Trout Muscle Research Articles

Chloramphenicol Residues in Muscle of Rainbow Trout Following Two Different Dose Treatments

Depletion of chloramphenicol (CAP) in muscle of rainbow trout was evaluated following 4days of oral administration with two dosages (42 and 84mg/kg/day). Sampling was conducted during treatment and for 35days following the end of treatment. Analysis was carried out using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Concentrations observed during treatment were more than 300μg/kg. A significant elimination occurred within 9days after the cessation of treatment in both groups. Higher CAP levels were measured in the group treated with higher dose. CAP was not detectable after 13 and 15 after the end of treatment in both groups.

Effect of cold acclimation on troponin I isoform expression in striated muscle of rainbow trout

In vertebrates each of the three striated muscle types (fast skeletal, slow skeletal, and cardiac) contain distinct isoforms of a number of different contractile proteins including troponin I (TnI). The functional characteristics of these proteins have a significant influence on muscle function and contractility. The purpose of this study was to characterize which TnI gene and protein isoforms are expressed in the different muscle types of rainbow trout (Oncorhynchus mykiss) and to determine whether isoform expression changes in response to cold acclimation (4°C). Semiquantitative real-time PCR was used to characterize the expression of seven different TnI genes. The sequence of these genes, cloned from Atlantic salmon (Salmo salar) and rainbow trout, were obtained from the National Center for Biotechnology Information databases. One-dimensional gel electrophoresis and tandem mass spectrometry were used to identify the TnI protein isoforms expressed in each muscle type. Interestingly, the results indicate that each muscle type expresses the gene transcripts of up to seven TnI isoforms. There are significant differences, however, in the expression pattern of these genes between muscle types. In addition, cold acclimation was found to increase the expression of specific gene transcripts in each muscle type. The proteomics analysis demonstrates that fast skeletal and cardiac muscle contain three TnI isoforms, whereas slow skeletal muscle contains four. No other vertebrate muscle to date has been found to express as many TnI protein isoforms. Overall this study underscores the complex molecular composition of teleost striated muscle and suggests there is an adaptive value to the unique TnI profiles of each muscle type.

Proximate and Fatty Acid Composition in Muscle Tissues of Rainbow Trout (Oncorhynchus mykiss) Cultured in Yazd Province of Iran

Iran is the number one producer of cultured coldwater fish in Asia since 2005. Rainbow trout, (Oncorhynchus mykiss) is the most common and important fish produced by Iranian fisheries. There is not enough information about carcass composition of cultured fish in Iran. Rainbow trout muscle samples were collected from six fish farms of Yazd province during February 2008. Muscle samples were frozen at -30 ο C after being homogenized. Proximate composition of samples was measured. Saturated fatty acids including palmitic, stearic, myristic, lauric acids, also unsaturated fatty acids oleic and linoleic were extracted from muscle tissue of fish at different farms, using gas chromatograghy (GC). Other unsaturated fatty acids including α-linolenic acid, Eicosa Pentaenoic Acid (EPA) and Docosa Hexaenoic Acid (DHA) had low concentrations (up to 3 %) in samples. Vitamin E levels were 4.33 - 94.34 μg/100 g.

Adiponectin effects and gene expression in rainbow trout: an in vivo and in vitro approach

Here we present the presence of adiponectin and adiponectin receptors [type 1 (adipoR1) and type 2 (adipoR2)] in rainbow trout (Oncorhynchus mykiss) tissues and cell cultures together with the response to different scenarios. In response to fasting, adiponectin expression was up-regulated in adipose tissue, while the expression of its receptors increased in white and red muscle. Insulin injection decreased adipoR1 expression in white and red muscles. We deduce that the adipoRs in trout muscle show opposite responses to increasing insulin plasma levels, which may maintain sensitivity to insulin in this tissue. Adiponectin expression was inhibited by the inflammatory effect of lipopolysaccharide (LPS) in adipose tissue and red muscle. Moreover, results indicate that LPS may lead to mobilization of fat reserves, increasing adipoR1 expression in adipose tissue. The effects of LPS could be mediated through tumour necrosis factor α (TNFα), at least in red muscle. Insulin, growth hormone and TNFα all diminished expression of adipoR2 in adipocytes and adipoR1 in myotubes, while insulin increased the expression of adipoR2 in the muscle cells. Adiponectin activates Akt in rainbow trout myotubes, which may lead to an increase in fatty acid uptake and oxidation. Overall, our results show that the adiponectin system responds differently to various physiological challenges and that it is hormonally controlled in vivo and in vitro. To the best of our knowledge, this is the first time this has been demonstrated in teleosts, and it may be a valuable contribution to our understanding of adipokines in fish.

