters, including lysozyme, superoxide dismutase, and catalase were significantly higher in the Caspian salmon fed with the synbiotic diet (P < 0.05). The gut microbiota of the Caspian salmon fed with the synbiotic diet contained significantly elevated total viable aerobic bacterial counts (TVABCs), lactic acid bacteria (LAB) levels, and LAB/TVABCs ratio (P < 0.05). Additionally, the gut activities of amylase, trypsin, and chymotrypsin in the gut, as well as the trypsin/chymotrypsin ratio, were significantly increased in the fish that received the synbiotic diet (P < 0.05). In conclusion, the combined GOS and Bacillus spp. supplement positively affected the growth, survival rate, immunobiochemical parameters, digestive activity, and beneficial microbial density in the gut of Caspian salmon fingerlings.

One of the categories of environmental contaminants possibly contributing to declining sockeye salmon (Oncorhynchus nerka) in the Fraser River, British Columbia, Canada is pesticides. In this 4-month study, the effects of environmentally relevant concentrations of a waterborne neonicotinoid, clothianidin (0.15, 1.5, 15 and 150 μg/L), on embryonic, alevin and early swim-up fry sockeye salmon derived from four unique genetic crosses of the Pitt River, BC stock were investigated. There were no significant effects of clothianidin on survival, hatching, growth or deformities, although genetic variation significantly affected these endpoints. Clothianidin caused a significant 4.7-fold increase in whole body 17β-estradiol levels in swim-up fry after exposure to 0.15 μg/L, but no effects were observed on testosterone levels. In addition, hepatic expression of the gene encoding glucocorticoid receptor 2 was also impacted at the highest concentration of clothianidin tested, and was found to be ∼4-fold lower compared to the sockeye reared in control water. These results indicate additional examination of clothianidin and its effects on salmonid gonad development and the reproductive and stress endocrine axes in general, is warranted., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The composition of Atlantic salmon feed has changed considerably over the last two decades from being marine-based (fishmeal and fish oil) to mainly containing plant ingredients. Consequently, concern related to traditional persistent contaminants typically associated with fish-based feed has been replaced by other potential contaminants not previously associated with salmon farming. This is the case for many pesticides, which are used worldwide to increase food production, and may be present in plant ingredients. Earlier studies have identified two organophosphorus pesticides, chlorpyrifos-methyl and pirimiphos-methyl, in plant ingredients used for aquafeed production. In the present study, we developed a reliable and sensitive analytical method, based on liquid chromatography coupled to tandem mass spectrometry, for the determination of these pesticides and their main metabolites in warm water (zebrafish) and cold water (Atlantic salmon) species, where possible differences in metabolites could be expected. The method was tested in whole zebrafish and in different salmon tissues, such as muscle, bile, kidney, fat, and liver. The final objective of this work was to assess kinetics of chlorpyrifos-methyl and pirimiphos-methyl and their main metabolites in fish tissue, in order to fill the knowledge gaps on these metabolites in fish tissues when fed over prolonged time.

Interbreeding between hatchery-reared and wild fish, through deliberate stocking or escapes from fish farms, can result in rapid phenotypic and gene expression changes in hybrids, but the underlying mechanisms are unknown. We assessed if one generation of captive breeding was sufficient to generate inter- and/or transgenerational epigenetic modifications in Atlantic salmon. We found that the sperm of wild and captive-reared males differed in methylated regions consistent with early epigenetic signatures of domestication. Some of the epigenetic marks that differed between hatchery and wild males affected genes related to transcription, neural development, olfaction, and aggression, and were maintained in the offspring beyond developmental reprogramming. Our findings suggest that rearing in captivity may trigger epigenetic modifications in the sperm of hatchery fish that could explain the rapid phenotypic and genetic changes observed among hybrid fish. Epigenetic introgression via fish sperm represents a previously unappreciated mechanism that could compromise locally adapted fish populations., (© The Author(s) 2018. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.)

A method was developed for the analysis of 22 antiparasitic residues belonging to the benzoylurea, organophosphate, pyrimidinamine, pyrethrin and pyrethroid classes in salmon by liquid chromatography coupled with tandem mass spectrometry. Samples were extracted with acetonitrile-water as the extraction solvent with use of a vibrational shaking apparatus with a ceramic homogenizer. After extraction, the acetonitrile extracts were cleaned up by incubation at low temperature (-20 °C, 1 h) to remove fat, followed by dispersive solid-phase extraction using Z-Sep+ and primary-secondary amine as sorbents. Validation was performed following the 2002/657/EC and SANTE/11813/2017 guidelines. The trueness of the method ranged from 87% to 121% and precision ranged from 4.1% to 23.7%, with the exception of cyphenothrin, dicyclanil and azamethiphos. The method developed is particularly advantageous because the use of a vibrational shaker allows unattended extraction of samples and eliminates a laborious tissue disruption step, which increases sample throughput in the laboratory. The sample preparation and chromatographic separations can be performed in 5 and 4 h, respectively, for 36 samples. Graphical abstract.

In response to public concern in Canada regarding health impacts attributable to the Fukushima Daiichi nuclear accident, oceanic seawater samples from the north Pacific and Arctic oceans, coastal seawater samples from 16 locations along the British Columbia coastline, and seafood samples (salmon, steelhead trout, and shellfish) from British Columbia coastal waters were collected and analyzed. This paper reports radiological analysis results of Pacific salmon samples (Oncorhynchus species) obtained from summer 2013 to fall 2016. While radioactive cesium from the Fukushima disaster was not detectable in most salmon samples, naturally occurring Po was measured in almost all individual samples in varying activity concentrations, from below the detection limit of 0.2 Bq kg fresh weight up to 4.7 Bq kg fresh weight. The average Po concentration among 297 salmon samples was 0.73 Bq kg fresh weight. The average ingested radiation dose per kilogram of salmon from Po is estimated to be 0.88 μSv, and the average dose from Cs is estimated to be 0.0026 μSv. The annual dose from ingested salmon would be only a fraction of the worldwide average annual effective dose from exposure to natural background radiation (2,400 μSv y) (). The measurement results showed clearly that radiation doses to people consuming fish (such as salmon) from the Canadian west coast pose no health concern.

Colour vision is mediated by the expression of different visual pigments in photoreceptors of the vertebrate retina. Each visual pigment is a complex of a protein (opsin) and a vitamin A chromophore; alterations to either component affects visual pigment absorbance and, potentially, the visual capabilities of an animal. Many species of fish undergo changes in opsin expression during retinal development. In the case of salmonid fishes the single cone photoreceptors undergo a switch in opsin expression from SWS1 (ultraviolet sensitive) to SWS2 (blue-light sensitive) starting at the yolk-sac alevin stage, around the time when they first experience light. Whether light may initiate this event or produce a plastic response in the various photoreceptors is unknown. In this study, Chinook salmon Oncorhynchus tshawytscha were exposed to light from the embryonic (5 days prior to hatching) into the yolk sac alevin (25 days post hatching) stage and the spectral phenotype of photoreceptors assessed with respect to that of unexposed controls by in situ hybridization with opsin riboprobes. Light exposure did not change the spectral phenotype of photoreceptors, their overall morphology or spatial arrangement. These results concur with those from a variety of fish species and suggest that plasticity in photoreceptor spectral phenotype via changes in opsin expression may not be a widespread occurrence among teleosts., (© 2018 The Fisheries Society of the British Isles.)

