Eubiotic Effect of a Dietary Bio-Aqua® and Sodium Diformate (NaDF) on Salmo trutta caspius: Innate Immune System, Biochemical Indices, Antioxidant Defense, and Expression of Immunological and Growth-Related Genes.

The present study investigated the effects of combined and singular oral administration of Bio-Aqua® with different dosages of sodium diformate (NaDF) on biochemical indices, innate immune responses, antioxidant effects, and expressions of immunological related genes of Caspian brown trout (Salmo trutta caspius). Fingerlings Salmo trutta caspius (n = 1800; initial weight 15 ± 3 g) were randomly allocated into five groups (120 fish group-1 in triplicates). Control diet: without any addition, G1, G2, G3, and G4 received diets containing 0.2 g kg-1 commercial probiotic Bio-Aqua® combined with 0, 0.5, 1.0, and 1.5% NaDF to the basal diet for 60 days according to recommended dosages reported in previous studies. Results indicated that serum bactericidal activity (G3 on day 60 and G1 on day 30) and classic complement in all groups (on day 60) (G1 and G2 on day 30) were significantly elevated (P < 0.05). The serum lysozyme, glucose, globulin, and albumin levels showed no significant differences between all groups compared to the control group (P > 0.05). On days 30 and 60 of the sampling, no significant difference was observed in the amount of superoxide disotase (SOD) and catalase (CAT) between the treatments (P > 0.05) but activity of malondialdehyde (MDA) was lower in G1 than the control (P < 0.05). The expression of the immune-regulating genes IL-10, IL-1ß, GTP, FATP, and IGF was significantly improved in all probiotic + acidifier-treated groups (P < 0.05). The current findings showed that mixture of Bio-Aqua® and NaDF (1.5% + pro) is beneficial, as it effectively improves some immune parameters and expression of immunological and growth-related genes in Caspian brown trout.

Effect of Astaxanthin on the Antioxidant Capacity and Intestinal Microbiota of Tsinling Lenok Trout (Brachymystax lenok tsinlingensis).

Astaxanthin (Ast) has been shown to be beneficial for the antioxidant capacity, immune system, and stress tolerance of fish. This study was conducted to investigate the effects of dietary supplementation of Ast on the antioxidant capacity and intestinal microbiota of tsinling lenok trout. We formulated four diets with 0 (CT), 50 (A50), 100 (A100), and 150 (A150) mg/kg Ast. The results showed that Ast increased total superoxide dismutase (T-SOD) and glutathione peroxidase (GSH-Px), lysozyme (LZM), and catalase (CAT) activities. Malondialdehyde (MDA) content was lower in A150 and A100 than in CT (P < 0.05). Furthermore, the activities of acid phosphatase (ACP) were higher in A100 and A150 than in CT (P < 0.05). We harvested the midgut and applied next-generation sequencing of 16S rDNA. Compared to the control group, the Ast group had a greater abundance of Halomonas. Functional analysis showed that polycyclic aromatic hydrocarbon degradation was significantly higher with Ast, while novobiocin biosynthesis and C5-branched dibasic acid metabolism were significantly lower with Ast. In conclusion, Ast could enhance the antioxidant capacity, non-specific immunity, and intestinal health of tsinling lenok trout.

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.

Influence of dietary linseed oil as substitution of fish oil on whole fish fatty acid composition, lipid metabolism and oxidative status of juvenile Manchurian trout, Brachymystax lenok.