The effect of combining linseed oil and sesamin on the fatty acid composition in white muscle and on expression of lipid-related genes in white muscle and liver of rainbow trout (Oncorhynchus mykiss)

Sesamin (S) is a known lipid modulator and has been shown to increase the conversion of α-linolenic acid (ALA) to docosahexaenoic acid in rainbow trout (Oncorhynchus mykiss) fed vegetable oil mixtures including linseed oil. In this study, we evaluated the effects of S supplementation in linseed oil-based diets, content of α- and γ-tocopherols, fatty acid (FA) composition, as well as the gene expression of lipid-related genes. Fish with an average weight of 36.5 g were fed different combinations of commercial linseed oil (LO), purified linseed oil triacylglycerols (TAG) with polar fraction removed and a mixed linseed-sunflower oil (6:4 v/v) (MO). S was added at 0.58 g 100−1g feed and fed to the fish for a period of 58 days. Expression of PPARα was downregulated in white muscle of fish fed S containing diets (P < 0.05). The expression of PPARβ1A was not affected by S supplementation except where TAG oil was used. The expression of PPARβ1A declined significantly in TAG + S fed group (P < 0.05), which indicates that some minor compounds in linseed oil might suppress the effect of S on the expression of PPARβ1A. The expression of PPARγ(long) declined in LO + S and MO + S fed group (P < 0.05). The β-oxidation-related genes CPT1 and ACO were upregulated by vegetable oils compared to fish oil. S decreased percentage of ALA in white muscle of fish fed LO + S (P < 0.05). The increased desaturation index and the decreased ALA levels suggest that S may increase the biosynthesis of highly unsaturated FA in rainbow trout.

Time-dependent changes in protein expression in rainbow trout muscle following hypoxia

Adaptation to hypoxia is a complex process, and individual proteins will be up- or down-regulated in order to address the main challenges at any given time. To investigate the dynamics of the adaptation, rainbow trout (Oncorhynchus mykiss) was exposed to 30% of normal oxygen tension for 1, 2, 5 and 24 h respectively, after which muscle samples were taken. The successful investigation of numerous proteins in a single study was achieved by selectively separating the sarcoplasmic proteins using 2-DE. In total 46 protein spots were identified as changing in abundance in response to hypoxia using one-way ANOVA and multivariate data analysis. Proteins of interest were subsequently identified by MS/MS following tryptic digestion. The observed regulation following hypoxia in skeletal muscle was determined to be time specific, as only a limited number of proteins were regulated in response to more than one time point. The cellular response to hypoxia included regulation of proteins involved in maintaining iron homeostasis, energy levels and muscle structure. In conclusion, this proteome-based study presents a comprehensive investigation of the expression profiles of numerous proteins at four different time points. This increases our understanding of timed changes in protein expression in rainbow trout muscle following hypoxia.

Malachite green residues in farmed fish in Croatia

Malachite green concentrations were determined in muscle tissues samples of 42 carp (Cyprinus carpio) and 30 rainbow trout (Oncorhynchus mykiss) collected from fish farms in Croatia from 2009 to 2011. Concentrations were determined using the immunoassay method, validated according to the criteria of Commission Decision 2002/657/EC. The method performance is appropriate for the detection of malachite green, due to the obtained CCβ (0.68 μg kg−1) and LOD (0.31 μg kg−1) values that were below the MRPL level of 2 μg kg−1. Malachite green levels above the limit of detection of the method were determined in 18.1% of samples tested. The mean malachite green level (0.231 μg kg−1) of samples tested was lower than the determined limit of detection of the method. Concentrations ranged from 0.1 to 0.3 μg kg−1 in 68.1% of all fish samples. The measured concentrations exceeded 1 μg kg−1 in only two samples and the highest concentration (1.07 μg kg−1) was measured in rainbow trout muscle. Since this study is limited to malachite green residues, further investigation is required for determination of its metabolite, leucomalachite green. Additional surveillance programmes with a larger number of fish muscle samples should be carried out to improve the risk assessment.

