Species Difference? Bovine, Trout, and Human Plasma Protein Binding of Per- and Polyfluoroalkyl Substances.

Per- and polyfluoroalkyl substances (PFAS) strongly bind to proteins and lipids in blood, which govern their accumulation and distribution in organisms. Understanding the plasma binding mechanism and species differences will facilitate the quantitative in vitro-to-in vivo extrapolation and improve risk assessment of PFAS. We studied the binding mechanism of 16 PFAS to bovine serum albumin (BSA), trout, and human plasma using solid-phase microextraction. Binding of anionic PFAS to BSA and human plasma was found to be highly concentration-dependent, while trout plasma binding was linear for the majority of the tested PFAS. At a molar ratio of PFAS to protein ν < 0.1 molPFAS/molprotein, the specific protein binding of anionic PFAS dominated their human plasma binding. This would be the scenario for physiological conditions (ν < 0.01), whereas in in vitro assays, PFAS are often dosed in excess (ν > 1) and nonspecific binding becomes dominant. BSA was shown to serve as a good surrogate for human plasma. As trout plasma contains more lipids, the nonspecific binding to lipids affected the affinities of PFAS for trout plasma. Mass balance models that are parameterized with the protein-water and lipid-water partitioning constants (chemical characteristics), as well as the protein and lipid contents of the plasma (species characteristics), were successfully used to predict the binding to human and trout plasma.

The late-evolving salmon and trout join the GnRH1 club.

Although it is known that the whitefish, an ancient salmonid, expresses three distinct gonadotropin-releasing hormone (GnRH) forms in the brain, it has been thought that the later-evolving salmonids (salmon and trout) had only two types of GnRH: GnRH2 and GnRH3. We now provide evidence for the expression of GnRH1 in the gonads of Atlantic salmon by rapid amplification of cDNA ends, real-time quantitative PCR and immunohistochemistry. We examined six different salmonid genomes and found that each assembly has one gene that likely encodes a viable GnRH1 prepropeptide. In contrast to both functional GnRH2 and GnRH3 paralogs, the GnRH1 homeolog can no longer express the hormone. Furthermore, the viable salmonid GnRH1 mRNA is composed of only three exons, rather than the four exons that build the GnRH2 and GnRH3 mRNAs. Transcribed gnrh1 is broadly expressed (in 17/18 tissues examined), with relative abundance highest in the ovaries. Expression of the gnrh2 and gnrh3 mRNAs is more restricted, primarily to the brain, and not in the gonads. The GnRH1 proximal promoter presents composite binding elements that predict interactions with complexes that contain diverse cell fate and differentiation transcription factors. We provide immunological evidence for GnRH1 peptide in the nucleus of 1-year-old type A spermatogonia and cortical alveoli oocytes. GnRH1 peptide was not detected during other germ cell or reproductive stages. GnRH1 activity in the salmonid gonad may occur only during early stages of development and play a key role in a regulatory network that controls mitotic and/or meiotic processes within the germ cell.

Interspecies Differences in 6PPD-Quinone Toxicity Across Seven Fish Species: Metabolite Identification and Semiquantification.

N-(1,3-Dimethylbutyl)-N'-phenyl-p-phenylenediamine-quinone (6PPD-Q) is a recently identified contaminant that originates from the oxidation of the tire antidegradant 6PPD. 6PPD-Q is acutely toxic to select salmonids at environmentally relevant concentrations, while other fish species display tolerance to concentrations that surpass those measured in the environment. The reasons for these marked differences in sensitivity are presently unknown. The objective of this research was to explore potential toxicokinetic drivers of species sensitivity by characterizing biliary metabolites of 6PPD-Q in sensitive and tolerant fishes. For the first time, we identified an O-glucuronide metabolite of 6PPD-Q using high-resolution mass spectrometry. The semiquantified levels of this metabolite in tolerant species or life stages, including white sturgeon (Acipenser transmontanus), chinook salmon (Oncorhynchus tshawytscha), westslope cutthroat trout (Oncorhynchus clarkii lewisi), and nonfry life stages of Atlantic salmon (Salmo salar), were greater than those in sensitive species, including coho salmon (Oncorhynchus kisutch), brook trout (Salvelinus fontinalis), and rainbow trout (Oncorhynchus mykiss), suggesting that tolerant species might detoxify 6PPD-Q more effectively. Thus, we hypothesize that differences in species sensitivity are a result of differences in basal expression of biotransformation enzyme across various fish species. Moreover, the semiquantification of 6PPD-Q metabolites in bile extracted from wild-caught fish might be a useful biomarker of exposure to 6PPD-Q, thereby being valuable to environmental monitoring and risk assessment.