The valorization of by-products from natural organic sources is an international priority to respond to environmental and economic challenges. In this context, electrodialysis with filtration membrane (EDFM), a green and ultra-selective process, was used to separate peptides from salmon frame protein hydrolysate. For the first time, the simultaneous separation of peptides by three ultrafiltration membranes of different molecular-weight exclusion limits (50, 20, and 5 kDa) stacked in an electrodialysis system, allowed for the generation of specific cationic and anionic fractions with different molecular weight profiles and bioactivity responses. Significant decreases in peptide recovery, yield, and molecular weight (MW) range were observed in the recovery compartments depending on whether peptides had to cross one, two, or three ultrafiltration membranes. Moreover, the Cationic Recovery Compartment 1 fraction demonstrated the highest increase (42%) in glucose uptake on L6 muscle cells. While, in the anionic configuration, both Anionic Recovery Compartment 2 and Anionic Recovery Compartment 3 fractions presented a glucose uptake response in basal condition similar to the insulin control. Furthermore, Cationic Recovery Compartment 3 was found to contain inhibitory peptides. Finally, LC-MS analyses of the bioassay-guided bioactive fractions allowed us to identify 11 peptides from salmon by-products that are potentially responsible for the glucose uptake improvement.

The San Francisco Bay delta (USA) is experiencing seasonally warmer waters attributable to climate change and receives rainstorm runoff containing pyrethroid pesticides. Chinook salmon (Oncorhynchus tshawytscha) inhabit the affected waterways from hatch through smoltification, and thus juvenile fish may experience both pyrethroid and warmer water exposures. The effects of higher temperatures and pesticide exposure on presmolt Chinook are unknown. To improve understanding of the potential interaction between temperature and pesticide exposure on salmonid development, juvenile alevin and fry were reared in 11, 16.4, and 19 °C freshwater for 11 d and 2 wk, respectively, and exposed to nominal concentrations of 0, 0.15, and 1.5 µg/L bifenthrin for the final 96 h of rearing. Estradiol-17β (E2), testosterone, triiodothyronine, and thyroxine levels were measured in whole-body homogenates using hormone-specific enzyme-linked immunosorbent assays. Brain gonadotropin-releasing hormone receptor (GnRH2), dopamine receptor 2A, and growth hormone 1 (GH1) mRNA levels were measured using quantitative PCR. Results showed significantly decreased survival and condition factors observed with increasing temperature in alevin. Alevin thyroid hormones increased significantly with temperature, but fry thyroid hormones trended toward a decrease at lower temperatures with increasing bifenthrin exposure. There were significant reductions in fry testosterone and E2 at 11 °C with increasing bifenthrin treatments and significant changes in GnRH2 and GH1 gene expression in both alevin and fry, indicating potential disruption of hormonal and signaling pathways. Environ Toxicol Chem 2019;38:852-861. © 2019 SETAC., (© 2019 SETAC.)

Salmon milt extract contains high levels of nucleic acids and has antioxidant potential. Although salmon milt extract is known to improve impaired brain function in animal models with brain disease, its effects on learning and memory ability in healthy subjects is unknown. The purpose of the present study was to clarify the effect of hydrolyzed salmon milt extract (HSME) on object recognition and object location memory under normal conditions. A diet containing 2.5% HSME induced normal mice to devote more time to exploring novel and moved objects than in exploring familiar and unmoved objects, as observed during novel object recognition and spatial recognition tests, respectively. A diet containing 2.5% nucleic acid fraction purified from HSME also induced similar effects, as measured by the same behavioral tests. This suggests that the nucleic acids may be a functional component contributing to the effects of HSME on brain function. Quantitative polymerase chain reaction analysis revealed that gene expression of the markers for brain parenchymal cells, including neural stem cells, astrocytes, oligodendrocytes, and microglia, in the hippocampi of mice on an HSME diet was higher than that in mice on a control diet. Oral administration of HSME increased concentrations of cytosine, cytidine, and deoxycytidine in the hippocampus. Overall, ingestion of HSME may enhance object recognition and object location memory under normal conditions in mice, at least, in part, via the activation of brain parenchymal cells. Our results thus indicate that dietary intake of this easily ingestible food might enhance brain function in healthy individuals.

Concentration of organochlorine pesticides (OCPs) (α-, β-, γ- hexachlorocyclohexane (HCH), dichlorodiphenyltrichloroethane (DDT), dichlorodiphenyldichloroethane (DDD), dichlorodiphenyldichloroethylene (DDE)) in four species of Pacific salmon (pink, chum, chinook, and sockeye) are presented. OCPs in salmon organs increased in the following order: muscle < liver < eggs < male gonads. Concentrations of the OCP in salmon organs increased in following order: DDE < γ-HCH < α-HCH. The level of pollutants in salmon is compared with the sanitary and epidemiological norms of Russia and other countries. Cancer and noncancer hazard ratios through consumption of salmon in Russian Far East for both men and women also were summarized. Noncancer and cancer hazard ratio values were far below threshold values (<1.0)., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

A combination of previous studies and the present study indicated species-specific debromination of polybromodiphenyl ethers (PBDEs) in teleost fish. Three situations of debromination were found, namely rapid debromination represented by debromination of BDE 99 to BDE 47 observed in common carp, tilapia, crucian carp, and oscar fish; slow debromination represented by debromination of BDE 99 to BDE 49 observed in the abovementioned fish and rainbow trout, salmon, and snakehead; and no or minor debromination observed in catfish. The results of experiments on cofactors, inhibitors, and substrate competitors indicated that activities of outer ring deiodinase of 3, 3', 5'-triiodothyronine (type I deiodinase), which cannot be inhibited by 6-propyl-2-thiouracil, were responsible for the rapid debromination, and the outer ring deiodinase of thyroxine (type II deiodinase) regulated the slow debromination. The debromination of BDE 99 to BDE 49 was more common, but occurred at a much lower rate (approximately 100 times lower) than the debromination of BDE 99 to BDE 47. This was because the activity of type II deiodinase was nearly two orders of magnitude lower than that of type I deiodinase in the fish species studied. Further studies on debromination of PBDEs and properties of deiodinase in more species are needed to confirm the hypothesis., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Aeromonas salmonicida (A. salmonicida) is a pathogenic bacterium that causes furunculosis and poses a significant global risk, particularly in economic activities such as Atlantic salmon (Salmo salar) farming. In a previous study, we identified proteins that are significantly upregulated in kidneys of Atlantic salmon challenged with A. salmonicida. Phosphoproteomic analyses were conducted to further clarify the dynamic changes in protein phosphorylation patterns triggered by bacterial infection. To our knowledge, this is the first study to characterize phosphorylation events in proteins from A. salmonicida-infected Atlantic salmon. Overall, we identified over 5635 phosphorylation sites in 3112 proteins, and 1502 up-regulated and 77 down-regulated proteins quantified as a 1.5-fold or greater change relative to control levels. Based on the combined data from proteomic and motif analyses, we hypothesize that five prospective novel kinases (VRK3, GAK, HCK, PKCδ and RSK6) with common functions in inflammatory processes and cellular pathways to regulate apoptosis and the cytoskeleton could serve as potential biomarkers against bacterial propagation in fish. Data from STRING-based functional network analyses indicate that fga is the most central protein. Our collective findings provide new insights into protein phosphorylation patterns, which may serve as effective indicators of A. salmonicida infection in Atlantic salmon.

Pseudomonas fluorescens bacteria can grow well in cold-storage conditions and cause food spoilage. Quorum sensing (QS) is a biological pathway existing in a large number of microorganisms, through which bacteria regulate several of their physiological activities. A number of substances have been identified as quorum sensing inhibitors (QSIs); they can interfere with the QS system and control bacterial spoilage characteristics and production of virulence factors. In our previous study, propyl gallate at sub-minimum inhibitory concentration levels showed a potent anti-QS activity. Thus, in this study, coaxial polylactic acid-propyl gallate electrospun fibers were fabricated and their physicochemical properties were characterized. Salmon slices were coated with these electrospun fibers and the effect of this coating on the salmon slices during chilled storage was evaluated. The results showed that the electrospun fibers had a small diameter and smooth surface with no beads or other defects. The thermal stability, tensile strength, and other properties of the fibers were suitable for refrigerated storage conditions. Without inhibiting the bacterial growth in the salmon slices, the QSI-containing electrospun fibers exerted a significant inhibitory effect on the production of total volatile base nitrogen and trimethylamine. Furthermore, the deterioration of muscle tissue in the salmon slices was significantly delayed during cold storage. Quantitative analysis indicated that the electrospun fibers had a significant inhibitory effect on the bacterial spoilage ability. The results suggested that the electrospun fibers loaded with QSIs might be an effective strategy to control food spoilage and enhance the quality of aquatic food products.