Changes in carbonic anhydrase activity and gene expression of Hsp70 in rainbow trout (Oncorhynchus mykiss) muscle after exposure to some metals

The effects of some heavy metals Co(II), Cu(II), Zn(II), and Ag(I) on carbonic anhydrase (CA) activity from rainbow trout (RT) muscle were investigated. Moreover, the effects of these metals on the expression of heat shock protein 70 (Hsp70) gene in muscle tissue were examined by real-time quantitative PCR (RT-PCR) in muscle tissue after exposure to the metals at the end of 6, 12, 24, and 48 h. CA was purified with a specific activity of 2300 EU mg^{-1}, a yield of 19%, and 1080-fold. The molecular weights (Mw) of subunit and native enzyme were approximately 30 and 31 kDa, respectively. Optimum pH, stable pH, optimum temperature, activation energy (E\alpha), activation enthalpy (\DeltaH), and Q_{10} (the difference in activity of enzyme caused by the increment of 10 °C) value were determined. Apparent Michaelis constant (K_m), maximum reaction rate (V_{max}), and turnover rate of the enzyme (K_{cat}) values were 1.29 mM, 0.17 µmol min^{-1}, and 28.8 s^{-1}, respectively. The catalytic efficiency (K_{cat}/K_m) was 22.3. The heavy metals decreased in vitro CA activity. Inhibition mechanisms of the metal ions were noncompetitive, except for the Co(II) ion, which was competitive. The expression of the Hsp70 gene was increased in the presence of the metal ions. The expression level at the end of 48 h was the highest for all of the metals. Consequently, the in vitro inhibition rank order was determined as Co(II) > Zn(II) > Cu(II) > Ag(I). Interestingly, Ag(I) was the most effective metal ion on Hsp70 gene expression.

Assessment of mercury contamination in the Bílina River (Czech Republic) using indicator fish

The aim of the study was to determine mercury content in the muscle of indicator fish and to assess mercury pollution along the Bílina River, which is one of the most important tributaries of the Elbe River. A total of eight sites were chosen on the Bílina River for sampling. Indicator fish chub (Leuciscus cephalus L), roach (Rutilus rutilus L.) and brown trout (Salmo trutta m. fario L.) in the total numbers of 24, 26 and 27, respectively, were sampled at four locations, since at the remaining sites fish were absent. Mercury concentrations in the muscle of sampled indicator fish were measured using cold vapour atomic absorption spectrometry on an AMA 254 analyser. The highest mercury content (0.12 ± 0.027 mg·kg-1) was found in the muscle of roach at the Ústí nad Labem site and the lowest mercury content (0.04 ± 0.008 mg·kg-1) in the muscle of brown trout from the Březenec (the first upstream site) site. A significant difference (P &lt; 0.05) between the analyzed fish species was found only between chub (0.08 mg·kg-1) and brown trout (0.04 mg·kg-1) at the Březenec site. The priority of this study was to assess the mercury contamination of the Bílina River because this river flows through a heavy industrial activity in the region (especially production of petrochemicals, agrochemicals, sorbents, plasticizers and textile auxiliaries). Despite the fact that the Bílina is an extensively polluted river, the obtained mercury results were very low and did not exceed the limit of 0.5 mg·kg-1 set by Commission Regulation No. 1881/2006.

Comparative efficacy of candling and glass plate compression for detection of diphyllobothriosis in rainbow trout (Oncorhynchus mykiss) musculature

The efficiency of the direct candling technique on fillets (candling 1) was compared with examination of cuts 4 mm thick or less (candling 2) and glass plate compression for the detection of plerocercoids of Diphyllobothrium spp. in muscles of rainbow trout, Oncorhynchus mykiss. Application of the three procedures gave the following results (percentage of infected fish/percentage of isolated plerocercoids): candling 1: 40.9/22, candling 2: 29.5/18.8, glass plate compression: 29.5/59.2, and combination of candling 1 and 2: 70.5/40.8. The combination of the three techniques yielded 100% sensitivity: 44 infected fish were detected of 77 trout examined. When different regions of the musculature were compared using the three techniques, a high density of plerocercoids and the highest percentage of infection (90.9%; 40 infected trout) were detected in the ventral musculature. Candling 1, candling 2 and glass plate compression on the ventral musculature gave the following case numbers and percentages, respectively, for the total of 44 cases: 9 (20.5%), 9 (20.5%), and 22 (50%).