In this study, juvenile Manchurian trout, Brachymystax lenok (initial weight: 6.43 ± 0.02 g, mean ± SE) were received for nine weeks with five types of diets prepared by gradually replacing the proportion of fish oil (FO) with linseed oil (LO) from 0% (LO0) to 25% (LO25), 50% (LO50), 75% (LO75), and 100% (LO100). The eicosapentaenoic (EPA) and docosahexaenoic (DHA) composition decreased with increasing inclusion level of LO (P < 0.05). With increasing LO inclusion level, triglyceride (TAG) content of serum increased significantly, however, there was a decrease in high-density lipoprotein cholesterol (HDL) (P < 0.05). LO substitution of FO up-regulated the gene expression level of lipid metabolism-related genes Fatty Acid Desaturases 6 (FAD6), Acetyl-Coa Carboxylase (ACCα), Sterol Regulatory Element Binding Protein 1 (SREBP-1), and Sterol O- Acyl Transferase 2 (SOAT2), and down-regulated the gene expression level of Peroxisome Proliferator-Activated Receptor a (PPARα) (P < 0.05). The SOD activities of both serum and liver in LO100 were significantly lower than in LO25 (P < 0.05). The CAT activity of the liver in LO100 was significantly lower than in LO0 and LO25 (P < 0.05). This study indicates that the Manchurian trout may have the ability to synthesize LC-PUFAs from ALA, and an appropriate LO in substitution of FO (<75%) could improve both the lipid metabolism and the oxidation resistance.

Early Evidence for Northern Salmonid Fisheries Discovered using Novel Mineral Proxies.

Salmonid resources currently foster socioeconomic prosperity in several nations, yet their importance to many ancient circumpolar societies is poorly understood due to insufficient fish bone preservation at archaeological sites. As a result, there are serious gaps in our knowledge concerning the antiquity of northern salmonid fisheries and their impacts on shaping biodiversity, hunter-gatherer adaptations, and human-ecological networks. The interdisciplinary study presented here demonstrates that calcium-magnesium phosphate minerals formed in burned salmonid bones can preserve at ancient northern sites, thus informing on the early utilization of these resources despite the absence of morphologically classifiable bones. The minerals whitlockite and beta magnesium tricalcium phosphate were identified in rare morphologically classifiable Atlantic salmonid bones from three Mid-Holocene sites in Finland. Large amounts of beta magnesium tricalcium phosphate were also experimentally formed by burning modern Atlantic salmonid and brown trout bones. Our results demonstrate the value of these minerals as proxies for ancient northern salmonid fishing. Specifically, the whitlockite mineral was discovered in hearth sediments from the 5,600 year old Yli-Ii Kierikinkangas site on the Iijoki River in northern Finland. Our fine sieving and mineralogical analyses of these sediments, along with zooarchaeological identification of recovered bone fragments, have confirmed for the first time that the people living at this village did incorporate salmonids into their economies, thus providing new evidence for early estuary/riverine fisheries in northern Finland.

Oleaginous yeast as a component in fish feed.

This study investigates the replacement of vegetable oil (VO) in aquaculture feed for Arctic char (Salvelinus alpinus) with oil produced by the oleaginous yeast Lipomyces starkeyi grown in lignocellulose (wheat straw) hydrolysate. VO is extensively used to partially replace fish oil in aquaculture feed, which can be seen as non-sustainable. VO itself is becoming a limited resource. Plant oils are used in many different applications, including food, feed and biodiesel. Its replacement in non-food applications is desirable. For this purpose, yeast cells containing 43% lipids per g dry weight were mechanically disrupted and incorporated into the fish feed. There were no significant differences in this pilot study, regarding weight and length gain, feed conversion ratio, specific growth rate, condition factor and hepatosomatic index between the control and the yeast oil fed group. Fatty and amino acid composition of diet from both groups was comparable. Our results in fish demonstrate that it is possible to replace VO by yeast oil produced from lignocellulose, which may broaden the range of raw materials for food production and add value to residual products of agriculture and forestry.

Feedback from Arctic charr: Feed flavour stimulation and re-feeding after feed deprivation stimulate genes encoding both orexigenic and anorexigenic neuropeptides.