Glycogen, as an intracellular deposit of polysaccharide, takes important roles in energy balance of many animals. In fish, however, the role of glycogen during development is poorly understood. In the present study, we assessed changes in glycogen concentration and gene expression patterns of glycogen-metabolizing enzymes in developing masu salmon (Oncorhynchus masou masou), a salmonid species inhabiting west side of North Pacific Ocean. As we measured glycogen levels in the bodies and yolk sacs containing the liver separately, the glycogen concentration increased in both parts as the fish developed, whereas it transiently decreased in the yolk sac after hatching, implying glycogen synthesis and breakdown in these tissues. Immunofluorescence staining using anti-glycogen monoclonal antibody revealed localization of glycogen in the liver, muscle and yolk syncytial layer of the pre-hatching embryos and hatched larvae. In order to estimate glycogen metabolism in the fish, the genes encoding homologs of glycogen synthase (gys1 and gys2) and glycogen phosphorylase (pygma, pygmb and pygl) were cloned, and their expression patterns were assessed by quantitative PCR and in situ hybridization. In the fish, gys1 and gys2 were robustly expressed in the muscle and liver, respectively. Also, expression of pyg isoforms was found in muscle, liver and yolk syncytial layer during hatching. With changes in glycogen concentration and expression patterns of relevant genes, our results suggest, for the first time, possible involvement of glycogen in energy balance of fish embryos, especially during hatching., (Copyright © 2018 Elsevier Inc. All rights reserved.)

The IsoFishR application is a data reduction and analysis tool for laser-ablation strontium isotope data, following common best practices and providing reliable and reproducible results. Strontium isotope ratios (87Sr/86Sr) are a powerful geochemical tracer commonly applied in a wide range of scientific fields and laser-ablation inductively coupled mass spectrometry is considered the method of choice to obtain spatially resolved 87Sr/86Sr isotope ratios from a variety of sample materials. However, data reduction and analyses methods are variable between different research groups and research communities limiting reproducibility between studies. IsoFishR provides a platform to standardize these methods and can be used for both spot and time-resolved line transects. Furthermore, it provides advanced data analysis tools and filters for outlier removal, noise reduction, and visualization of time resolved data. The application can be downloaded from GitHub (https://github.com/MalteWillmes/IsoFishR) and the source code is available, encouraging future development and evolution of this software., Competing Interests: The authors have declared that no competing interests exist.

Climate change increases exposure and bioaccumulation of pollutants in marine organisms, posing substantial ecophysiological and ecotoxicological risks. Here, we applied a trophodynamic ecosystem model to examine the bioaccumulation of organic mercury (MeHg) and polychlorinated biphenyls (PCBs) in a Northeastern Pacific marine food web under climate change. We found largely heterogeneous sensitivity in climate-pollution impacts between chemicals and trophic groups. Concentration of MeHg and PCBs in top predators, including resident killer whales, is projected to be amplified by 8 and 3%, respectively, by 2100 under a high carbon emission scenario (Representative Concentration Pathway 8.5) relative to a no-climate change control scenario. However, the level of amplification increases with higher carbon emission scenario for MeHg, but decreases for PCBs. Such idiosyncratic responses are shaped by the differences in bioaccumulation pathways between MeHg and PCBs, and the modifications of food web dynamics between different levels of climate change. Climate-induced pollutant amplification in mid-trophic level predators (Chinook salmon) are projected to be higher (~10%) than killer whales. Overall, the predicted trophic magnification factor is ten-fold higher in MeHg than in PCBs under high CO 2 emissions. This contribution highlights the importance of understanding the interactions with anthropogenic organic pollutants in assessing climate risks on marine ecosystems.

Objective: Various stages of mRNA processing are necessary for functionally important genes required during late-stage sperm differentiation. Protein-RNA complexes form that edit, stabilize, store, deliver, localize and regulate translation of sperm mRNAs. These regulatory processes are often directed by recognition sequence elements and the particular composition of the proteins associated with the mRNAs. Previous work has shown that the cAMP response element modulator (CREM), estrogen receptor-alpha (ERα) and forkhead box L2A (FOXL2A) proteins are present in late-stage salmon sperm. Here we investigate whether these and other regulatory proteins might control processing of mRNAs not expressed until the haploid stage of development. We also examine regulatory processes that prepare and present mRNAs that generate unique products essential for differentiating sperm (i.e. for flagellar assembly and function)., Results: We provide evidence for potential sperm-specific recognition elements in 5'-untranslated regions (utrs) that may bind CREM, ERα, FOXL2A, Y-box and other proteins. We show that changes within the 5'-utrs and open reading frames of some sperm genes lead to distinct protein termini that may provide specific interfaces necessary for localization and function within the paternal gamete.

Human activity has resulted in increasing atmospheric carbon dioxide (CO 2 ), which will result in reduced pH and higher levels of CO 2 in the ocean, a process known as ocean acidification. Understanding the effects of ocean acidification (OA) on fishes will be important to predicting and mitigating its consequences. Anadromous species such as salmonids may be especially at risk because of their rapid movements between fresh water and seawater, which could minimize their ability to acclimate. In the present study, we examine the effect of future OA on the salinity tolerance and early seawater growth of Atlantic salmon Salmo salar smolts. Exposure to 610 and 1010 μatm CO 2 did not alter salinity tolerance but did result in slightly lower plasma chloride levels in smolts exposed to seawater compared with controls (390 μatm). Gill Na + -K + -ATPase activity, plasma cortisol, glucose and haematocrit after seawater exposure were not altered by elevated CO 2 . Growth rate in the first 2 weeks of seawater exposure was greater at 1010 μatm CO 2 than under control conditions. This study of the effects of OA on S. salar during the transition from fresh water to seawater indicates that elevated CO 2 is not likely to affect osmoregulation negatively and may improve early growth in seawater., (© Published 2018. This article is a U.S. Government work and is in the public domain in the USA.)

The early life stages of Pacific salmon are at risk of environmental exposure to diluted bitumen (dilbit) as Canada's oil sands industry continues to expand. The toxicity and latent effects of dilbit exposure were assessed in sockeye salmon (Oncorhynchus nerka) exposed to water-soluble fractions of dilbit (WSFd) from fertilization to the swim-up stage, and then reared in clean water for 8 months. Mortality was significantly higher in WSFd-exposed embryos, with cumulative mortality up to 4.6-fold higher in exposed relative to unexposed embryos. The sublethal effects of WSFd exposure included transcriptional up-regulation of cyp1a, a concentration-dependent delay in the onset and progression of hatching, as well as increased prevalence of developmental deformities at total polycyclic aromatic hydrocarbon (TPAH) concentrations ≥35 μg L -1 . Growth and body composition were negatively affected by WSFd exposure, including a concentration-specific decrease in soluble protein concentration and increases in total body lipid and triglyceride concentrations. Mortality continued during the first 2 months after transferring fish to clean water, reaching 53% in fish exposed to 100 μg L -1 TPAH; but there was no latent impact on swimming performance, heart mass, or heart morphology in surviving fish after 8 months. A latent effect of WSFd exposure on brain morphology was observed, with fish exposed to 4 μg L -1 TPAH having significantly larger brains compared to other treatment groups after 8 months in clean water. This study provides comprehensive data on the acute, sub-chronic, and latent impacts of dilbit exposure in early life stage sockeye, information that is critical for a proper risk analysis of the impact of a dilbit spill on this socioeconomically important fish species., (Copyright © 2018 Elsevier B.V. All rights reserved.)

