Rainbow Trout Liver Research Articles

Metabonomics analysis reveals the protective effect of nano‑selenium against heat stress of rainbow trout (Oncorhynchus mykiss)

Nano‑selenium (nano-Se) shows high biological activity and low toxicity, and has emerged as an ideal antioxidant. Our goal was to determine the underlying mechanism of nano-Se-mediated heat stress tolerance in rainbow trout (Oncorhynchus mykiss). Using liquid chromatography–mass spectrometry (LC–MS) metabolomics, histomorphology, and conventional biochemical assays, we investigated the physiological responses of heat-stressed rainbow trout to nano-Se. Fish were fed to two levels nano-Se at 18 °C for 9 days: CG18 (0 mg/kg) and Se18 (5 mg/kg). The water temperature of all groups was increased to 24 °C and maintained for 8 h (CG24, Se24). The results showed that most glycerophospholipids and CoA levels were decreased in CG18-CG24, and pathway enrichment analysis showed that it mainly interfered with glycerophospholipids and fatty acid metabolism. Meanwhile, hematoxylin and eosin and Oil Red O staining showed significant damage to CG18-CG24, which was ameliorated by Se18-Se24. The results combining analysis of antioxidant enzymes and heat shock proteins further support the notion that nano-Se supplementation inhibited galactose metabolism and activated the glutamate-glutamine metabolic pathway as the key metabolic strategy against heat stress. These results could establish heat stress defense strategies and increase our understanding of the mechanism of nutrient participation in fish's response to adverse environments. SignificanceGlobal warming is affecting the distribution and survival of cold-water fish worldwide, through seasonal water temperature increases and an increase in the frequency of extreme heat wave events. Surprisingly, Nano‑selenium (Nano-Se) with its outstanding advantages of high biological activity and low toxicity, making it a good Se nutrient supplement and free radical scavenger, and also an ideal and ecological way to supplement Se. How to utilize the metabolome to better address the complexity of the interactions that may occur with Nano-Se during the process of heat stress resistance is an important challenge. In the present study, this is the first publicly available metabonomics study of the anti-heat stress effect of Nano-Se as a nutrient on rainbow trout liver. These data indicated that Nano-Se effectively alleviated stress damage in rainbow trout, in which heat stress interfered with the metabolism of glycerophospholipids and fatty acids significantly, causing liver cell membrane damage and lipid metabolism disorders in rainbow trout. Meanwhile, supplementation of Nano-Se downregulated galactose metabolism and activated glutamate and glutamine metabolic pathways, which seems to be a key metabolic strategy to combat heat stress. The results provide a scientific basis for the development of an anti-heat-stress feed for rainbow trout that help maintain their health, productivity and welfare under unfavorable heat conditions.

New insights on in vitro biotransformation of anticoagulant rodenticides in fish

The assessment of the bioaccumulation potential of chemicals is an essential and mandatory part of their regulatory environmental risk and hazard assessment. So far, in vitro data on fish metabolism is rarely available for biocidal active substances such as anticoagulant rodenticides. In this case study we present in vitro biotransformation rates of eight biocidal and one pharmaceutical anticoagulants in rainbow trout (Oncorhynchus mykiss) liver subcellular S9 fraction (RT-S9) determined following the Organisation for Economic Co-operation and Development test guideline 319B method at two different incubation temperatures (i.e., 12 ± 1 °C and 23 ± 2 °C). Furthermore, we address challenges associated with the usability and interpretation of in vitro data to support the decision making within the regulatory bioaccumulation assessment in bridging the gap between in silico methods and in vivo studies. According to our results, four of the tested substances (i.e., chlorophacinone, coumatetralyl, bromadiolone, and difenacoum) exhibited significant intrinsic clearance (p < .001) in the RT-S9 assay. Overall, the observed metabolism was (very) slow and clearance rates were temperature-dependent. Whether the determined in vitro biotransformation rate had a substantial influence on the predicted bioconcentration factor during extrapolation was subject to the lipophilicity of the test substance. Further improvements of existing concepts are needed to overcome uncertainties in the prediction of bioconcentration factors for chemicals such as anticoagulants.