The role of growth hormone in growth, lipid homeostasis, energy utilization and partitioning in rainbow trout: Interactions with leptin, ghrelin and insulin-like growth factor I

The growth-promoting effects of in vivo growth hormone (GH) treatment were studied in relation to size and lipid content of energy stores including liver, mesentery, white muscle and belly flap in rainbow trout. In order to elucidate endocrine interactions and links to regulation of growth, adiposity and energy metabolism, plasma levels of GH, insulin-like growth factor I (IGF-I), leptin (Lep) and ghrelin, were assessed and correlated to growth and energy status. In addition tissue-specific expression of lepa1 mRNA was examined. Juvenile rainbow trout were implanted with sustained-release bovine GH implants and terminally sub-sampled at 1, 3 and 6 weeks. GH increased specific growth rate, reduced condition factor (CF) and increased feed conversion efficiency resulting in a redistribution of energy stores. Thus, GH decreased mesenteric (MSI) and liver somatic index (LSI). Lipid content of the belly flap increased following GH-treatment while liver and muscle lipid content decreased. Independent of GH substantial growth was accompanied by an increase in muscle lipids and a decrease in belly flap lipids. The data suggest that the belly flap may function as an energy buffering tissue during episodes of feeding and lean growth. Liver and muscle lipids were positively correlated to body weight, indicating a size-dependent change in adiposity. Hepatic lepa1 mRNA positively correlated to MSI and CF and its expression decreased following GH treatment, coinciding with decreased hepatic lipid content. Plasma Lep was positively correlated to MSI and belly flap lipid content, suggesting that Lep may communicate energy status. In summary, the observed GH tissue-specific effects on lipid metabolism in rainbow trout highlight the complex physiology of the energy reserves and their endocrine control.

Proteome modifications of fingerling rainbow trout ( Oncorhynchus mykiss) muscle as an effect of dietary nucleotides

A feeding study was conducted to determine the effect of dietary nucleotides (NT) on growth performance and muscle proteome profile of rainbow trout fingerlings. Five experimental diets according to different levels of supplemented nucleotides (0, 0.05, 0.1, 0.15 and 0.2%) were assayed in rainbow trout for 8 weeks. Each diet was randomly allocated to triplicate groups of fish with initial average weight of approximately 23 g. The percentage of body weight gain (WG) and feed efficiency (FE) of fish were better when the fish were fed 0.15–0.2% diets. At the end of feeding trial, fish fed the basal and 0.2% diets were subjected to proteomic analysis. Proteins of the muscle tissue were analyzed using two-dimensional electrophoresis and MALDI-TOF/TOF mass spectrometry. Dietary NT caused differential expression of muscle metabolic proteins including glyceraldehyde-3-phosphate dehydrogenase, creatine kinase, adenylate kinase, nucleoside diphosphate kinase, and triosephosphate isomerase. In addition to metabolic enzymes, troponin-T-1 as a structural protein was found to increase in abundance in the treated fish. The altered expression of both metabolic and structural proteins in fish fed NT may be related to higher growth rate in rainbow trout. These findings provide basic information to understand possible mechanisms of dietary NT contribution to better growth in rainbow trout.

Elimination of Chloramphenicol in Rainbow Trout Receiving Medicated Feed

Chloramphenicol muscle residue levels in rainbow trout were determined after oral administration of 84 μg kg-1d-1 of chloramphenicol for four days. Samples were taken one day before treatment and for 43 days after the treatment was over. Chloramphenicol was analysed using an in-house enzyme linked immunoassay (ELISA) validated against the criteria of the Commission Decision 2002/657/EC. Validation parameters confirmed that the method was appropriate for the detection of chloramphenicol at levels below the minimum required performance limit (MRPL) of 0.3 μg kg-1. The highest chloramphenicol levels were observed on the first day after the treatment had ended (144.3 μg kg-1). Elimination was significant over the first seven days; significant differences were detected between days 1 and 3 (p<0.001), 3 and 5 (p<0.001), and 5 and 7 (p<0.05). Chloramphenicol levels dropped below MRPL to 0.17 μg kg-1 on day 9 after the end of treatment. From day 11 to 43, chloramphenicol residues were detectable in a range from 0.091 μg kg-1 (highest) to 0.011 μg kg-1 (lowest). Our results indicate that trout muscle tissue could be compliant with health requirements for consumption 10 days after withdrawal from chloramphenicol treatment.