Despite vast research attention, the knowledge about central mechanisms of appetite regulation in teleost remains inconclusive. A common strategy in studies on appetite regulating mechanisms is to measure the response to feed restriction or - deprivation, but responses vary between fish species and between experiments, and are also likely dependent on the degree of energy perturbation. The anadromous Arctic charr is an interesting model for studying appetite regulation as its feeding cycle comprises months of winter anorexia, and hyperphagia during summer. Here we studied how the gene expression of putative hypothalamic appetite regulators were affected by two days, one week and one month feed deprivation during summer, and subsequent re-feeding and exposure to feed flavour. Short-term feed deprivation caused only a minor reduction in condition factor and had no effect on hypothalamic gene expression. Long-term feed-deprivation caused a marked reduction in weight and condition factor which contrasted the increase in weight and condition factor seen in ad libitum fed controls. A marked energy perturbation by feed deprivation was also indicated by a lower hypothalamic expression of the genes encoding insulin-like growth factor 1 (IGF1) and IGF1 binding protein 5 in the feed deprived charr compared to fed controls. Surprisingly, long-term feed deprivation and energy perturbation did not induce changes in hypothalamic appetite regulators. Unexpectedly, re-feeding and exposure to feed flavour caused an increase in the expression of the genes encoding the orexigenic agouti-related peptide and the anorexigenic melanocortin receptor 4 and cocaine- and amphetamine-regulated transcript. Our study gives strong evidence for a role of these in appetite regulation in Arctic charr, but their mechanisms of action remain unknown. We suggest that changes in gene expression are more likely to be registered during transition phases, e.g. from fasting to feeding and upon stimulatory inputs such as feed flavour.

Differential Accumulation of Mercury and Selenium in Brown Trout Tissues of a High-Gradient Urbanized Stream in Colorado, USA.

Total mercury (THg) and selenium (Se) were analyzed by Inductively Coupled Plasma Mass Spectrometry in 11 internal and external tissues and stomach contents from 23 brown trout, Salmo trutta, of a 22.9-km reach of a high-gradient stream (upper Fountain Creek) in Colorado, USA, impacted by coal-fired power plants, shale deposits, and urbanization. Trout and water were sampled from four sites ranging from 2335 to 1818 m elevation. Lengths, weights, and ages of fish between pairs of the four sites were not significantly different. The dry weight (dw) to wet weight (ww) conversion factor for each tissue was calculated with egg-ovary highest at 0.379 and epaxial muscle fourth highest at 0.223. THg and Se in stomach contents indicated diet and not ambient water was the major source of Hg and Se bioaccumulated. Mean THg ww in kidney was 40.33 µg/kg, and epaxial muscle second highest at 36.76 µg/kg. None of the tissues exceeded the human critical threshold for Hg. However, all 23 trout had at least one tissue type that exceeded 0.02 mg/kg THg ww for birds, and four trout tissues exceeded 0.1 mg/kg THg ww for mammals, indicating that piscivorous mammals and birds should be monitored. Se concentrations in tissues varied depending on ww or dw listing. Mean Se dw in liver was higher than ovary at the uppermost site and the two lower sites. Liver tissue, in addition to egg-ovary, should be utilized as an indicator tissue for Se toxicity.

Dietary peppermint (Mentha piperita) extracts promote growth performance and increase the main humoral immune parameters (both at mucosal and systemic level) of Caspian brown trout (Salmo trutta caspius Kessler, 1877).

The effects of dietary administration of peppermint (Mentha piperita L.) on Caspian brown trout fish (Salmo trutta caspius) were studied. Fish were divided into 4 groups before being fed diets supplemented with 0% (control), 1%, 2% and 3% of peppermint extracts for 8 weeks. Dose-dependent increases in growth, immune (both in skin mucus and blood serum) and hematological parameters (number of white cells, hematocrit and hemoglobin content), as well as in amylase activity and in the number of lactic acid bacteria on intestine were recorded in fish fed supplemented diets compared to control fish. However, the dietary peppermint supplements have different effects on the number of blood leucocytes depending on the leukocyte cell type. While no significant differences were observed in the number of blood monocytes and eosinophils, the number of lymphocytes was decreased, respectively, on fish fed peppermint enriched diets, respect to the values found in control fish. Furthermore, dietary peppermint supplements have no significant effect on blood biochemical parameters, enzymatic activities of liver determined in serum and total viable aerobic bacterial count on intestine of Caspian brown trout. Present results support that dietary administration of peppermint promotes growth performance and increases the main humoral immune parameters (both at mucosal and systemic level) and the number of the endogenous lactic acid bacteria of Caspian brown trout. This study underlying several positive effects of dietary administration of peppermint to farmed fish.