Assessment of pollution with biomarkers in fish culture.

Biotransformation enzymes are stimulated or inhibited depending on the exposure to xenobiotics and widely used as a biomarker. In this study, EROD activity, GST activity, reduced GSH amount, amount of lipid peroxidation product MDA as oxidative stress parameter, and ACHE inhibition as a neuromuscular parameter were measured in liver and muscle tissues of rainbow trout (Oncorhynchus mykiss) produced in aquaculture of Kemer Dam and in concrete ponds located in Fethiye. Samples were collected in January and July from fish farming in net cages in Kemer Dam and concrete cages in Fethiye in order to evaluate seasonal changes. Also, physicochemical properties of water samples collected from trout farms were measured and evaluated together with five biomarkers. Significant ACHE inhibition was observed in both stations in parallel with the increase in ammonium nitrogen in the July period (p ≤ 0.05). In parallel with the increase in turbidity in the Kemer Dam, a significant increase in GSH, MDA level, and GST activity was detected (p ≤ 0.05). As a result of the assessment of all data, significant increases were detected in EROD induction, GST activity, MDA level, and ACHE inhibition in concrete cages in Fethiye when compared to Kemer Dam (p ≤ 0,05). It was observed that pollution was at a lower level in July compared to January, and in Kemer Dam compared to Fethiye.

Evaluation of a full-scale wastewater treatment plant with ozonation and different post-treatments using a broad range of in vitro and in vivo bioassays

Micropollutants present in the effluent of wastewater treatment plants (WWTPs) after biological treatment are largely eliminated by effective advanced technologies such as ozonation. Discharge of contaminants into freshwater ecosystems can thus be minimized, while simultaneously protecting drinking water resources. However, ozonation can lead to reactive and potentially toxic transformation products. To remove these, the Swiss Federal Office for the Environment recommends additional "post-treatment" of ozonated WWTP effluent using sand filtration, but other treatments may be similarly effective. In this study, 48 h composite wastewater samples were collected before and after full-scale ozonation, and after post-treatments (full-scale sand filtration, pilot-scale fresh and pre-loaded granular activated carbon, and fixed and moving beds). Ecotoxicological tests were performed to quantify the changes in water quality following different treatment steps. These included standard in vitro bioassays for the detection of endocrine, genotoxic and mutagenic effects, as well as toxicity to green algae and bacteria, and flow-through in vivo bioassays using oligochaetes and early life stages of rainbow trout.Results show that ozonation reduced a number of ecotoxicological effects of biologically treated wastewater by 66 - 93%: It improved growth and photosynthesis of green algae, decreased toxicity to luminescent bacteria, reduced concentrations of hormonally active contaminants and significantly changed expression of biomarker genes in rainbow trout liver. Bioassay results showed that ozonation did not produce problematic levels of reaction products overall. Small increases in toxicity observed in a few samples were reduced or eliminated by post-treatments. However, only relatively fresh granular activated carbon (analyzed at 13,000 - 20,000 bed volumes) significantly reduced effects additionally (by up to 66%) compared to ozonation alone. Inhibition of algal photosynthesis, rainbow trout liver histopathology and biomarker gene expression proved to be sufficiently sensitive endpoints to detect the change in water quality achieved by post-treatment.

Investigating the Bioaccumulation Potential of Anionic Organic Compounds Using a Permanent Rainbow Trout Liver Cell Line

Investigating the Bioaccumulation Potential of Anionic Organic Compounds Using a Permanent Rainbow Trout Liver Cell Line

Metabolism and cytotoxicity of diphenylarsinic acid, a degradation product of sea-dumped chemical warfare agents, in a rainbow trout liver cell line RTL-W1.