Elimination of malachite green residues from meat of rainbow trout and carp after water-born exposure

Uptake, accumulation and depuration of malachite green (MG) in muscle and skin of rainbow trout and carp were studied. The concentrations of MG and its mayor metabolite leuchomalachite green (LMG) were determined with liquid chromatography coupled with an absorbance and a fluorescence detector. Rainbow trout were exposed to 1 mg l − 1 of MG for 1 and 3 h and to a slightly increased concentration 1.5 mg l − 1 for 1 h. Changing the dose of MG in the bath did not significantly affect the residue levels in rainbow trout, but the duration of the exposure had a major influence. One day after exposure, the rainbow trout that were exposed three times longer contained 3 times more LMG and 4.5 times more MG than the trout in the one-hour bath. The level of MG in rainbow trout quickly fell below the decision limit (CCα) of 0.6 μg kg − 1 , while the level of LMG was still greater than 0.5 μg kg − 1 (CCα) at the end of the experiment (289 days after exposure). Carp were exposed to higher concentrations of MG, 2 mg l − 1 for 1 and 3 h , and 4 mg l − 1 for 1 h. Nevertheless, they accumulated less MG than rainbow trout and the elimination of LMG was slower. Because of the low water temperature (11 °C–15 °C), the carp metabolism, and therefore also LMG elimination, were slowed.

Increasing stocking density causes inhibition of metabolic–antioxidant enzymes and elevates mRNA levels of heat shock protein 70 in rainbow trout

The objective of this study was to determine the effects of stocking density on the activity of glucose 6-phosphate dehydrogenase (G6PD), 6-phosphogluconate dehydrogenase (6PGD) and glutathione reductase (GR) enzymes in liver, muscle, gill and kidney tissues of rainbow trout. Fish were reared at different stocking densities (15kg/m3, 20kg/m3, 25kg/m3 and 30kg/m3). After adaptation period of 30days, the experiment was carried out for two months. Stocking density of the control group was 15kg/m3. Increasing stocking density caused inhibition in the activities of the enzymes except for kidney G6PD and 6PGD. Activity of both pentose phosphate pathway enzymes unexpectedly increased only in kidney whereas inhibition was observed in other tissues. Since the most powerful and gradual attenuation was observed in muscle tissue for all enzymes, we performed quantitative Real Time PCR to examine the expression of heat shock protein (Hsp70) gene in muscle in order to understand whether the decrease in enzyme activities is associated with stress. The mRNA expression data showed that Hsp70 expression levels significantly elevated at 25kg/m3 and 30kg/m3 stocking densities. Overall results indicate that increasing stocking density blocks the activity of metabolic and antioxidant enzymes and causes considerable stress in rainbow trout. The most susceptible tissue to increasing stocking density was observed to be the muscle.

Regulation of LXR by fatty acids, insulin, growth hormone and tumor necrosis factor-α in rainbow trout myocytes

The liver X receptor (LXR) has recently been described in salmonids. In mammals, this receptor is already known as a transcriptional factor that regulates diverse aspects of cholesterol, fatty acid and carbohydrate metabolism in various tissues, including muscle. Here we examined LXR in trout myocytes. For this purpose, we analyzed LXR target gene expression and gene profile during myocyte development. In addition, we studied the transcriptional regulation of LXR by hormones, a pro-inflammatory mediator and unsaturated fatty acids. Trout myocytes were incubated with LXR agonists (T091317, 22(R)-hydroxycholesterol) and unsaturated fatty acids for 24h. Furthermore, differentiated myocytes were incubated with insulin and growth hormone (GH) for 3h, 6h and 18h, and with tumor necrosis factor-α (TNFα) for 24h. Samples were also obtained in various stages of cell differentiation. Our results demonstrate that lipoprotein lipase (LPL), fatty acid synthase (FAS), ATP-binding cassette transporter A1 (ABCA1), peroxisome proliferator-activated receptor-α and β (PPARα, β) are target genes of LXR in trout muscle. LXR agonists increased LXR expression, thereby indicating that this receptor is autoregulated. Unsaturated fatty acids downregulated LXR gene expression. This observation suggests a regulatory mechanism of these molecules on LXR-mediated fatty acid synthesis and uptake. TNFα did not modulate LXR gene transcription. Expression of the LXR gene was activated by insulin and GH. These results suggest that LXR may play a lipogenic role through insulin stimulation and a tendency to promote anabolic effects through GH on trout myocytes.

Effect of Frozen Storage Temperature on Quality-Related Changes in Rainbow Trout (Oncorhynchus mykiss)

The effect of frozen storage temperature on quality-related parameters of rainbow trout (Oncorhynchus mykiss) muscle was studied in the interval from −10 to −80°C on samples stored for 1 to 18 months. The following quantities were measured: drip loss, water holding capacity and water distribution, color, lipid oxidation (thiobarbituric acid-reactive substances, TBARS), and membrane stability (enzyme activity). No effect of temperature on drip loss, water holding capacity, water distribution, or membrane stability was observed for samples stored below −20°C, whereas storage at −40°C or lower compared to −30°C or higher resulted in a reduced level of secondary lipid oxidation (TBARS). No advantage was gained by using temperatures below −40°C for frozen storage of trout regarding any of the properties investigated.