Intestinal nematodes affect selenium bioaccumulation, oxidative stress biomarkers, and health parameters in juvenile rainbow trout (Oncorhynchus mykiss).

In environmental studies, parasites are often seen as a product of enhanced host susceptibility due to exposure to one or several stressors, whereas potential consequences of infections on host responses are often overlooked. Therefore, the present study focused on effects of parasitism on bioaccumulation of selenium (Se) in rainbow trout (Oncorhynchus mykiss). Joint effects of biological (parasite) and chemical (Se) stressors on biomarkers of oxidative stress (glutathione-S-transferase (GST), superoxide dismutase (SOD)), and fish health (condition factor (K), hepatosomatic index (HSI), gross energy) were also examined. Fish of the control group received uncontaminated food, while test fish, either experimentally infected with the nematode Raphidascaris acus or not, were exposed to dietary selenomethionine (Se-Met) at an environmentally relevant dose over 7 weeks. Selenium bioaccumulation by the parasite was low relative to its host, and parasitized trout showed slowed Se accumulation in the muscle as compared to uninfected fish. Furthermore, GST and SOD activities of trout exposed to both Se-Met and parasites were generally significantly lower than in fish exposed to Se-Met alone. Gross energy concentrations, but not K or HSI, were reduced in fish exposed to both Se-Met and R. acus. Together the experiment strongly calls for consideration of parasites when interpreting effects of pollutants on aquatic organisms in field investigations.

Selenium bioaccumulation in stocked fish as an indicator of fishery potential in pit lakes on reclaimed coal mines in Alberta, Canada.

Pit lakes are a common reclamation strategy for open pit mines; however, there is a concern about their water quality and suitability as fish habitat because they are often contaminated by metals or metalloids. This study assessed the exposure of fish and invertebrates to selenium (Se) and other metals and metalloids in pit lakes formed by open pit coal mining in Tertiary (thermal coal) and in Cretaceous (metallurgical coal) bedrock. Juvenile hatchery rainbow trout, Oncorhynchus mykiss, and brook trout, Salvelinus fontinalis, were stocked into two thermal coal pit lakes (water Se < 2 µg/L, low water Se) and two metallurgical coal pit lakes (water Se > 15 µg/L, high water Se). Se accumulation in stocked fish and concentrations in invertebrates were characterized over a period of 2 years. In the metallurgical pits, invertebrates had higher Se concentrations and fish accumulated Se to higher levels (exceeding USEPA tissue Se guidelines) than biota in the thermal pits. Rainbow and brook trout accumulated similar concentrations of Se in their muscle and exhibited a similar relationship between whole-body and muscle Se concentrations. These results may be used by resource managers to assess compliance with whole-body tissue Se guidelines and to determine if pit lakes in coal mining areas pose a significant Se risk to wildlife or human health. The high Se exposure in metallurgical coal pits indicates that under the current mining and reclamation strategy, these lakes are not suitable for management as recreational "put and take" fisheries.

Selenium species and their distribution in freshwater fish from Argentina.