Recent studies have found primary degradation products of phenylarsenic chemical warfare agents (CWAs) accumulating in fish tissues, while the potential effects of these dumped phenylarsenic CWAs, such as Clark I and II, in the Baltic Sea biota are poorly understood. In this study, the metabolism and cytotoxicity of diphenylarsinic acid (DPA), a primary degradation product of phenylarsenic CWA, was studied by incubating rainbow trout cell line RTL-W1 cells in media with 100mg/L DPA. Previously undescribed metabolites were identified by ultra-high performance liquid chromatography-high resolution mass spectrometry (UPHLCHRMS). Moreover, the cytotoxicity of diphenylarsine glutathione conjugate (DPA-SG), the major metabolite of DPA, was studied. Cytotoxicity of the compounds was evaluated using the Neutral Red retention test (NRR), showing an IC50 value of 278mg/L for DPA and 1.30mg/L for DPA-SG, indicating that the glutathione (GSH) conjugate of DPA is more than two orders of magnitude toxic than DPA itself, suggesting that toxic properties of DPA are increased after conjugation with intracellular GSH leading enhanced toxicity after uptake. Results gained in this study give more detailed information for elucidating biological effects of dumped chemical munitions in marine environment. Moreover, the results help in assessing the environmental and health risks posed by marine munition continued presence and deterioration in the sea bottom.

Effect of an Established Nutritional Level of Selenium on Energy Metabolism and Gene Expression in the Liver of Rainbow Trout.

The nutritional selenium (Se) has been demonstrated to have health-boosting effects on fish. However, its effect on fish energy metabolism remains unclear. This study explores the effect and underlying mechanism of the action of nutritional Se on energy metabolism in fish. Rainbow trout (Oncorhynchus mykiss) were fed a basal diet (0mg Se/kg diet) and a diet containing an already established nutritional Se level (2mg Se/kg diet, based on Se-yeast) for 30days. After the feeding experiment, the plasma and liver biochemical profiles and liver transcriptome were analyzed. The results showed that the present nutritional level of Se significantly increased liver triglyceride, total cholesterol, and plasma total cholesterol contents (P < 0.05) compared with the control. Transcriptome analysis showed that 336 and 219 genes were significantly upregulated and downregulated, respectively. Gene enrichment analysis showed that many differentially expressed genes (DEGs) were associated with lipid metabolism pathways (fatty acid biosynthesis, fatty acid elongation, and unsaturated fatty acid biosynthesis), carbohydrate metabolism pathways (glycolysis, the pentose phosphate pathway, and the citrate cycle), and the oxidative phosphorylation pathway. Real-time quantitative PCR (Q-PCR) validation results showed that the expression profiles of 15 genes exhibited similar trends both in RNA sequencing (RNA-seq) and Q-PCR analysis. These results reveal that optimum dietary Se activates glucose catabolic processes, fatty acid biosynthetic processes, and energy production and hence produces higher liver lipid content. This study concludes that the previously established level of nutritional Se (Se-yeast) (2mg/kg diet, fed basis) for rainbow trout promotes energy storage in the liver, which may benefit fish growth to some extent.

Levels of antioxidant enzyme activities in cultured rainbow trout (Oncorhynchus mykiss) fed with different diet compositions

The recent trend for rainbow trout diets to be higher in lipid content may increase the lipid concentration in fish and lipid peroxidation, which could lead to oxidative stress and affect fish health. In the present work, an evaluation of the possible effects of different fish diets on antioxidant enzyme levels was carried out on two aquaculture groups of rainbow trout (Oncorhynchus mykiss). The fish from one group were fed a diet containing 5% more crude lipids, than the diet used for the other group. The objective was to determine the effects of different concentrations of dietary lipid on the antioxidant defense enzymes superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), and glutathione-S-transferase (GST) in the liver and muscle of rainbow trout. All enzymes showed an increasing trend in fish fed with a diet containing more lipids. GST was observed to be the most sensitive antioxidant enzyme, followed by SOD and GPx, and finally by CAT and GR. These results provided data indicating the prooxidative effects of higher dietary lipid levels and suggested that dietary lipid plays an important role in determining fish susceptibility to oxidative stress.