Growth regulation in brook charr Salvelinus fontinalis.

Fish growth can be modulated through genetic selection. However, it is not known whether growth regulatory mechanisms modulated by genetic selection can provide information about phenotypic growth variations among families or populations. Following a five-generation breeding program that selected for the absence of early sexual maturity and increased growth in brook charr we aimed to understand how the genetic selection process modifies the growth regulatory pathway of brook charr at the molecular level. To achieve this, we studied the regulation of growth traits at three different levels: 1) between lines-one under selection, the other not, 2) among-families expressing differences in average growth phenotypes, which we termed family performance, and 3) among individuals within families that expressed extreme growth phenotypes, which we termed slow- and fast-growing. At age 1+, individuals from four of the highest performing and four of the lowest performing families in terms of growth were sampled in both the control and selected lines. The gene expression levels of three reference and ten target genes were analyzed by real-time PCR. Results showed that better growth performance (in terms of weight and length at age) in the selected line was associated with an upregulation in the expression of genes involved in the growth hormone (GH)/insulin growth factor-1 (IGF-1) axis, including the igf-1 receptor in pituitary; the gh-1 receptor and igf-1 in liver; and ghr and igf-1r in white muscle. When looking at gene expression within families, family performance and individual phenotypes were associated with upregulations of the leptin receptor and neuropeptid Y-genes related to appetite regulation-in the slower-growing phenotypes. However, other genes related to appetite (ghrelin, somatostatin) or involved in muscle growth (myosin heavy chain, myogenin) were not differentially expressed. This study highlights how transcriptomics may improve our understanding of the roles of different key endocrine steps that regulate physiological performance. Large variations in growth still exist in the selected line, indicating that the full genetic selection potential has not been reached.

Molecular cloning and characterization of Cu-Zn superoxide dismutase (sod1) gene in brown trout and its expression in response to acute aquaculture stressors.

Aquaculture species are often exposed to acute stressors such as low water levels and handling during routine aquaculture procedures. This might result in oxidative stress by the increased reactive oxygen species (ROS)' production (e.g., superoxide anion). The harmful effects of ROS are eliminated by a defense system, referred antioxidant defense system (ADS). sod1 is the first gene involved in the ADS. Therefore, we cloned and characterized the open reading frame of the sod1 in brown trout. Then, we determined the effects of low water level and handling stress on sod1 mRNA expression in the liver and gills at 0 min, 1 and 2 h. The total RNA isolated was used to synthesize cDNA for RT-qPCR analysis. Phylogenetic tree, identity/similarity percentages, genomic organization, and conserved gene synteny analyses were applied to characterize Sod1/sod1. While low water level stress upregulated sod1 expression in the liver compared to the control group, no significant differences were observed in the gills between experimental groups. However, brown trout differently responded to handling stress at different time intervals in both tissues. Transcriptional differences were also noted between the sexes. This study contributes to the current understanding of the molecular mechanism between oxidative stress and ADS.

Proteomic analysis of the lake trout (Salvelinus namaycush) heart and blood: The beginning of a comprehensive lake trout protein database.

Lake trout (Salvelinus namaycush) are a top-predator species in the Laurentian Great Lakes that are often used as bioindicators of chemical stressors in the ecosystem. Although many studies are done using these fish to determine concentrations of stressors like legacy persistent, bioaccumulative and toxic chemicals, there are currently no proteomic studies on the biological effects these stressors have on the ecosystem. This lack of proteomic studies on Great Lakes lake trout is because there is currently no complete, comprehensive protein database for this species. Here, we employed proteomics approaches to develop a lake trout protein database that could aid in future research on this fish, in particular exposomics and adductomics. The current study utilized heart tissue and blood from two lake trout. Our previous work using lake trout liver revealed 4194 potential protein hits in the NCBI databases and 3811 potential protein hits in the UniProtKB databases. In the current study, using the NCBI databases we identified 838 proteins for the heart and 580 proteins for the blood tissues in the biological replicate 1 (BR1) and 1180 potential protein hits for the heart and 561 potential protein hits for the blood in BR2. Similar results were obtained using the UniProtKB databases. This study builds on our previous work by continuing to build the first comprehensive lake trout protein database and provides insight into protein homology through evolutionary relationships. This data is available via the PRIDE partner repository with the dataset identifier PXD023970.