The distribution and speciation of selenium (Se) in freshwater fish (muscle and liver tissue) from lakes in Argentina was investigated. Three introduced species, brown trout (Salmo trutta), rainbow trout (Oncorhynchus mykiss) and brook trout (Salvelinus fontinalis), and one native species, creole perch (Percichthys trucha), were investigated. Values for total selenium in muscle ranged from 0.66 to 1.61 µg/g, while in the liver, concentrations were much higher, from 4.46 to 73.71 µg/g on a dry matter basis. Separation of soluble Se species (SeCys(2), selenomethionine (SeMet), SeMeSeCys, selenite and selenate) was achieved by ion exchange chromatography and detection was performed by inductively coupled plasma-mass spectrometry. The results showed that in fish muscle, from 47 to 55 % of selenium was soluble and the only Se species identified was SeMet, which represented around 80 % of soluble Se, while in the liver, the amount of soluble Se ranged from 61 to 76 % and the percentage of species identified (SeMet and SeCys(2)) was much lower and ranged from 8 to 17 % of soluble Se.

Determination of bioaccumulation of heavy metals and selenium in tissues of brown trout Salmo trutta macrostigma (Duméril, 1858) from Munzur Stream, Tunceli, Turkey.

The objective of the present work was to determine the bioaccumulation of arsenic (As), cadmium (Cd), copper (Cu), lead (Pb), mercury (Hg), uranium (U) and selenium (Se) in gill, liver, and muscle tissues of the fresh water fish Salmo trutta macrostigma (Duméril, 1858) in Munzur Stream, Tunceli, Turkey. The highest concentrations of U (1.83 µg kg(-1)), Pb (119.84 µg kg(-1)) and Se (1.31 µg kg(-1)) were recorded in the gills of S. t. macrostigma. Concentrations of As (46.27 µg kg(-1)), Cd (109.19 µg kg(-1)), Hg (16.40 µg kg(-1)), Cu (18.19 µg kg(-1)) were recorded at highest levels in the liver. The results showed that there were significant differences in concentrations of As, Cd, Cu, Pb, Se, U and Hg in gill, liver and muscle tissue (p < 0.05). Heavy metals were within the edible parts of the investigated fish were in the permissible safety levels for human uses.

Effects of dietary vitamin B1 (thiamine) and magnesium on the survival, growth and histological indicators in lake trout (Salvelinus namaycush) juveniles.

An interaction of two essential nutrients, thiamine and magnesium (Mg) has been documented in in vitro and in vivo studies in mammalian metabolism. However, the role of this association in poikilothermic vertebrates, such as fish, remains elusive. The purpose of this study was first to investigate the effects of dietary thiamine and Mg, and their interaction in lake trout and second to better understand the mechanism leading to early mortality syndrome (EMS), which is caused by a low thiamine level in embryos of many species of salmonids in the wild. Semi-purified diets (SPD) were prepared to accomplish 2 × 2 factorial design that were either devoid of or supplemented with thiamine mononitrate (20 mg/kg diet), magnesium oxide (700 mg/kg diet), or both. Lake trout alevins at the swim-up stage were fed for 10 wk one of the SPD diets or a commercial diet at the same rate (2.0-1.5%) based on recorded biomass. Our results showed that the concentrations of thiamine in the trunk muscle and Mg of whole body were closely associated with the dietary level of two nutrients. The interaction of low dietary Mg and thiamine resulted in apparently worsened overt symptoms of thiamine deficiency in lake trout leading to a higher mortality of fish during the seven week long trial (P<0.05). The fish fed a thiamine-devoid and Mg-supplemented diet were presumed to survive longer (10 wk) than the fish fed diets devoid of both nutrients (discontinued after 7th wk due to high mortality). However, we did not observe histopathological changes in the brain and liver corresponding to thiamine concentrations in tissues. These data suggest that Mg enhanced utilization of the thiamine remaining in the fish body and its interdependence was consistent with observations in mammals. EMS severity might be worsened when Mg is deficient in parental diets (and consequently in yolk sac) and/or first feed.

Selenium moderates mercury toxicity in free-ranging freshwater fish.