Anoxia-reoxygenation modulates cadmium-induced liver mitochondrial reactive oxygen species emission during oxidation of glycerol 3-phosphate

Aquatic organisms are frequently exposed to multiple stressors including low dissolved oxygen (O2) and metals such as cadmium (Cd). Reduced O2 concentration and Cd exposure alter cellular function in part by impairing energy metabolism and dysregulating reactive oxygen species (ROS) homeostasis. However, little is known about the role of mitochondrial glycerol 3-phosphate dehydrogenase (mGPDH) in ROS homeostasis in fish and its response to environmental stress. In this study, mGPDH activity and the effects of anoxia-reoxygenation (A-RO) and Cd on ROS (as hydrogen peroxide, H2O2) emission in rainbow trout liver mitochondria during oxidation of glycerol 3-phosphate (G3P) were probed. Trout liver mitochondria exhibited low mGPDH activity that supported a low respiratory rate but substantial H2O2 emission rate. Cd evoked a low concentration stimulatory-high concentration inhibitory H2O2 emission pattern that was blunted by A-RO. At specific redox centers, Cd suppressed H2O2 emission from site IQ, but stimulated emission from sites IIIQo and GQ. In contrast, A-RO stimulated H2O2 emission from site IQ following 15 min exposure and augmented Cd-stimulated emission from site IIF after 30 min exposure but did not alter the rate of H2O2 emission from sites IIIQo and GQ. Additionally, Cd neither altered the activities of catalase, glutathione peroxidase, or thioredoxin reductase nor the concentrations of total glutathione, reduced glutathione, or oxidized glutathione. Overall, this study indicates that oxidation of G3P drives ROS production from mGPDH and complexes I, II and III, whereas Cd directly modulates redox sites but not antioxidant defense systems to alter mitochondrial H2O2 emission.

Effects of Vitamin C and E Supplementation on Growth, Fatty Acid Composition, Innate Immunity, and Antioxidant Capacity of Rainbow Trout (Oncorhynchus mykiss) Fed Oxidized Fish Oil

This 10-week feeding experiment examined the effects of supplementing vitamin C (VC) and E (VE) in oxidized oil diets on growth, fatty acid composition, blood physiological indicators, innate immunity, antioxidant capacity, and liver and intestine histology of rainbow trout (Oncorhynchus mykiss). Four diets were produced including a fresh fish oil containing diet (FFO diet), an oxidized fish oil containing diet (OFO diet), and OFO diet supplemented with 500 mg kg–1 VC and 400 mg kg–1 VE (OFO+C500+E400 diet) or 1,000 mg kg–1 VC and 800 mg kg–1 VE (OFO+C1000+E800 diet). Four hundred and twenty fish averaging 46.9 ± 0.32 g were stocked into 12 tanks and fed the diets twice a day to visual satiety. The results showed no significant effect of OFO or vitamins supplementation on growth, feed intake and feed utilization (P &amp;gt; 0.05). The groups that received OFO and OFO+C1000+E800 diets had significantly (P &amp;lt; 0.05) lower hepatosomatic indices than the other groups. Muscle fatty acid composition showed decreased proportion of 15:0, 16:0, 16:1n-7, 16:2n-4, 18:1n-5, 18:2n-6 (linoleic acid) and total saturated fatty acids in OFO group. Whereas higher percentages of 20:3n-6 and 20:3n-3 were found in OFO group. In addition, the highest ratio of docosahexaenoic acid (DHA, 22:6n-3) to eicosapentaenoic acid (EPA, 20:5n-3) was detected in OFO+C500+E400 group. The highest serum triglyceride concentration was recorded in the OFO group. Serum aspartate aminotransferase activity increased in OFO and OFO+C1000+E800 groups compared to OFO+C500+E400 group. Furthermore, significantly higher alkaline phosphatase activity in blood was found in OFO and OFO+C500+E400 groups. Significantly lower serum lysozyme, antiprotease, superoxide dismutase, and catalase activities were recorded in OFO group compared to FFO and OFO+C500+E400 groups, and an opposite trend was observed for malondialdehyde concentration. Muscle VC and VE concentrations, and liver and intestine histology remained unaffected. To conclude, feeding diet containing oxidized oil with peroxide value of 182 meq kg–1 with/without VC and VE supplementation did not influence growth and tissue VC and VE concentrations of rainbow trout. However, supplementing 500 mg kg–1 VC and 400 mg kg–1 VE reversed the adverse effects of OFO on non-specific immune response and antioxidant capacity.