Mercury source changes and food web shifts alter contamination signatures of predatory fish from Lake Michigan.

To understand the impact reduced mercury (Hg) loading and invasive species have had on methylmercury bioaccumulation in predator fish of Lake Michigan, we reconstructed bioaccumulation trends from a fish archive (1978 to 2012). By measuring fish Hg stable isotope ratios, we related temporal changes in Hg concentrations to varying Hg sources. Additionally, dietary tracers were necessary to identify food web influences. Through combined Hg, C, and N stable isotopic analyses, we were able to differentiate between a shift in Hg sources to fish and periods when energetic transitions (from dreissenid mussels) led to the assimilation of contrasting Hg pools (2000 to present). In the late 1980s, lake trout δ202Hg increased (0.4‰) from regulatory reductions in regional Hg emissions. After 2000, C and N isotopes ratios revealed altered food web pathways, resulting in a benthic energetic shift and changes to Hg bioaccumulation. Continued increases in δ202Hg indicate fish are responding to several United States mercury emission mitigation strategies that were initiated circa 1990 and continued through the 2011 promulgation of the Mercury and Air Toxics Standards rule. Unlike archives of sediments, this fish archive tracks Hg sources susceptible to bioaccumulation in Great Lakes fisheries. Analysis reveals that trends in fish Hg concentrations can be substantially affected by shifts in trophic structure and dietary preferences initiated by invasive species in the Great Lakes. This does not diminish the benefits of declining emissions over this period, as fish Hg concentrations would have been higher without these actions.

The effect of temperature on growth performance and aerobic metabolic scope in Arctic charr, Salvelinus alpinus (L.).

In recent years, Arctic char populations in Iceland have declined and the objective of this experiment was to throw further light on these changes by examining the effect of temperature (5, 9, 13, 17, and 21 °C) on the survival, growth rate, metabolism, and physiological indices of juvenile Arctic charr (initial mean body mass 4.02 ± 0.8 g). Mortality was 60% at 21 °C while at lower temperatures it was below 5%. However, Arctic charr populations in Iceland are declining in locations where the ambient temperature is lower, suggesting that other factors may be more important in determining the abundance of the species. The optimum temperature for growth was near 14 °C. The growth rate was progressively reduced at supra-optimum temperatures with almost no growth at 21 °C. Indicators of energy reserves: condition factor, relative intestinal mass, and hepatosomatic index are all consistent with reduced feed intake at supra-optimum temperatures. The standard and maximum metabolic rate (SMR; MMR), as well as the aerobic scope for activity (AS), were maximum at 13 °C. The routine metabolic rate (RMR) increased exponentially with temperature and, at T21, it was equal to the MMR suggesting, that the RMR was limited by the MMR. Moreover, increased heart- and gill mass at 21 °C are consistent with increased stress on the cardiovascular system. These findings are in keeping with the OCLTT hypothesis that the thermal tolerance of fish is limited by the capacity of the cardiovascular system to deliver oxygen and support metabolism. Taken together, the results of this experiment suggest, that growth rate is reduced at supra-optimum temperatures because of reduced energy intake, increased metabolic demand, and limitations in the capacity of the cardiovascular system to support metabolic rate at high temperatures. At lower temperatures, growth does not appear to be limited by the AS.

Evaluation of Proteomic and Lipidomic Changes in Aeromonas-Infected Trout Kidney Tissue with the Use of FT-IR Spectroscopy and MALDI Mass Spectrometry Imaging.