Polybrominated diphenyl ethers and organochlorine compound levels in brown trout ( Salmo trutta) from Andean Patagonia, Argentina

Polybrominated diphenyl ethers (PBDEs), polychlorinated biphenyls (PCBs), DDTs and endosulfan residues were analyzed in muscle, liver, gonads, gills and stomach content of brown trout ( Salmo trutta) from the Andean Patagonia. PBDEs, PCBs and DDTs levels were positively correlated with lipid content, while less hydrophobic endosulfans showed a poor correlation. Endosulfan levels were about 99.9% of total contaminant (highest levels in liver 500 × 10 3 ng g −1 lipid weight), with α-/ β-isomers ratio >1 in all organs. These results are in agreement to the current-use of the technical endosulfan and also suggest acute exposure to this insecticide. Conversely, DDT/DDE ratio reflects fish exposure to old DDT sources, showing a DDE predominance. Gills had the highest levels of DDTs, PCBs and PBDEs, indicating they represent the main uptake pathway for such hydrophobic compounds from water column. PCBs showed the lowest levels in all organs and the PBDEs/PCBs ratios >1 agree with worldwide trends. PBDEs levels in gonads, gills, liver and muscle exceeded 80 ng g −1 (lipid weight) and were higher than other values reported in the Southern Hemisphere. BDE-47 was the predominant congener, suggesting higher bioaccumulation potential and possible brown trout metabolism of higher congeners. Since there is no point source of PBDEs in the region and residues were dominated by lower brominated congeners, atmospheric transport could be the main source of these compounds. This first report of PBDEs levels in fish from Argentina contributes to the knowledge about environmental trends of these persistent organic pollutants (POPs) in remote areas such as the Andean Patagonia.

Dietary carbohydrate-to-protein ratio affects TOR signaling and metabolism-related gene expression in the liver and muscle of rainbow trout after a single meal

Most teleost fish are known to require high levels of dietary proteins. Such high-protein intake could have significant effects, particularly on insulin-regulated gene expression. We therefore analyzed the effects of an increase in the ratio of dietary carbohydrates/proteins on the refeeding activation of the Akt-target of rapamycin (TOR) signaling pathways in rainbow trout and the effects on the expression of several genes related to hepatic and muscle metabolism and known to be regulated by insulin, amino acids, and/or glucose. Fish were fed once one of three experimental diets containing high (H), medium (M), or low (L) protein (P) or carbohydrate (C) levels after 48 h of feed deprivation. Activation of the Akt/TOR signaling pathway by refeeding was severely impaired by decreasing the proteins-to-carbohydrates ratio. Similarly, postprandial regulation of several genes related to glucose (Glut4, glucose-6-phosphatase isoform 1), lipid (fatty acid synthase, ATP-citrate lyase, sterol responsive element binding protein, carnitine palmitoyltransferase 1, and 3-hydroxyacyl-CoA dehydrogenase), and amino acid metabolism (serine dehydratase and branched-chain α-keto acid dehydrogenase E2 subunit) only occurred when fish were fed the high-protein diet. On the other hand, diet composition had a low impact on the expression of genes related to muscle protein degradation. Interestingly, glucokinase was the only gene of those monitored whose expression was significantly upregulated by increased carbohydrate intake. In conclusion, this study demonstrated that macro-nutrient composition of the diet strongly affected the insulin/amino acids signaling pathway and expression pattern of genes related to metabolism.

A Sensitive Analytical Method for Six Aflatoxins in Rainbow Trout Muscle and Liver

A highly sensitive analysis method for six aflatoxins (aflatoxin B₁, B₂, G₁, G₂, M₁ and aflatoxicol) in rainbow trout muscle and liver was developed. Aflatoxins (AFs) were extracted with acetonitrile-water (9 : 1), purified on an immunoaffinity column, and subjected to HPLC with fluorescence detection after post-column photochemical derivatization. The recoveries of AFs at 0.05 μg/kg spiking levels were 71.4-82.4% in muscle and 80.1-93.0% in liver, and the repeatability relative standard deviations (RSDr) were 0.87-4.6% in muscle and 2.0-6.2% in liver. Limits of quantitation (LOQs) and limits of detection(LODs)of AFs were estimated to be 0.004-0.029 μg/kg, and 0.002-0.012 μg/kg, respectively.