Differential MHC-П, IL-8 and IgM genes expression in the liver, head Kidney and gill tissues of juvenile rainbow trout (Oncorhynchus mykiss) immunized by inactive trophonts of Ichthyophthirius multifiliis

Differential MHC-П, IL-8 and IgM genes expression in the liver, head Kidney and gill tissues of juvenile rainbow trout (Oncorhynchus mykiss) immunized by inactive trophonts of Ichthyophthirius multifiliis

Responses of microRNA and predicted mRNA and enzymatic targets in liver of two salmonids (Oncorhynchus mykiss and Salvelinus fontinalis) following air exposure.

The acute stress response is well-characterized, with rainbow trout as a teleost model for physiological and molecular responses. Air exposure, which stimulates an acute stress response, modulates liver microRNAs in rainbow trout; however, these highly conserved non-coding RNAs that bind to mRNA and repress translation, have never been measured in brook trout and it is unknown how miRNA expression responds following air exposure in this less studied salmonid. Our objective was to characterize the effects of air exposure on rainbow and brook trout liver miRNA expression, as well as the mRNA expression and enzyme activity that the miRNAs are predicted to target. Brook and rainbow trout were sampled pre- and 1-, 3-, and 24-h post- a three-minute air exposure. Plasma cortisol, glucose, and lactate were measured. Relative expression of miR-21a-5p, miR-143-3p, let-7a-5p and relative expression and enzyme activities of five predicted targets (pyruvate kinase, glucokinase, citrate synthase, cytochrome c oxidase, and catalase) were measured in liver. Rainbow and brook trout both had increases in plasma cortisol and lactate, while only rainbow trout had significant post-stress increases in plasma glucose. Furthermore, both trout species had increased miR-143-3p and miR-21a-5p relative expression 24-h post-stress. Four of the five enzymes measured had altered activity following stress. Brook trout miRNAs had inverse relative expression with relative catalase mRNA expression and cytochrome c oxidase enzyme activity, but no relationship was found in rainbow trout. Therefore, we have further characterized the transcriptional and enzymatic response to air exposure in two salmonids.

Biotransformation Potential of Cationic Surfactants in Fish Assessed with Rainbow Trout Liver S9 Fractions.