When fish perceive stressful scenarios, their hypothalamus-pituitary-interrenal axis is activated resulting in the release of corticotropin releasing hormone, adrenocorticotropic hormone (ACTH), and finally cortisol. The physiologic stress response of fish has most often been linked to the reduced performance of the immune system, with a few exceptions where the immune system is activated. In this report, we tested the hypothesis that oxidative burst activity levels in juvenile Chinook salmon (Oncorhynchus tshawytscha) are altered when the fish is presented with a stressor. Fish were subjected to a stressor for 3 h and then allowed to recover for 20 h following the stressor. Plasma and spleens were collected from euthanized fish before the stressor, at the end of a 3 h stressor, and 23 h after the start of the experiment. Plasma was held at -80 °C until cortisol radioimmunoassay analysis was performed to confirm stress. Spleens were held in Dulbecco's Modified Eagle Medium overnight and analyzed the day following collection. Oxidative burst activity was measured in splenic leukocytes after being stimulated with phorbol 12-myristate 13-acetate. We found a significant increase in activated oxidative burst from fish subjected to the stressor as compared to unstressed fish. Speculation is given to ACTH being the leukocyte priming agent in this experiment rather than the cortisol itself., (Copyright © 2018. Published by Elsevier Ltd.)

We present data from two experiments that examined how the developmental processes of smoltification and sexual maturation proceed in parallel in domesticated Atlantic salmon. Onset of maturation and smoltification was stimulated using temperature and photoperiod. Our observations on gonadosomatic index (GSI), spermatogenic activity, gill Na+, K+-ATPase enzyme (NKA) activity, and plasma 11-ketotestosterone (11-KT), Na, Cl, and Ca show that smoltification and maturation were both triggered and developed in parallel in male Atlantic salmon, but that the progressing maturation impaired hypoosmoregulation. Female maturation started after completion of smoltification. Furthermore, we present data showing that domesticated salmon can physiologically smoltify-desmoltify-resmoltify within a short period of time, and that development of a secondary sexual characteristic, such as a kype, depends on size in male postsmolts., (© 2018 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of The Physiological Society and the American Physiological Society.)

Smolting in Atlantic salmon Salmo salar is a critical life-history stage that is preparatory for downstream migration and entry to seawater that is regulated by abiotic variables including photoperiod and temperature. The present study was undertaken to determine the interaction of temperature and salinity on salinity tolerance, gill osmoregulatory proteins and cellular and endocrine stress in S. salar smolts. Fish were exposed to rapid changes in temperature (from 14 to 17, 20 and 24°C) in fresh water (FW) and seawater (SW), with and without prior acclimation and sampled after 2 and 8 days. Fish exposed simultaneously to SW and 24°C experienced 100% mortality, whereas no mortality occurred in any of the other groups. The highest temperature also resulted in poor ion regulation in SW with or without prior SW acclimation, whereas no substantial effect was observed in FW. Gill Na + -K + -ATPase (NKA) activity increased in SW fish compared to FW fish and decreased with high temperature in both FW and SW. Gill Nkaα1a abundance was high in FW and Nkaα1b and Na + -K + -2Cl- cotransporter high in SW, but all three were lower at the highest temperature. Gill Hsp70 levels were elevated in FW and SW at the highest temperature and increased with increasing temperature 2 days following direct transfer to SW. Plasma cortisol levels were elevated in SW at the highest temperature. Our results indicate that there is an important interaction of salinity and elevated temperature on osmoregulatory performance and the cellular stress response in S. salar, with an apparent threshold for osmoregulatory failure in SW above 20°C., (© 2018 The Fisheries Society of the British Isles.)

The rapid expansion of the aquaculture industry with carnivorous fish such as salmon has been accompanied by an equally rapid development in alternative feed ingredients. This has outpaced the ability of prevailing authentication method to trace the diet and origins of salmon products at the retail end. To close this gap, we developed a new profiling tool based on amino acid δ 13 C fingerprints. With this tool, we discriminated with high-accuracy among wild-caught, organically, and conventionally farmed salmon groups, as well as salmon fed alternative diets such as insects and macroalgae. Substitution of fishmeal with macroalgae was detected at 5% difference level. The δ 13 C fingerprints of essential amino acids appear particularly well suited for tracing protein sources, and the non-essentials for tracing lipid origins (terrestrial vs. aquatic). In an industry constantly developing new feed proteins and functional additives, our method is a promising tool for tracing salmon and other seafood products., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Animal and aquatic meats represent important sources of dietary protein and micro-nutrients. Although red and processed meats carry some risks for human health, sensory and nutritional advantages drive meat consumption. Therefore, it is important to understand how meat processing and cooking influence healthiness. The research aim was to investigate relationships of meat composition (proximates, amino acids and minerals) and cooking conditions (raw, 90 s microwave, 200 °C oven for 10 or 30 min) on protein digestibility, for a selection of four animal (beef, chicken, pork, kangaroo) and four aquatic meats (salmon, trout, prawn, oyster). Lean meats were minced before cooking followed by in vitro gastro-intestinal digestion and analysed for progress of hydrolysis, and size ranges of peptides using MALDI-TOF-MS. Correlation matrix analysis between compositional and functional parameters indicated that digestibility was significantly linked with protein and metal concentrations, likely reflecting moisture-dependent solubility and inter-mixing of sarcoplasmic metallo-proteins and insoluble myofibrillar proteins., (Crown Copyright © 2018. Published by Elsevier Ltd. All rights reserved.)

Curcumin (diferuloylmethane) is a natural bioactive compound with many health-promoting benefits. However, its poor water solubility and bioavailability has limited curcumin&rsquo;s biomedical application. In the present study, we encapsulated curcumin into liposomes, formed from natural sources (salmon lecithin), and characterized its encapsulation efficiency and release profile. The proposed natural carriers increased the solubility and the bioavailability of curcumin. In addition, various physico-chemical properties of the developed soft nanocarriers with and without curcumin were studied. Nanoliposome-encapsulated curcumin increased the viability and network formation in the culture of primary cortical neurons and decreased the rate of apoptosis.

Recent findings indicate that some teleost fishes may be able to greatly enhance hemoglobin-oxygen (Hb-O 2 ) unloading at the tissues under conditions that result in catecholamine release. The putative mechanism relies on the high pH sensitivity of teleost hemoglobin (Hb), intracellular red blood cell (RBC) pH regulation via β-adrenergic Na + /H + exchanger (β-NHE) activity, and plasma-accessible carbonic anhydrase at the tissues that short-circuits RBC pH regulation. Previous studies have shown that in rainbow trout, this system may double Hb-O 2 unloading to red muscle compared to a situation without short-circuiting. The present study determined that: (1) in rainbow trout this system may be functional even at low concentrations of circulating catecholamines, as shown by conducting a dose-response analysis; (2) Atlantic and coho salmon also possess β-NHE activity, as shown by changes in hematocrit in adrenergically stimulated cells; and (3) with β-NHE short-circuiting, Atlantic and coho salmon may be able to increase Hb-O 2 unloading by up to 74 and 159%, respectively, as determined by modeling based on O 2 equilibrium curves. Together, these results indicate that a system to enhance Hb-O 2 unloading may be common among salmonids and may be operational even under routine conditions. In view of the life histories of Atlantic and coho salmon, a system to enhance Hb-O 2 unloading during exercise may help determine a successful spawning migration and thus reproductive success.