Aeromonas species are opportunistic bacteria causing a vast spectrum of human diseases, including skin and soft tissue infections, meningitis, endocarditis, peritonitis, gastroenteritis, and finally hemorrhagic septicemia. The aim of our research was to indicate the molecular alterations in proteins and lipids profiles resulting from Aeromonas sobria and A. salmonicida subsp. salmonicida infection in trout kidney tissue samples. We successfully applied FT-IR (Fourier transform infrared) spectroscopy and MALDI-MSI (matrix-assisted laser desorption/ionization mass spectrometry imaging) to monitor changes in the structure and compositions of lipids, secondary conformation of proteins, and provide useful information concerning disease progression. Our findings indicate that the following spectral bands' absorbance ratios (spectral biomarkers) can be used to discriminate healthy tissue from pathologically altered tissue, for example, lipids (CH2/CH3), amide I/amide II, amide I/CH2 and amide I/CH3. Spectral data obtained from 10 single measurements of each specimen indicate numerous abnormalities concerning proteins, lipids, and phospholipids induced by Aeromonas infection, suggesting significant disruption of the cell membranes. Moreover, the increase in the content of lysolipids such as lysophosphosphatidylcholine was observed. The results of this study suggest the application of both methods MALDI-MSI and FT-IR as accurate methods for profiling biomolecules and identifying biochemical changes in kidney tissue during the progression of Aeromonas infection.

Health benefits and risks associated with element uptake from grilled fish and fish products.

BACKGROUND: The content of essential and toxic elements in grilled fish (Atlantic salmon, trout) and in 20 assortments of fish products was examined. The aim of this study was to assess the fulfilment of the demand for Zn, Fe, Mn and Cu based on recommended dietary allowances (RDAs) or adequate intakes (AIs) and to assess the health risk associated with the consumption of Al, Pb and Cd. The risk assessment was based on estimated weekly intake (EWI), hazard index (HI), target risk ratio (THQ) and percentage: provisional tolerable weekly intake (PTWI) for Al, tolerable weekly intake (TWI) for Cd and reference dose lower bound (BMDL) for Pb. RESULTS: Taking into account the health benefits, the best source of Fe, Cu and Mn was found to be fish products in cans and jars, 150 g of which covered the daily requirement: Fe 9.39%, Cu 2.91% and Mn 1.21%. Smoked fish covered the RDA for Zn to 5.69%. Moreover, the uptake of toxic elements was as follows: Al 0.45% PTWI: Pb 0.74% BMDL10 and Cd 2.20% TWI. The THQ values for Pb and Cd were significantly lower than 1, whereas for Al it reached as high as 4.0. CONCLUSION: The obtained results indicate that there is no risk related to the intake of Pb and Cd with the consumption of the tested fish products. Low consumption of fish by Poles, however, contributes to their small share in covering the demand for microelements, i.e. Zn, Fe, Mn and Cu. © 2021 Society of Chemical Industry.

Vitellogenin Induction in Mucus from Brook Trout (Salvelinus fontinalis).

Induction of vitellogenin (VTG) is widely used as a biomarker of exposure of male or immature fish to chemicals that are agonists of the estrogen receptor (i.e., xenoestrogens). Analysis of VTG in samples of epidermal mucosa collected from fish is a non-invasive method for evaluating whether wild fish are exposed to xenoestrogens. In this study, the mean levels of VTG in the mucus of immature brook trout (Salvelinus fontinalis) collected from the Credit River in Ontario, Canada downstream of aging residential septic systems and in an agricultural watershed were 0.67 ng per mg protein, which was significantly elevated relative to the mean VTG levels of 0.22 ng per mg protein in the mucus of immature brook trout collected from a less impacted site. To validate the mucus assay, immature brook trout were exposed in the laboratory to 17α-ethinylestradiol (EE2) at nominal concentrations of 10, 50 and 100 ng/L and VTG levels in mucus from these fish showed a concentration-dependent increase relative to fish from the control treatment. This study illustrates the utility of this non-lethal method for assessing whether wild fish have been exposed in situ to xenoestrogens. Exposures to xenoestrogens from non-point sources may be impacting brook trout populations in urban watersheds in southern Ontario.

How do exercise training variables stimulate processes related to mitochondrial biogenesis in slow and fast trout muscle fibres?