Biotransformation may substantially reduce the extent to which organic environmental contaminants accumulate in fish. Presently, however, relatively little is known regarding the biotransformation of ionized chemicals, including cationic surfactants, in aquatic organisms. To address this deficiency, a rainbow trout liver S9 substrate depletion assay (RT‐S9) was used to measure in vitro intrinsic clearance rates (CLint; ml min–1 g liver–1) for 22 cationic surfactants that differ with respect to alkyl chain length and degree of methylation on the charged nitrogen atom. None of the quaternary N,N,N‐trimethylalkylammonium compounds exhibited significant clearance. Rapid clearance was observed for N,N‐dimethylalkylamines, and slower rates of clearance were measured for N‐methylalkylamine analogs. Clearance rates for primary alkylamines were generally close to or below detectable levels. For the N‐methylalkylamines and N,N‐dimethylalkylamines, the highest CLint values were measured for C10–C12 homologs; substantially lower clearance rates were observed for homologs containing shorter or longer carbon chains. Based on its cofactor dependency, biotransformation of C12–N,N‐dimethylamine appears to involve one or more cytochrome P450–dependent reaction pathways, and sulfonation. On a molar basis, N‐demethylation metabolites accounted for up to 25% of the N,N‐dimethylalkylamines removed during the 2‐h assay, and up to 55% of the removed N‐methylalkylamines. These N‐demethylation products possess greater metabolic stability in the RT‐S9 assay than the parent structures from which they derive and may contribute to the overall risk of ionizable alkylamines. The results of these studies provide a set of consistently determined CLint values that may be extrapolated to whole trout to inform in silico bioaccumulation assessments. Environ Toxicol Chem 2021;40:3123–3136. © 2021 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Characterization of the ERK1/2 phosphorylation profile in human and fish liver cells upon exposure to chemicals of environmental concern

We developed phospho-ERK1/2 ELISA for human and rainbow trout liver cells, employing HepG2 and RTL-W1 cell lines as models. The assay was applied to detect changes in ERK1/2 activity for nine chemicals, added over a wide concentration range and time points. Cell viability was measured to separate ERK1/2 regulation from cytotoxicity. Perfluorooctane sulfonate and carbendazim did not change ERK1/2 activity; influence on ERK1/2 due to cytotoxicity was indicated for tributyltin and cypermethrin. Mancozeb, benzo[a]pyrene, and bisphenol A stimulated ERK1/2 up to ∼2- (HepG2) and 1.5 (RTL-W1)-fold, though the kinetics differed between chemicals and cell lines. Bisphenol A and benzo[a]pyrene were the most potent concentration-wise, altering ERK1/2 activity in pM (HepG2) to nM (RTL-W1) range. While atrazine and ibuprofen increased ERK1/2 activity by ∼2-fold in HepG2, they did not initiate an appreciable response in RTL-W1. This assay proved to be a sensitive, medium- to high-throughput tool for detecting unrecognized ERK1/2-disrupting chemicals.

Civa klorür (HgCl2) uygulanan gökkuşağı alabalığı (Oncorhynchus mykiss) karaciğer dokusunda nitrozatif ve oksidatif stresin rolü

The aim of this study was to determine oxidative stress caused by mercury chloride (HgCl2) in rainbow trout (Oncorhynchus mykiss) liver tissue. For this purpose, the LD50 value of HgCl2 on rainbow trout was determined as 551 μg/L. In the study, 40 fish in four groups were exposed to 25% and 50% (138 and 276 µg/L) of the two subletal doses of HgCl2 for 2 and 7 days, with 10 fish (n=10) in each group. To determine oxidative stress; peroxynitrite (ONOO−), total oxidant level (TOS), total antioxidant level (TAS), oxidative stress index (OSI) and malondialdehyde (MDA) were analyzed. In the study, it was observed that the differences between the groups in terms of ONOO−, TOS, TAS and OSI levels in the liver tissues was significant (P&lt;0.05), however, this difference was not significant (P&gt;0.05) in terms of MDA values. As a result, it can be concluded that HgCl2 increases ONOO−, TOS, TAS, OSI and MDA levels in liver tissue and even small doses of mercury are toxic to fish.

Cytochrome P450 Inhibition by Antimicrobials and Their Mixtures in Rainbow Trout Liver Microsomes In Vitro.