Several metabolic parameters were assessed in juvenile Chinook salmon (Oncorhynchus tshawytscha) and staghorn sculpin (Leptocottus armatus) residing in two estuaries receiving wastewater treatment effluent and one reference estuary. We also conducted a laboratory study with fish dosed for 32 days with 16 of the most common contaminants of emerging concern (CECs) detected in feral fish. Several blood chemistry parameters and other indicators of health were measured in fish from the field and laboratory study that were used to assess potential metabolic disruption. The blood chemistry values observed in feral juvenile Chinook salmon were relatively consistent among fish collected from effluent-impacted sites and substantially different compared to reference site fish. These responses were more pronounced in Chinook salmon, which is supported by the disparity in accumulated CECs. The blood chemistry results for juvenile Chinook salmon collected at effluent-impacted sites exhibited a pattern generally consistent with starvation because of similarities to observations from studies of food-deprived fish; however, this response is not consistent with physical starvation but may be contaminant induced. The altered blood chemistry parameters are useful as an early indicator of metabolic stress, even though organismal characteristics (lipid content and condition factor) were not different among sites indicating an early response. Evidence of metabolic disruption was also observed in juvenile Chinook salmon that were exposed in the laboratory to a limited mixture of CECs; however, the plasma parameters were qualitatively different possibly due to exposure route, season, or the suite of CECs. Growth was impaired in the high-dose fish during the dosing phase and the low- and medium-dose fish assayed after 2 weeks of depuration. Overall, these results are consistent with metabolic disruption for fish exposed to CECs, which may result in early mortality or an impaired ability to compete for limited resources., (Published by Elsevier Ltd.)

Benzo[a]pyrene (BaP), a polycyclic aromatic hydrocarbon (PAH), is a known human carcinogen. In non-smoking adults greater than 95% of BaP exposure is through diet. The carcinogenicity of BaP is utilized by the U.S. EPA to assess relative potency of complex PAH mixtures. PAH relative potency factors (RPFs, BaP = 1) are determined from high dose animal data. We employed accelerator mass spectrometry (AMS) to determine pharmacokinetics of [ 14 C]-BaP in humans following dosing with 46 ng (an order of magnitude lower than human dietary daily exposure and million-fold lower than animal cancer models). To assess the impact of co-administration of food with a complex PAH mixture, humans were dosed with 46 ng of [ 14 C]-BaP with or without smoked salmon. Subjects were asked to avoid high BaP-containing diets and a 3-day dietary questionnaire given to assess dietary exposure prior to dosing and three days post-dosing with [ 14 C]-BaP. Co-administration of smoked salmon, containing a complex mixture of PAHs with an RPF of 460 ng BaP eq , reduced and delayed absorption. Administration of canned commercial salmon, containing very low amounts of PAHs, showed the impacts on pharmacokinetics were not due to high amounts of PAHs but rather a food matrix effect., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Fourier Transform infrared imaging and multivariate analysis were used to identify, at the microscopic level, the presence of florfenicol (FF), a heavily-used antibiotic in the salmon industry, supplied to fishes in feed pellets for the treatment of salmonid rickettsial septicemia (SRS). The FF distribution was evaluated using Principal Component Analysis (PCA) and Augmented Multivariate Curve Resolution with Alternating Least Squares (augmented MCR-ALS) on the spectra obtained from images with pixel sizes of 6.25 μm × 6.25 μm and 1.56 μm × 1.56 μm, in different zones of feed pellets. Since the concentration of the drug was 3.44 mg FF/g pellet, this is the first report showing the powerful ability of the used of spectroscopic techniques and multivariate analysis, especially the augmented MCR-ALS, to describe the FF distribution in both the surface and inner parts of feed pellets at low concentration, in a complex matrix and at the microscopic level. The results allow monitoring the incorporation of the drug into the feed pellets., (Copyright © 2018 Elsevier B.V. All rights reserved.)

'Are we what we eat?' Yes and no. Although dietary fat affects body fat, there are many modifying mechanisms. In Atlantic salmon, there is a high level of retention of the n-3 fatty acid (FA) docosahexaenoic acid (DHA, 22:6n-3) relative to the dietary content, whereas saturated FAs never seem to increase above a specified level, which is probably an adaptation to low and fluctuating body temperature. Net production of eicosapentaenoic acid (EPA, 20:5n-3) and especially DHA occurs in salmon when dietary levels are low; however, this synthesis is not sufficient to maintain EPA and DHA at similar tissue levels to those of a traditional fish oil-fed farmed salmon. The commercial diets of farmed salmon have changed over the past 15 years towards a more plant-based diet owing to the limited availability of the marine ingredients fish meal and fish oil, resulting in decreased EPA and DHA and increased n-6 FAs. Salmon is part of the human diet, leading to the question 'Are we what the salmon eats?' Dietary intervention studies using salmon have shown positive effects on FA profiles and health biomarkers in humans; however, most of these studies used salmon that were fed high levels of marine ingredients. Only a few human intervention studies and mouse trials have explored the effects of the changing feed composition of farmed salmon. In conclusion, when evaluating feed ingredients for farmed fish, effects throughout the food chain on fish health, fillet composition and human health need to be considered., Competing Interests: Competing interestsThe author declares no competing or financial interests., (© 2018. Published by The Company of Biologists Ltd.)

Given the compound differences between tris(2-butoxyethyl)- and tris(2-cloroethyl) phosphate (TBOEP and TCEP, respectively), we hypothesized that exposure of juvenile salmon to TBOEP and TCEP will produce compound-specific differences in uptake and bioaccumulation patterns, resulting in potential formation of OH-metabolites. Juvenile salmon were exposed to waterborne TCEP or TBOEP (0.04, 0.2 and 1 mg/L) for 7 days. The muscle accumulation was measured and bioconcentration factor (BCF) was calculated, showing that TCEP was less accumulative and resistant to metabolism in salmon than TBOEP. Metabolite formations were only detected in TBOEP-exposed fish, showing seven phase I biotransformation metabolites with hydroxylation, ether cleavage or combination of both reactions as important metabolic pathways. In vitro incubation of trout S9 liver fraction with TBOEP was performed showing that the generated metabolite patterns were similar to those found in muscle tissue exposed in vivo. However, another OH-TBOEP isomer and an unidentified metabolite not present in in vivo exposure were observed with the trout S9 incubation. Overall, some of the observed metabolic products were similar to those in a previous in vitro report using human liver microsomes and some metabolites were identified for the first time in the present study. Toxicological analysis indicated that TBOEP produced less effect, although it was taken up faster and accumulated more in fish muscle than TCEP. TCEP produced more severe toxicological responses in multiple fish organs. However, liver biotransformation responses did not parallel the metabolite formation observed in TBOEP-exposed fish., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Although the keto-carotenoid astaxanthin (Ast) is not typically present in human plasma due to its relative scarcity in the typical diet, global consumption of salmon, the primary source of Ast in food, and Ast supplements continues to increase. The first objective of the present study was to investigate the bioaccessibility of Ast from uncooked and cooked fillets of wild and aquacultured salmon, Ast-supplements and krill oil, during simulated gastric and small intestinal digestion. Uptake of E-Ast from micelles generated during digestion of wild salmon by monolayers of Caco-2 was also monitored. Both wild and aquacultured salmon flesh contained E-Ast and Z-isomers of unesterified Ast, whereas Ast esters were the predominant form of the carotenoid in commercial supplements and krill oil. Flesh from wild salmon contained approximately 10 times more Ast than aquacultured salmon. Common styles of cooking flesh from wild and aquacultured salmon decreased Ast content by 48-57% and 35-47%, respectively. Ast in salmon flesh, supplements and krill oil was relatively stable (>80% recovery) during in vitro digestion. The efficiency of transfer of Ast into mixed micelles during digestion of uncooked wild salmon was 43%, but only 12% for uncooked acquacultured salmon. Cooking wild salmon significantly decreased Ast bioaccessibility. The relative bioaccessibility of Ast (41-67%) after digestion of oil vehicle in commercial supplements was inversely proportional to carotenoid content (3-10mg/capsule), whereas bioaccessibility of endogenous Ast in phospholipid-rich krill oil supplement was 68%. >95% of Ast in mixed micelles generated during digestion of supplements and krill oil was unesterified. Caco-2 intestinal cells accumulated 11-14% of E-Ast delivered in mixed micelles generated from digested wild salmon. Apical uptake and basolateral secretion of E-Ast by Caco-2 cells grown on inserts were greater after digestion of Ast-enriched krill oil compared to uncooked wild salmon. These data suggest that the bioacessibility of Ast in wild salmon and soft-gel capsules is greater than that in aquacultured salmon, and that uptake and basolateral secretion of the carotenoid by enterocyte-like cells is enhanced by the digestion products of phospholipid-rich krill oil., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