Due to the extremely high affinity of selenium (Se) to mercury (Hg), Se sequesters Hg and reduces its biological availability in organisms. However the converse is also true. Hg sequesters Se, causing Hg to inhibit the formation of Se dependent enzymes while supplemental Se supports their continued synthesis. Hence, whether or not toxic effects accompany exposure to Hg depends upon the tissue Se:Hg molar ratio of the organism. The main objective of the present study was to investigate how levels of Hg and Se affected metallothionein (MT) induction in free-ranging brown trout, Salmo trutta, from Lake Mjøsa, Norway (a Se depauperate lake). MT is proposed as a sensitive biomarker of potential detrimental effects induced by metals such as Hg. Emphasis was addressed to elucidate if increased tissue Se:Hg molar ratios and Se levels affected the demands for MT in the trout. The Se:Hg molar ratio followed by tissue Se levels were most successful for assessing the relationship between metal exposure and MT levels in the trout. Thus, Hg in molar excess over Se was a stronger inducer of MT synthesis than tissue Hg levels in the trout, supporting the assumption that Se has a prominent protective effect against Hg toxicity. Measuring Hg in animals may therefore provide an inadequate reflection of the potential health risks to humans and wildlife if the protective effects of Se are not considered.

Low concentrations of selenium in stream food webs of eastern Canada.

Herbivorous and predatory invertebrates and two species of fish (brook trout and blacknose dace) were collected from 49 streams in New Brunswick, Canada to determine whether concentrations of selenium (Se) in the biota were affected by a point source (a coal-fired power plant), and stream water chemistry (pH, sulphate, conductivity, and total organic carbon), and to determine the trophic transfer of Se through these food webs. Total Se concentrations in the biota were generally low (0.2 to 4.8 µg g(-1) dry weight) across sites and there was no relationship between distance from the coal-fired power plant and Se concentrations in invertebrates or fishes. Water chemistry was an equally poor predictor of Se concentrations in invertebrates and fish. Trophic position (determined using δ(15)N) was a significant predictor of Se concentrations in only five of the stream food webs, and two of these had negative slopes, indicating little or no trophic magnification across most systems; many fishes had lower concentrations than their invertebrate prey and trophic transfer was higher at sites with low invertebrate Se concentrations. Variability in Se concentrations in fishes was explained more by site of capture than microhabitat use within the site (as measured with δ(13)C), suggesting among-site differences in geological sources of Se. Because concentrations were below known toxicity thresholds for fish and other consumers, these results suggest that Se is not an environmental issue in New Brunswick streams that do not receive direct inputs from mining activities.

Antioxidant systems of brown trout (Salmo trutta f. fario) semen.

The present study characterizes the antioxidant systems of brown trout, Salmo trutta, semen as supplementation of semen dilution media with antioxidants can be beneficial to improve techniques for semen storage and cryopreservation. Antioxidants and oxidant defensive enzymes of spermatozoa and seminal plasma were analyzed. To determine whether antioxidants and oxidant defensive enzymes have an effect on sperm functionality, in vitro experiments were performed. Selected antioxidants and oxidant defensive enzymes were added to sperm motility-inhibiting saline solution and their effects on sperm viability (motility when activated, membrane integrity, and lipid peroxidation) were measured. In seminal plasma and spermatozoa the enzymes catalase, glutathione reductase, methionine sulfoxide reductase, peroxidase, and superoxide dismutase and the metabolites ascorbic acid, glutathione, methionine, tocopherol, and uric acid were detected. Of the enzymes superoxide dismutase had the highest activity, of the metabolites uric acid occurred in highest concentrations. During in vitro incubation uric acid and catalase increased the sperm motility, sperm membrane integrity, and decreased the sperm lipid peroxidation in comparison to the control. However, catalase was effective only at an activity much higher than that occurring in seminal plasma. Reduced methionine increased the sperm motility and sperm membrane integrity and oxidized methionine the motility. However, neither reduced nor oxidized methionine decreased the sperm membrane lipid peroxidation. It is concluded, that uric acid is the main antioxidant of brown trout semen.