Antimicrobials are ubiquitous in the environment and can bioaccumulate in fish. In the present study, we determined the half-maximal inhibitory concentrations (IC50) of 7 environmentally abundant antimicrobials (ciprofloxacin, clarithromycin, clotrimazole, erythromycin, ketoconazole, miconazole, and sulfamethoxazole) on the cytochrome P450 (CYP) system in rainbow trout (Oncorhynchus mykiss) liver microsomes, using 7-ethoxyresorufin O-deethylation (EROD, CYP1A) and 7-benzyloxy-4-trifluoromethylcoumarin O-debenzylation (BFCOD, CYP3A) as model reactions. Apart from ciprofloxacin and sulfamethoxazole, all antimicrobials inhibited either EROD or BFCOD activities or both at concentrations <500 µM. Erythromycin was the only selective and time-dependent inhibitor of BFCOD. Compared with environmental concentrations, the IC50s of individual compounds were generally high (greater than milligrams per liter); but as mixtures, the antimicrobials resulted in strong, indicatively synergistic inhibitions of both EROD and BFCOD at submicromolar (~micrograms per liter) mixture concentrations. The cumulative inhibition of the BFCOD activity was detectable even at picomolar (~nanograms per liter) mixture concentrations and potentiated over time, likely because of the strong inhibition of CYP3A by ketoconazole (IC50 = 1.7 ± 0.3 µM) and clotrimazole (IC50 = 1.2 ± 0.2 µM). The results suggest that if taken up by fish, the mixtures of these antimicrobials may result in broad CYP inactivation and increase the bioaccumulation risk of any other xenobiotic normally cleared by the hepatic CYPs even at biologically relevant concentrations. Environ Toxicol Chem 2022;41:663-676. © 2021 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Comparison of EPA and DHA utilization in Atlantic salmon (Salmo salar) and rainbow trout (Oncorhynchus mykiss) fed two diets with different content of fish oil and rapeseed oil

ABSTRACT Atlantic salmon and rainbow trout with initial weights of 460–468 g were fed two diets either high in fish oil (FO) or rapeseed oil (RO) content. The abilities of the fish to metabolize and deposit lipid and n-3 fatty acids were compared. Growth rates, FCR, and fat digestibility were similar in both species. Rainbow trout showed higher condition factors and lower dress-out percentage and hepatosomatic index than salmon. Whole-body content of EPA and DHA were higher in both species fed the FO diet than in the RO diet, but indicated higher capacities to produce DHA in rainbow trout than in salmon. The total fat content in the RO group was lower and DHA higher in rainbow trout liver than in salmon, indicating a greater ability of rainbow trout to increase metabolism to DHA.

Adamantane carboxylic acids demonstrate mitochondrial toxicity consistent with oil sands-derived naphthenic acids

Naphthenic acids (NAs) are thought to be a primary cause of toxicity of oil sands process-affected water (OSPW). The purpose of this study was to determine if commercially available adamantane carboxylic acids act by mitrochondrial mechanisms similar to NAs found in OSPW. Mitochondria isolated from rainbow trout (Oncorhynchus mykiss) liver were exposed to commercially available adamantane acids, 3-hydroxyadamantane-1-carboxylic acid (CAS 42711-75-1) and 3,5-dimethyladamantane-1-carboxylic acid (CAS 14670-94-1), or to NAs extracted and purified from OSPW. The effects of these compounds on state 3 and 4 respiration, mitochondrial membrane potential, and mitochondrial reactive oxygen species (ROS; H2O2 production) were quantified. The compound 3-hydroxyadamantane-1-carboxylic acid only inhibited state 3 respiration at the highest concentration (2560 mg/L) and showed a concentration-dependent reduction in H2O2 production. Consistent with extracted OSPW NAs, 3,5-dimethyladamantane-1-carboxylic acid inhibited state 3 respiration and increased H2O2 production, but with a two-fold greater EC50 than the NA mixture extracted from OSPW. All three compounds uncoupled mitochondrial membrane potential and increased state 4 respiration. Based on these results, the adamantane 3,5-dimethyladamantane-1-carboxylic NAs may act via similar mitrochondrial mechanisms as NAs extracted from OSPW and could be used to further explore toxic mechanisms.