We previously reported the bioaccumulation of contaminants of emerging concern (CECs), including pharmaceuticals and personal care products (PPCPs) and perfluorinated compounds, in field-collected juvenile Chinook salmon from urban estuaries of Puget Sound, WA (Meador et al., 2016). Although the toxicological impacts of CECs on salmon are poorly understood, several of the detected contaminants disrupt mitochondrial function in other species. Here, we sought to determine whether environmental exposures to CECs are associated with hepatic mitochondrial dysfunction in juvenile Chinook. Fish were exposed in the laboratory to a dietary mixture of 16 analytes representative of the predominant CECs detected in our field study. Liver mitochondrial content was reduced in fish exposed to CECs, which occurred concomitantly with a 24-32% reduction in expression of peroxisome proliferator-activated receptor (PPAR) Y coactivator-1a (pgc-1α), a positive transcriptional regulator of mitochondrial biogenesis. The laboratory exposures also caused a 40-70% elevation of state 4 respiration per unit mitochondria, which drove a 29-38% reduction of efficiency of oxidative phosphorylation relative to controls. The mixture-induced elevation of respiration was associated with increased oxidative injury as evidenced by increased mitochondrial protein carbonyls, elevated expression of glutathione (GSH) peroxidase 4 (gpx4), a mitochondrial-associated GSH peroxidase that protects against lipid peroxidation, and reduction of mitochondrial GSH. Juvenile Chinook sampled in a WWTP effluent-impacted estuary with demonstrated releases of CECs showed similar trends toward reduced liver mitochondrial content and elevated respiratory activity per mitochondria (including state 3 and uncoupled respiration). However, respiratory control ratios were greater in fish from the contaminated site relative to fish from a minimally-polluted reference site, which may have been due to differences in the timing of exposure to CECs under laboratory and field conditions. Our results indicate that exposure to CECs can affect both mitochondrial quality and content, and support the analysis of mitochondrial function as an indicator of the sublethal effects of CECs in wild fish., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The pancreatic hormone amylin and its agonist salmon calcitonin (sCT) act via the area postrema (AP) and the lateral parabrachial nucleus (PBN) to reduce food intake. Investigations of amylin and sCT signaling in the ventral tegmental area (VTA) and nucleus accumbens (NAc) suggest that the eating inhibitory effect of amylin is, in part, mediated through the mesolimbic 'reward' pathway. Indeed, administration of the sCT directly to the VTA decreased phasic dopamine release (DA) in the NAc. However, it is not known if peripheral amylin modulates the mesolimbic system directly or whether this occurs via the AP and PBN. To determine whether and how peripheral amylin or sCT affect mesolimbic reward circuitry we utilized fast scan cyclic voltammetry under anesthesia to measure phasic DA release in the NAc evoked by electrical stimulation of the VTA in intact, AP lesioned and bilaterally PBN lesioned rats. Amylin (50μg/kg i.p.) did not change phasic DA responses compared to saline control rats. However, sCT (50μg/kg i.p.) decreased evoked DA release to VTA-stimulation over 1h compared to saline treated control rats. Further investigations determined that AP and bilateral PBN lesions abolished the ability of sCT to suppress evoked phasic DA responses to VTA-stimulation. These findings implicate the AP and the PBN as important sites for peripheral sCT to decrease evoked DA release in the NAc and suggest that these nuclei may influence hedonic and motivational processes to modulate food intake., (Copyright © 2017 Elsevier Inc. All rights reserved.)

The saxitoxin-group (STX-group) corresponds to toxic metabolites produced by cyanobacteria and dinoflagellates of the genera Alexandrium, Gymnodinium , and Pyrodinium . Over the last decade, it has been possible to extrapolate the areas contaminated with the STX-group worldwide, including Chile, a phenomenon that has affected ≈35% of the Southern Pacific coast territory, generating a high economic impact. The objective of this research was to study the toxicity of the STX-group in all aquatic organisms (bivalves, algae, echinoderms, crustaceans, tunicates, cephalopods, gastropods, and fish) present in areas with a variable presence of harmful algal blooms (HABs). Then, the toxic profiles of each species and dose of STX equivalents ingested by a 60 kg person from 400 g of shellfish were determined to establish the health risk assessment. The toxins with the highest prevalence detected were gonyautoxin-4/1 (GTX4/GTX1), gonyautoxin-3/2 (GTX3/GTX2), neosaxitoxin (neoSTX), decarbamoylsaxitoxin (dcSTX), and saxitoxin (STX), with average concentrations of 400, 2800, 280, 200, and 2000 µg kg -1 respectively, a species-specific variability, dependent on the evaluated tissue, which demonstrates the biotransformation of the analogues in the trophic transfer with a predominance of α-epimers in all toxic profiles. The identification in multiple vectors, as well as in unregulated species, suggests that a risk assessment and risk management update are required; also, chemical and specific analyses for the detection of all analogues associated with the STX-group need to be established., Competing Interests: The authors declare no conflict of interest.

The effects of 2 currently used commercial pesticide formulations on Pacific sockeye salmon (Oncorhynchus nerka), from fertilization to emergence, were evaluated in a gravel-bed flume incubator that simulated a natural streambed. Embryos were exposed to atrazine at 25 µg/L (low atrazine) or atrazine at 250 µg/L (high atrazine) active ingredient (a.i.), and chlorothalonil at 0.5 µg/L (low chlorothalonil) or chlorothalonil at 5 µg/L a.i. (high chlorothalonil) and examined for effects on developmental success and timing, as well as physical and biochemical growth parameters. Survival to hatch was reduced in the high chlorothalonil group (55% compared with 83% in controls), accompanied by a 24% increase in finfold deformity incidence. Reduced alevin condition factor (2.9-5.4%) at emergence and elevated triglyceride levels were seen in chlorothalonil-exposed fish. Atrazine exposure caused premature hatch (average high atrazine time to 50% hatch [H50] = 100 d postfertilization [dpf]), and chlorothalonil exposure caused delayed hatch (high chlorothalonil H50 = 108 dpf; controls H50 = 102 dpf). All treatments caused premature emergence (average time to 50% emergence [E50]: control E50 = 181 dpf, low chlorothalonil E50 = 175 dpf, high chlorothalonil E50 = 174 dpf, high atrazine E50 = 175 dpf, low atrazine E50 = 174 dpf), highlighting the importance of using a gravel-bed incubator to examine this subtle, but critical endpoint. These alterations indicate that atrazine and chlorothalonil could affect survival of early life stages of sockeye salmon in the wild. Environ Toxicol Chem 2017;36:1354-1364. © 2017 SETAC., (© 2017 SETAC.)

There is a great interest in finding alternatives and green solvents in extraction processes to replace petroleum based solvents. In order to investigate these possibilities, computational methods, as Hansen solubility parameters (HSP) and conductor-like screening model for real solvent (COSMO-RS), were used in this work to predict the solvation power of a series of solvents in salmon fish lipids. Additionally, experimental studies were used to evaluate the performance in lipids extraction using 2-methyltetrahydrofurane, cyclopentyl methyl ether, dimethyl carbonate, isopropanol, ethanol, ethyl acetate, p-cymene and d-limonene compared with hexane. Lipid classes of extracts were obtained by using high performance thin-layer chromatography (HPTLC), whereas gas chromatography with a flame ionization detector (GC/FID) technique was employed to obtain fatty acid profiles. Some differences between theoretical and experimental results were observed, especially regarding the behavior of p-cymene and d-limonene, which separate from the predicted capability. Results obtained from HPTLC indicated that p-cymene and d-limonene extract triglycerides (TAGs) and diglycerides (DAGs) at levels of 73 and 19%, respectively, whereas the other studied extracts contain between 75 and 76% of TAGs and between 16 and 17% of DAGs. Fatty acid profiles, obtained by using GC-FID, indicated that saturated fatty acids (SFAs) between 19.5 and 19.9% of extracted oil, monounsaturated fatty acids (MUFAs) in the range between 43.5 and 44.9%, and PUFAs between 31.2 and 34.6% were extracted. p-Cymene and limonene extracts contained lower percentages than the other studied solvents of some PUFAs due probably to the fact that these unsaturated fatty acids are more susceptible to oxidative degradation than MUFAs. Ethyl acetate has been found to be the best alternative solvent to hexane for the extraction of salmon oil lipids. Graphical Abstract ᅟ.