NEW FINDINGS: What is the central question of this study? Exercise is known to promote mitochondrial biogenesis in skeletal muscle, but what are the most relevant training protocols to stimulate it? What is the main finding and its importance? As in mammals, training in rainbow trout affects slow and fast muscle fibres differently. Exercise intensity, relative to volume, duration and frequency, is the most relevant training variable to stimulate the processes related to mitochondrial biogenesis in both red and white muscles. This study offers new insights into muscle fibre type-specific transcription and expression of genes involved in mitochondrial adaptations following training. ABSTRACT: Exercise is known to be a powerful way to improve health through the stimulation of mitochondrial biogenesis in skeletal muscle, which undergoes cellular and molecular adaptations. One of the current challenges in human is to define the optimal training stimulus to improve muscle performance. Fish are relevant models for exercise training physiology studies mainly because of their distinct slow and fast muscle fibres. Using rainbow trout, we investigated the effects of six different training protocols defined by manipulating specific training variables (such as exercise intensity, volume, duration and frequency), on mRNAs and some proteins related to four subsystems (AMP-activated protein kinase-peroxisome proliferator-activated receptor γ coactivator-1α signalling pathway, mitochondrial function, antioxidant defences and lactate dehydrogenase (LDH) metabolism) in both red and white muscles (RM and WM, respectively). In both muscles, high-intensity exercise stimulated more mRNA types and enzymatic activities related to mitochondrial biogenesis than moderate-intensity exercise. For volume, duration and frequency variables, we demonstrated fibre type-specific responses. Indeed, for high-intensity interval training, RM transcript levels are increased by a low training volume, but WM transcript responses are stimulated by a high training volume. Moreover, transcripts and enzymatic activities related to mitochondria and LDH show that WM tends to develop aerobic metabolism with a high training volume. For transcript stimulation, WM requires a greater duration and frequency of exercise than RM, whereas protein adaptations are efficient with a long training duration and a high frequency in both muscles.

Life through a wider scope: Brook Trout (Salvelinus fontinalis) exhibit similar aerobic scope across a broad temperature range.

Brook Trout (Salvelinus fontinalis) have been widely introduced throughout the world and are often considered as direct competitors with native salmonid species. Metabolic rate is one metric we can examine to improve our understanding of how well fish perform in different habitats, including across temperature gradients, as metabolism can be directly influenced by environmental temperatures in ectotherms. We estimated the standard metabolic rate, maximum metabolic rate, and aerobic scope of lab-reared juvenile Brook Trout (~1 year) using intermittent-flow respirometry across a range of temperatures (5-23 °C) likely experienced in the wild. We included a diurnal temperature cycle of ±1.5 °C for each treatment temperature to simulate temporal variation observed in natural waterbodies. Standard metabolic rate and maximum metabolic rate both increased with acclimation temperature before appearing to plateau around 20 °C, while mass specific aerobic scope was found to increase from a mean of 287.25 ± 13.03 mg O2·kg-1·h-1 at 5 °C to 384.85 ± 13.31 mg O2·kg-1·h-1 at 15 °C before dropping at higher temperatures. Although a slight peak was found at 15 °C, the generally flat thermal performance curve for aerobic scope suggests Brook Trout are capable of adjusting to a relatively wide range of thermal regimes, appearing to be eurythermal, or a thermal generalist, at least for salmonids. The ability of this population to maintain similar physiological performance across a wide range of temperatures may help explain why Brook Trout succeed in a variety of different thermal habitats.

Development of a capillary electrophoresis-mass spectrometry method for the analysis of metformin and its transformation product guanylurea in biota.

A method with capillary electrophoresis coupled to mass spectrometry was optimized to determine the uptake of metformin and its metabolite guanylurea by zebrafish (Danio rerio) embryos and brown trout (Salmo trutta f. fario) exposed under laboratory conditions. Metformin was extracted from fish tissues by sonication in methanol, resulting in an absolute recovery of almost 90%. For the extraction of guanylurea from brown trout, solid-phase extraction was implemented with a recovery of 84%. The use of a mixture of methanol and glacial acetic acid as a non-aqueous background electrolyte was vital to achieve robust analysis using a bare fused-silica capillary with an applied voltage of +30 kV. Problems with adsorption associated with an aqueous background electrolyte were eliminated using a non-aqueous background electrolyte made of methanol/acetic acid (97:3) with 25 mM ammonium acetate (for zebrafish embryos) or 100 mM ammonium acetate (for brown trouts), depending on the sample complexity and matrix influences. High resolution and high separation selectivity from matrix components were achieved by optimization of the ammonium acetate concentration in the background electrolyte. An extensive evaluation of matrix effects was conducted with regard to the complex matrices present in the fish samples. They required adapting the background electrolyte to higher concentrations. Applying this method to extracts of zebrafish embryos and brown trout tissue samples, limits of detection for both metformin and guanylurea in zebrafish embryos (12.2 µg/l and 15 µg/l) and brown trout tissues (15 ng/g and 34 ng/g) were in the low µg/l or ng/g range. Finally, metformin and guanylurea could be both quantified for the first time in biota samples from exposure experiments.