Effects of constant and diel cyclic temperatures on the liver and intestinal phospholipid fatty acid composition in rainbow trout Oncorhynchus mykiss during seawater acclimation

BackgroundRainbow trout is an economically important fish in aquaculture and is a model species in environmental physiology. Despite earlier research on the seawater adaptability of rainbow trout at different temperature regimes, the influence on the liver and intestine in this species is still unknown. Two trials were conducted to investigate the effects of constant and diel cyclic temperatures on phospholipid fatty acid (PLFA) composition in the liver and intestine of rainbow trout during seawater acclimation.ResultsAt the end of growth trial 1, fish at 9 and 12.5 °C showed significantly higher ratios of unsaturated to saturated (U/S) and unsaturation index (UI) than those at 16 °C in liver and intestine phospholipids. After day 1 of seawater acclimation, the U/S, UI, and average chain length (ACL) of liver and intestinal phospholipids in fish at 16 °C significantly increased. Two weeks after seawater acclimation, the liver and intestinal PLFA composition adapted to salinity changes. In trial 2, significantly higher U/S, UI, and ACL were found in intestinal phospholipids at 13 ± 2 °C. On the first day after seawater acclimation, UI and ACL in liver phospholipids significantly increased at 13 °C, while fish at 13 ± 2 °C showed significantly decreased U/S, UI, and ACL in the intestine. At the end of growth trial 2, liver PLFA compositions were stable, whereas intestinal PLFA at 13 and 13 ± 1 °C showed significantly decreased U/S, UI, and ACL. A two-way analysis of variance and principal component analysis revealed significant effects of different constant temperatures, seawater acclimation, and their interaction on the liver and intestinal phospholipids, a significant effect of diel cyclic temperature on intestinal phospholipids, and the effects of seawater acclimation and its interaction with diel cyclic temperature on liver phospholipids.ConclusionTemperatures of 9 and 12.5 °C could elevate membrane fluidity and thickness in the liver and intestine of rainbow trout in freshwater, whereas no significant effects were found with diel temperature variations. After seawater acclimation, constant and diel cyclic temperatures significantly influenced the membrane fluidity and thickness of the liver and intestine. Compared with constant temperature, diel temperature variation (13 ± 2 °C) can enhance the adaptability of rainbow trout during seawater acclimation.

Development of a microplate method for the determination of hepatic UDP-glucuronosyltransferase activity in rainbow trout (Oncorhynchus mykiss) and Nile tilapia (Oreochromis niloticus)

Hepatic glucuronidation represents an important phase II biotransformation reaction in both mammals and fish. The kinetic characteristics of uridine 5′-diphosphate (UDP) glucuronosyltransferases (UDPGTs) in rainbow trout liver microsomes were examined using p-nitrophenol (p-NP) as an aglycone and UDP-glucuronic acid (UDPGA) as a glucuronyl donor according to an existing protocol. The kinetic data obtained with varying concentrations of p-NP best fit the Hill equation and UDPGT activity was successfully induced following an i.p. injection of β-naphthoflavone (β-NF). The assay was subsequently adapted to a microplate method for determination of UDPGT activity in microsomal samples obtained from rainbow trout as well as Nile tilapia. In contrast to rainbow trout, UDPGT activity of Nile tilapia was best described by Michaelis-Menten kinetics. Based on the linearity of p-NP glucuronide formation, a p-NP concentration of 0.60 mM and a UDPGA concentration of 6.89 mM were determined to be suitable for assaying UDPGT activity in samples from rainbow trout and Nile tilapia. The microplate method offers several advantages over the historical assay; most notably it enables the observation of successive kinetics which ensures that enzyme activity is calculated in the most linear (initial) rate of the reaction. It also provides practical advantages in terms of ease-of-use and efficiency. This may be relevant to researchers investigating exposure of wild or farmed fish to environmental or feed-borne contaminants which are substrates of UDPGTs.