The physiological stress response and oxidative stress biomarkers in rainbow trout and brook trout from selenium-impacted streams in a coal mining region.

Selenium (Se) is an essential element that can be toxic at concentrations slightly greater than those required for homeostasis. The main chronic toxic effects of Se in fish are teratogenic deformities, but Se can also activate the physiological stress response and redox cycle with reduced glutathione causing oxidative damage. Rainbow trout, Oncorhynchus mykiss, appear to be more sensitive to Se than brook trout, Salvelinus fontinalis. The objective of this study was to compare the physiological stress response (plasma cortisol, glucose, triiodothyronine, thyroxine, gill Na+/K+ ATPase, cortisol secretory capacity, K and liver somatic index) and oxidative stress biomarkers (liver GSH, GPx, lipid peroxidation, vitamin A and vitamin E) in rainbow trout (RNTR) and brook trout (BKTR) collected from reference and Se-exposed streams. The physiological stress response was not impaired (cortisol secretory capacity unchanged); although there were species differences in plasma cortisol and plasma glucose levels. Liver GSH, GPx and vitamin levels were higher in RNTR than BKTR, but lipid peroxidation levels were not different. The elevated GSH reserves may make RNTR more sensitive to Se-induced lipid peroxidation, but this may be offset by the RNTR's higher antioxidant (GPx and vitamin) levels. Species-specific biochemical differences may mediate differences in Se sensitivity and be used in aquatic Se risk assessments.

Effects of crude rapeseed oil on lipid composition in Arctic charr Salvelinus alpinus.

This study investigated the effects of crude rapeseed oil (RO) on lipid content and composition in muscle and liver of Arctic charr Salvelinus alpinus. Triplicate groups were fed diets containing fish oil (FO):RO ratio of 100:0, 75:25, 50:50 and 25:75 until two-fold mass increase. Total lipid content increased significantly in the liver with higher proportion of RO in the diet. Profound effects were seen in the fatty acid composition in the analysed tissues with a reduction in 20:5n-3 and 22:6n-3 and an increase in 18:2n-6 with higher RO content in the diets. A drop in cholesterol content was seen at 25% inclusion of RO in both tissues. Wild-caught fish contained a considerably higher amount of 20:4n-6 in both storage and membrane lipids of white muscle compared with the experimental fish.

The use of primary hepatocytes from brown trout (Salmo trutta lacustris) and the fish cell lines RTH-149 and ZF-L for in vitro screening of (anti)estrogenic activity of wood extractives.

Wood extractives are constituents of wood present in pulp and paper mill effluents, which may cause reproductive disturbances in fish. In the present study, we examined three cellular in vitro bioassays in order to assess (anti)estrogenic potencies of the wood extractives dehydroabietic acid (DHAA), isopimaric acid (IPA), betulinol (BET), hydroxymatairesinol (HMR), a phytosterol preparation (ULT), an oxidized phytosterol preparation (OX) and the model estrogen 17beta-estradiol (E2). The test systems used were primary hepatocyte cultures from brown trout and two piscine liver cell lines, RTH-149 and ZF-L. Estrogenicity was measured as vitellogenin (Vtg) secretion in cell culture medium. The primary hepatocytes cultures responded to E2 in a dose-dependent way. Vtg induction was inhibited with a simultaneous exposure to 4-hydroxytamoxifen (4-HT) indicating an estrogen receptor mediated response. DHAA and ULT induced a weak statistically non-significant Vtg production, and weak additive effects were found in some combination treatments of wood extractives and E2. Additionally, a pulp mill effluent tested on primary hepatocytes induced Vtg production when exposed at a 1% dilution. The cell lines secreted negligible amounts of Vtg upon E2 stimulation, which was neither dose-dependent nor inhibited by 4-HT. In conclusion, trout primary hepatocytes could be useful for assessing (anti)estrogenic potencies of compounds, and the wood extractives and a pulp mill effluent showed only weak or no estrogenic activity in this model system.