The effects of the neurotoxic agent emamectin benzoate on the expression of immune and stress-related genes and blood serum profiles in the Rainbow trout.

Emamectin, a neurotoxic agent, is a semi-synthetic insecticide that belongs to the Avermectin family and is used against helmintic infections in the Salmonidae family. Its secondary effects are not clear; thus, the aim of this study was to investigate the only effects of emamectin benzoate on various biochemical parameters (AST, ALT, GGT, total protein, albumin and glucose) in serum and expressional changes of IL-1ß, TNF-α, HSP70 and IL-8 in liver and spleen. For the purpose stated above, rainbow trout (n = 15) were administered 50 µg EB per kg fish daily for 7, 14 and 21 days. The results indicated that weight gains did not change (p > 0.05), AST increased at day 21 (p < 0.05), while the changes of other biochemical parameters were not significant (p > 0.05). The changes in expression of IL-1ß, TNF-α and HSP70 were significant (p < 0.05), while the changes of IL-8 expressions were not significant (p ˃ 0.05). In a conclusion, EB changed immun and stress-related gene expression in liver and spleen, and furthermore, AST changed in a dose- and time-dependent manner. The results imply that emamectin benzoate cause stress. This study is helpful to understand the effects of avermectin pharmaceutical family.

Behavioral responses to annual temperature variation alter the dominant energy pathway, growth, and condition of a cold-water predator.

There is a pressing need to understand how ecosystems will respond to climate change. To date, no long-term empirical studies have confirmed that fish populations exhibit adaptive foraging behavior in response to temperature variation and the potential implications this has on fitness. Here, we use an unparalleled 11-y acoustic telemetry, stable isotope, and mark-recapture dataset to test if a population of lake trout (Salvelinus namaycush), a cold-water stenotherm, adjusted its use of habitat and energy sources in response to annual variations in lake temperatures during the open-water season and how these changes translated to the growth and condition of individual fish. We found that climate influenced access to littoral regions in spring (data from telemetry), which in turn influenced energy acquisition (data from isotopes), and growth (mark-recapture data). In more stressful years, those with shorter springs and longer summers, lake trout had reduced access to littoral habitat and assimilated less littoral energy, resulting in reduced growth and condition. Annual variation in prey abundance influenced lake trout foraging tactics (i.e., the balance of the number and duration of forays) but not the overall time spent in littoral regions. Lake trout greatly reduced their use of littoral habitat and occupied deep pelagic waters during the summer. Together, our results provide clear evidence that climate-mediated behavior can influence the dominant energy pathways of top predators, with implications ranging from individual fitness to food web stability.

A seasonal comparison of trace metal concentrations in the tissues of Arctic charr (Salvelinus alpinus) in Northern Québec, Canada.

Ecotoxicological research detailing trace metal contamination and seasonal variation in the tissues of northern fishes such as Arctic charr (Salvelinus alpinus) has been poorly represented in the literature beyond examination of mercury. In an effort to address this, anadromous Arctic charr were collected from the Deception River watershed in the late summer and post-winter season, before quantifying seasonal and organotropic variations in dorsal muscle and liver concentrations of arsenic, cadmium, chromium, copper, nickel, lead, and zinc. Potential linkages with biological variables (fork length, age, and somatic condition) and indicators of feeding behavior (δ13C and δ15N) were also assessed. Trace metal organotropism favouring elevation in liver tissue concentrations was exhibited by cadmium, copper, nickel and zinc, while arsenic, chromium and lead exhibited no significant organotropic variation. Seasonal differences in concentrations were metal and tissue dependent, but generally increased in tissues collected from post-winter sampled Arctic charr. Significant correlations with biological and trophic descriptors were also determined to be element and tissue dependent. These parameters, in addition to season, were incorporated into multi-predictor variable models, where variations in trace metal concentration data were often best explained when season, somatic condition, and trophic descriptors were included. These variables were also of greatest relative importance across all considered trace metals and tissue types. These findings suggest that seasonally linked processes have the greatest influence on trace metal concentrations in anadromous Arctic charr. Future metal-related research on Arctic charr and other northern fish species should further consider these variables when evaluating elemental accumulation.