This research investigated how ploidy level (diploid versus triploid) affects the heat shock protein (HSP) response in erythrocytes under different thermal stress regimes, both in vivo and in vitro, in Atlantic salmon (Salmo salar) and brook charr (Salvelinus fontinalis) in order to address the question of why triploids typically have reduced thermal tolerance. A preliminary study confirmed that identical volumes of diploid and triploid erythrocytes (which equates to a smaller number of larger cells for triploids compared to diploids) did not differ in total protein synthesis rates. After chronic (100d) acclimation of fish to 5, 15 and 25°C, triploid erythrocytes had lower HSP70, HSP90, heat shock factor 1 (HSF1) and ubiquitin (free and total) levels than diploids in both species. Furthermore, Atlantic salmon erythrocytes showed significantly higher protein breakdown (based on conjugated ubiquitin levels) in triploids than diploids after acute heat stress in vitro, but no significant difference was detected between ploidies after acute cold stress. These results indicate that: 1) triploid erythrocytes synthesize more total protein per cell than diploids as a result of increased cell size; 2) triploids have sufficient total HSP levels for survival under low stress conditions; and 3) the lower basal titres of HSPs in triploids may be a handicap when combating acute stress. Taken together, this suggests that triploids are limited in their ability to withstand thermal stress because of a reduced ability to maintain proteostasis under stressful conditions., (Copyright © 2017 Elsevier Inc. All rights reserved.)

A new technology employing Raman spectroscopy is attracting attention as a powerful biochemical technique for the detection of beneficial and functional food nutrients, such as carotenoids and unsaturated fatty acids. This technique allows for the dynamic characterization of food nutrient substances for the rapid determination of food quality. In this study, we attempt to detect and measure astaxanthin from salmon fillets using this technology. The Raman spectra showed specific bands corresponding to the astaxanthin present in salmon and the value of astaxanthin (Raman band, 1518 cm -1 ) relative to those of protein/lipid (Raman band, 1446 cm -1 ) in the spectra increased in a dose-dependent manner. A standard curve was constructed by the standard addition method using astaxanthin as the reference standard for its quantification by Raman spectroscopy. The calculation formula was established using the Raman bands typically observed for astaxanthin (i.e., 1518 cm -1 ). In addition, we examined salmon fillets of different species (Atlantic salmon, coho salmon, and sockeye salmon) and five fillets obtained from the locations (from the head to tail) of an entire Atlantic salmon. Moreover, the sockeye salmon fillet exhibited the highest astaxanthin concentration (14.2 mg/kg), while coho salmon exhibited an intermediate concentration of 7.0 mg/kg. The Raman-based astaxanthin concentration in the five locations of Atlantic salmon was more strongly detected from the fillet closer to the tail. From the results, a rapid, convenient Raman spectroscopic method was developed for the detection of astaxanthin in salmon fillets.

Chinook salmon (Oncorhynchus tshawytscha) are external fertilizers that display sneak-guard alternative reproductive tactics. The larger hooknose males dominate mating positions, while the smaller jack males utilize sneak tactics to achieve fertilization. Although poorly understood, previous studies have suggested that differences in spermatozoa quality may play a critical role in sperm competition. Considering that the seminal plasma strongly regulates spermatozoa quality and other processes critical for fertilization success, we employed label free quantitative mass spectrometry utilizing ion mobility separation coupled to cross-species bioinformatics to examine the seminal plasma proteome of Chinook salmon. A total of 345 proteins were identified in all biological replicates analyzed, including many established seminal plasma proteins that may serve as future biomarkers for Chinook salmon fertility and sperm competition. Moreover, we elucidated statistically significant protein abundance differences between hooknose and jack male tactics. Proteins involved in membrane remodeling, proteolysis, hormonal transport, redox regulation, immunomodulation, and ATP metabolism were among the proteins reproducibly identified at different levels and represent putative factors influencing sperm competition between jack and hooknose males. This study represents the largest seminal plasma proteome from teleost fish and the first reported for Chinook salmon., Significance: Chinook salmon (Oncorhynchus tshawytscha) males represent an example of male alternative reproductive tactics where diverse reproductive strategies are thought to increase sexual selection. While seminal plasma has been shown to play an important regulatory role in sperm competition in many species, little is known about the protein composition of the seminal plasma of salmon. Therefore, seminal plasma isolated from the two alternative reproductive tactics of Chinook salmon (small sneaky jacks and large dominant hooknoses) were analyzed by label free quantitative mass spectrometry employing data independent acquisition and ion mobility separation. This yielded the largest proteome data set of the seminal plasma from salmon and the first to examine protein abundance differences between male alternative reproductive tactics. The quantitative proteomic data provides insight into possible unique mechanistic aspects of Chinook salmon alternative reproductive tactics utilized for sperm competition and fertilization success., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Triploid fish has become an important item of commercial aquaculture, but data on its fatty acid (FA) composition are still controversial, especially regarding essential polyunsaturated fatty acids, eicosapentaenoic acid (20:5n-3, EPA) and docosahexaenoic acid (22:6n-3, DHA). We studied FA composition and content of diploid and triploid pink salmon Oncorhynchus gorbuscha, reared in aquaculture in a bay of the White Sea (Russia). FA composition, measured as percentages of total FA of triploids and immature diploid females significantly differed from that of mature diploid fish. Specifically, mature diploids had higher percentage of EPA and DHA in their muscle tissue (filets) compared to that of triploids and immature diploid females. Nevertheless, the contents of EPA and DHA per mass of the filets in diploid and triploid specimens were similar. Thus, no special efforts are needed to improve EPA and DHA contents in filets of triploids., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Phospholipids and their partial hydrolysates, namely lysophospholipids (LPLs), have been widely used in food, pharmaceutical, and cosmetic products as highly efficient emulsifiers. This study was conducted to produce docosahexaenoic acid (DHA)-esterified LPLs by enzymatic modification of phospholipids obtained from the head of autumn chum salmon (Oncorhynchus keta). The emulsifying properties of the obtained LPLs were also evaluated. Two different types of substrates of salmon head phospholipids were prepared via silica gel and cold acetone precipitation. Enzymatic partial hydrolysis was carried out using immobilized phospholipase A 1 (PLA 1 ) and Lipozyme RM IM. Results showed that the increase in DHA in the LPLs was much higher in the silica-separated phospholipids than in the acetone-precipitated phospholipids. When silica-separated phospholipids were used as the substrate, the DHA content of the LPLs increased from 23.1% to 40.6% and 42.6% after 8 h of partial hydrolysis with Lipozyme RM IM and immobilized PLA 1 , respectively. The yield of the LPLs was comparatively higher in the Lipozyme RM IM than in the immobilized PLA 1 hydrolysis reaction. The critical micelle concentration values of the LPLs and purified lysophosphatidylcholine (LPC) were 100 mg/L and 5 mg/L, respectively. The surface tension values of the LPLs and LPC were reduced to 30.0 mN/m and 30.5 mN/m, respectively. The hydrophilic-lipophilic balance of the LPLs and LPC were 6.0 and 9.4, respectively. Based on the emulsifying properties observed, we conclude that LPLs derived from the phospholipids of salmon head lipids could be used as a health-beneficial emulsifier in the food industry.

Diluted bitumen (dilbit; the product of oil sands extraction) is transported through freshwater ecosystems critical to Pacific salmon. This is concerning, because crude oil disrupts cardiac development, morphology, and function in embryonic fish, and cardiac impairment in salmon can have major consequences on migratory success and fitness. The sensitivity of early life-stage salmon to dilbit and its specific cardiotoxic effects are unknown. Sockeye salmon parr were exposed to environmentally relevant concentrations of the water-soluble fraction (WSF) of dilbit for 1 wk and 4 wk, followed by an examination of molecular, morphological, and organismal endpoints related to cardiotoxicity. We show that parr are sensitive to WSF of dilbit, with total polycyclic aromatic hydrocarbon (PAH) concentrations of 3.5 µg/L sufficient to induce a liver biomarker of PAH exposure, and total PAH of 16.4 µg/L and 66.7 µg/L inducing PAH biomarkers in the heart. Furthermore, WSF of dilbit induces concentration-dependent cardiac remodeling coincident with performance effects: fish exposed to 66.7 µg/L total PAH have relatively fewer myocytes and more collagen in the compact myocardium and impaired swimming performance at 4 wk, whereas the opposite changes occur in fish exposed to 3.5 µg/L total PAH. The results demonstrate cardiac sensitivity to dilbit exposure that could directly impact sockeye migratory success. Environ Toxicol Chem 2017;36:354-360. © 2016 SETAC., (© 2016 SETAC.)

Post-reproductive lifespan varies greatly among species; human post-reproductive lifespan comprises ~30-50% of their total longevity, while semelparous salmon and dasyurid marsupials post-reproductive lifespan comprises <4% of their total longevity. To examine if the magnitude of hypothalamic-pituitary-gonadal (HPG) axis dyscrasia at the time of reproductive senescence determines post-reproductive lifespan, we examined the difference between pre- and post-reproductive (1) circulating sex hormones and (2) the ratio of sex steroids to gonadotropins (e.g., 17β-estradiol/follicle-stimulating hormone (FSH)), an index of the dysregulation of the HPG axis and the level of dyotic (death) signaling post-reproduction. Animals with a shorter post-reproductive lifespan (<4% total longevity) had a more marked decline in circulating sex steroids and corresponding elevation in gonadotropins compared to animals with a longer post-reproductive lifespan (30-60% total longevity). In semelparous female salmon of short post-reproductive lifespan (1%), these divergent changes in circulating hormone concentration post-reproduction equated to a 711-fold decrease in the ratio of 17β-estradiol/FSH between the reproductive and post-reproductive periods. In contrast, the decrease in the ratio of 17β-estradiol/FSH in iteroparous female mammals with long post-reproductive lifespan was significantly less (1.7-34-fold) post-reproduction. Likewise, in male semelparous salmon, the decrease in the ratio of testosterone/FSH (82-fold) was considerably larger than for iteroparous species (1.3-11-fold). These results suggest that (1) organisms with greater reproductive endocrine dyscrasia more rapidly undergo senescence and die, and (2) the contribution post-reproduction by non-gonadal (and perhaps gonadal) tissues to circulating sex hormones dictates post-reproductive tissue health and longevity. In this way, reproduction and longevity are coupled, with the degree of non-gonadal tissue hormone production dictating the rate of somatic tissue demise post-reproduction and the differences in post-reproductive lifespans between species.

Bacterial extracellular proteases from six strains of marine bacteria and seven strains of terrestrial bacteria were prepared through fermentation. Proteases were analyzed through substrate immersing zymography and used to hydrolyze the collagen and muscle proteins from a salmon skin byproduct, respectively. Collagen could be degraded much more easily than muscle protein, but it commonly showed weaker antioxidant capability. The hydrolysate of muscle proteins was prepared with crude enzymes from Pseudoalteromonas sp. SQN1 displayed the strongest activity of antioxidant in DPPH and hydroxyl radical scavenging assays (74.06% ± 1.14% and 69.71% ± 1.97%), but did not perform well in Fe 2+ chelating assay. The antioxidant fractions were purified through ultrafiltration, cation exchange chromatography, and size exclusion chromatography gradually, and the final purified fraction U2-S2-I displayed strong activity of antioxidant in DPPH, hydroxyl radical scavenging assays (IC 50 = 0.263 ± 0.018 mg/mL and 0.512 ± 0.055 mg/mL), and oxygen radical absorption capability assay (1.960 ± 0.381 mmol·TE/g). The final purified fraction U2-S2-I possessed the capability to protect plasmid DNA against the damage of hydroxyl radical and its effect was similar to that of the original hydrolysis product. It indicated that U2-S2-I might be the major active fraction of the hydrolysate. This study proved that bacterial extracellular proteases could be utilized in hydrolysis of a salmon byproduct. Compared with collagen, muscle proteins was an ideal material used as an enzymatic substrate to prepare antioxidant peptides., Competing Interests: The authors declare no conflict of interest.

The present study aimed at investigating cytotoxicity and oxidative stress induced by silica-coated iron oxide nanoparticles functionalized with dithiocarbamate (Fe 3 O 4 NPs) in Chinook salmon cells (CHSE-214) derived from Oncorhynchus tshawytscha embryos. A significant reduction in cell viability was evident in response to Fe 3 O 4 NPs as revealed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay after 24 h of exposure. Out of the tested concentrations (10, 20, and 30 μg/ml), the highest concentration has shown significant decrease in the viability of cells after 24 h of exposure. Alterations in the morphology of CHSE-214 cells was also evident at 10 μg/ml concentration of Fe 3 O 4 NPs after 24 h. Fe 3 O 4 NPs elicited a significant dose-dependent reduction in total glutathione content (TGSH), catalase (CAT), glutathione reductase (GR) with a concomitant increase in lipid peroxidation (LPO), and protein carbonyl (PC) at highest concentration (30 μg/ml) after 24 h of exposure. In conclusion, our data demonstrated that Fe 3 O 4 NPs have potential to induce cytotoxicity in CHSE-214 cells, which is likely to be mediated through reactive oxygen species generation and oxidative stress.

Dietary nucleotides play a role in maintaining the immune responses of both animals and humans. Oral administration of nucleic acids from salmon milt have physiological functions in the cellular metabolism, proliferation, differentiation, and apoptosis of human small intestinal epithelial cells. In this study, we examined the effects of DNA-rich nucleic acids prepared from salmon milt (DNSM) on the development of liver fibrosis in an in vivo ethanol-carbon tetrachloride cirrhosis model. Plasma aspartate transaminase and alanine transaminase were significantly less active in the DNSM-treated group than in the ethanol plus carbon tetrachloride (CCl₄)-treated group. Collagen accumulation in the liver and hepatic necrosis were observed histologically in ethanol plus CCl₄-treated rats; however, DNSM-treatment fully protected rats against ethanol plus CCl₄-induced liver fibrosis and necrosis. Furthermore, we examined whether DNSM had a preventive effect against alcohol-induced liver injury by regulating the cytochrome p450 2E1 (CYP2E1)-mediated oxidative stress pathway in an in vivo model. In this model, CYP2E1 activity in ethanol plus CCl₄-treated rats increased significantly, but DNSM-treatment suppressed the enzyme's activity and reduced intracellular thiobarbituric acid reactive substances (TBARS) levels. Furthermore, the hepatocytes treated with 100 mM ethanol induced an increase in cell death and were not restored to the control levels when treated with DNSM, suggesting that digestive products of DNSM are effective for the prevention of alcohol-induced liver injury. Deoxyadenosine suppressed the ethanol-induced increase in cell death and increased the activity of alcohol dehydrogenase. These results suggest that DNSM treatment represents a novel tool for the prevention of alcohol-induced liver injury., Competing Interests: The authors declare no conflict of interest.