Identification and Expression Profiling of Toll-Like Receptors of Brown Trout (Salmo trutta) during Proliferative Kidney Disease.

Proliferative kidney disease is an emerging disease among salmonids in Europe and North America caused by the myxozoan parasite Tetracapsuloides bryosalmonae. The decline of endemic brown trout (Salmo trutta) in the Alpine streams of Europe is fostered by T. bryosalmonae infection. Toll-like receptors (TLRs) are a family of pattern recognition receptors that acts as sentinels of the immune system against the invading pathogens. However, little is known about the TLRs' response in salmonids against the myxozoan infection. In the present study, we identified and evaluated TLR1, TLR19, and TLR13-like genes of brown trout using data-mining and phylogenetic analysis. The expression pattern of TLRs was examined in the posterior kidney of brown trout infected with T. bryosalmonae at various time points. Typical Toll/interleukin-1 receptor protein domain was found in all tested TLRs. However, TLR13-like chr2 had a short amino acid sequence with no LRR domain. Phylogenetic analysis illustrated that TLR orthologs are conserved across vertebrates. Similarly, a conserved synteny gene block arrangement was observed in the case of TLR1 and TLR19 across fish species. Interestingly, all tested TLRs showed their maximal relative expression from 6 to 10 weeks post-exposure to the parasite. Our results suggest that these TLRs may play an important role in the innate defense mechanism of brown trout against the invading T. bryosalmonae.

Genistein Induces Adipogenic and Autophagic Effects in Rainbow Trout (Oncorhynchus mykiss) Adipose Tissue: In Vitro and In Vivo Models.

Soybeans are one of the most used alternative dietary ingredients in aquafeeds. However, they contain phytoestrogens like genistein (GE), which can have an impact on fish metabolism and health. This study aimed to investigate the in vitro and in vivo effects of GE on lipid metabolism, apoptosis, and autophagy in rainbow trout (Oncorhynchus mykiss). Primary cultured preadipocytes were incubated with GE at different concentrations, 10 or 100 µM, and 1 µM 17ß-estradiol (E2). Furthermore, juveniles received an intraperitoneal injection of GE at 5 or 50 µg/g body weight, or E2 at 5 µg/g. In vitro, GE 100 µM increased lipid accumulation and reduced cell viability, apparently involving an autophagic process, indicated by the higher LC3-II protein levels, and higher lc3b and cathepsin d transcript levels achieved after GE 10 µM. In vivo, GE 50 µg/g upregulated the gene expression of fatty acid synthase (fas) and glyceraldehyde-3-phosphate dehydrogenase in adipose tissue, suggesting enhanced lipogenesis, whereas it increased hormone-sensitive lipase in liver, indicating a lipolytic response. Besides, autophagy-related genes increased in the tissues analyzed mainly after GE 50 µg/g treatment. Overall, these findings suggest that an elevated GE administration could lead to impaired adipocyte viability and lipid metabolism dysregulation in rainbow trout.

Impact of Endocrine Disruptors on Vitellogenin Concentrations in Wild Brown Trout (Salmo trutta trutta).

The adverse effects of endocrine disruptors (EDs) on aquatic wildlife and human health represent a current issue of high public concern. Substantial knowledge of the level of estrogenic EDs in fish has accumulated from field surveys. For this purpose, a survey of wild brown trout (Salmo trutta trutta) was carried out to assess the incidence of EDs in the feral fish population living in the Liri river (Abruzzi, Italy). The results of this study show that this aquatic environment possesses an estrogenic potency that triggered the increase of vitellogenin levels in both female and male trouts. Fish exposed to different pesticides and urban waste in downstream river showed higher vitellogenin levels in comparison to the headwater site. Furthermore, some trouts coming from the downstream reported the presence of several pesticides and fungicides, some of these banned several years ago.