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

Fish welfare is an important issue for growth of the aquaculture industry. Stress responses represent animal's natural reactions to challenging conditions and could be used as a welfare indicator. Cortisol level is relevant to fish welfare condition, and is a readily measured component of the primary stress response system. Generally, cortisol is measured by blood sampling. However, fish blood cortisol level could be instantly influenced by handling-stress at sampling. Fecal corticoid metabolites (FCM) are a mixture of several different metabolites with a wide range of polarities. Thus, feces could be promising alternative less handling-sensitive and non-invasive biological matrices for cortisol evaluation in Atlantic salmon. In this study we developed non-invasive method for determination of fecal corticoid metabolites in farmed Atlantic salmon ( Salmo salar L.) using enzyme-linked immunosorbent assay (ELISA). It was demonstrated that salmon FCM extracted from salmon feces is insoluble in non-polar solvents like diethyl ether, but well soluble in polar solvents like methanol. The proper extraction ratio could be one ml 100% methanol for 100 μL of the liquid part of salmon feces or 100 mg of the solid part. The FCM directly detected in unextracted liquid part of feces correlated well with the FCM extracted from both liquid and solid part of the corresponding samples, without significant difference. Thus, it is feasible to measure FCM directly in the liquid part of salmon feces without any extraction procedure. Then, we applied this assay for FCM analysis in the group of salmon that experienced salmon pancreas disease (PD) and amoebic gill disease (AGD). We demonstrated 1) both plasma cortisol and FCM increased significantly during the outbreak of inflammatory disease (P < 0.01). Plasma cortisol level was elevated from 28 ± 40 ng/ml to 164.4 ± 62.5 ng/ml, FCM from 14.4 ± 13.2 ng/ml to 170.7 ± 89.7 ng/ml 2) Growth and starvation has no significant impact on either cortisol or FCM level. 3) FCM correlated well with plasma cortisol level (P < 0.01). Furthermore, there seems more individual variation in plasma cortisol levels than in FCM levels. These results suggest FCM could be directly analyzed in liquid part of salmon feces without extraction. This directly detected FCM level could represent the total fecal FCM level and plasma cortisol level. This simple and non-invasive method makes FCM a proper indicator for salmon welfare. [ABSTRACT FROM AUTHOR]

Lectins are a superfamily of carbohydrate-binding proteins that are widely distributed throughout living organisms. In earlier work, we identified lily-type lectin ( Sm LTL) in the skin mucus of turbot Scophthalmus maximus , and we characterized the protein in the present study. Results from qRT-PCR indicated that Sm LTL was expressed highly in skin, intestine and gill tissue. Changes in Sm LTL expression occurred in these tissues in response to environmental stressors including ciliate infection, high temperature and salinity. Recombinant Sm LTL purified from Escherichia coli was able to haemagglutinate mouse erythrocytes in the absence of calcium, and was inhibited by d -mannose. In addition, Sm LTL displayed selective binding to bacterial species including Edwardsiella tarda a nd Vibrio anguillarum , and exhibited toxicity towards Philasterides dicentrarchi, with a mortality of over 60% after 24 h at a concentration of only 100 μgml −1 . To investigate this toxicity further, we measured binding of Sm LTL after incubating the ciliate in FITC- Sm LTL solution. Surface fluorescence decreased substantially in the presence of 400 mM d -mannose. Together these results suggest that lily-type lectins serve as the first line of defence against microbial attack and play a pivotal role in the mucosal immune system. [ABSTRACT FROM AUTHOR]

Our study explored the dietary effects of adenosine monophosphate (AMP) to enhance growth, digestibility, innate immune responses and stress resistance of juvenile red sea bream. A semi-purified basal diet supplemented with 0% (Control), 0.1% (AMP-0.1), 0.2% (AMP-0.2), 0.4% (AMP-0.4) and 0.8% (AMP-0.8) purified AMP to formulate five experimental diets. Each diet was randomly allocated to triplicate groups of fish (mean initial weight 3.4 g) for 56 days. The results indicated that dietary AMP supplements tended to improve growth performances. One of the best ones was found in diet group AMP-0.2, followed by diet groups AMP-0.1, AMP-0.4 and AMP-0.8. The Apparent digestibility coefficients (dry matter, protein and lipid) also improved by AMP supplementation and the significantly highest dry matter digestibility was observed in diet group AMP-0.2. Fish fed diet groups AMP-0.2 and AMP-0.4 had significantly higher peroxidase and bactericidal activities than fish fed the control diet. Nitro-blue-tetrazolium (NBT) activity was found to be significantly (P < 0.05) greater in fish fed diet groups AMP-0.4 and AMP-0.8. Total serum protein, lysozyme activity and agglutination antibody titer were also increased (P > 0.05) by dietary supplementation. In contrast, catalase activity decreased with AMP supplementation. Moreover, the fish fed AMP supplemented diets had better improvement (P < 0.05) in body lipid contents, condition factor, hematocrit content and glutamyl oxaloacetic transaminase (GOT) level than the control group. Supplementation also improved both freshwater and oxidative stress resistances. Interestingly, the fish fed diet groups AMP-0.2 and AMP-0.4 showed the least oxidative stress condition. Finally it is concluded that, dietary AMP supplementation enhanced the growth, digestibility, immune response and stress resistance of red sea bream. The regression analysis revealed that a dietary AMP supplementation between 0.2 and 0.4% supported weight gain and lysozyme activity as a marker of immune functions for red sea bream, which is also inline with the most of the growth and health performance parameters of fish under present experimental conditions. [ABSTRACT FROM AUTHOR]

This study evaluated effects of graded levels of dietary β-glucan (0, 0.5, 1, 2 and 4 g kg −1 ) on growth performance, haematological parameters, intestinal Vibrio counts, dose requirement and salinity stress resistance in pompano fish Trachinotus ovatus (6.45 g ± 0.06 (SEM)). After 8-weeks of diet feeding, growth was significantly higher in fish fed diets with 0.10% β-glucan compared to fish fed control diet (no β-glucan). Survival increased significantly in fish fed 0.05 and 0.10% β-glucan compared to control diet. There were significant increases in red blood cells (in 0.20% β-glucan diet), in total leukocytes (in 0.05–0.20% β-glucan diet), in both lymphocyte and monocyte count in fish fed 0.10%–0.40% β-glucan diet. However, dietary β-glucan did not affect neutrophil, eosinophil and basophil counts. Intestinal Vibrio counts were reduced in fish fed any level of β-glucan compared to control. In addition, dietary β-glucan levels highly correlated with growth, survival, intestinal Vibrio counts and haematological index. Optimal β-glucan levels for maximal growth of fish were predicted to be 0.122% at day 21 (R 2 = 98.53%), 0.120% at day 28 (R 2 = 78.55%), 0.115% at day 42 (R 2 = 62.21%) and 0.090% at day 56 (R 2 = 75.18%), showing a decreasing β-glucan requirement with increasing fish size. Furthermore, optimal β-glucan levels for maximal haematological parameters based on lymphocyte count, was estimated to be 0.120% (R 2 = 98.53%) at day 56. Also, fish fed 0.05%–0.20% β-glucan showed better resistance against salinity stress. In conclusion, β-glucan supplementation is effective for improving growth, intestinal Vibrio counts and boosted stress resistance of the pompano fish, T. ovatus . [ABSTRACT FROM AUTHOR]

Yellow catfish ( Pelteobagrus fulvidraco ) has become a commercially important fish species in China and eastern Asia. High-density aquaculture has led to congestion and excessive stress and contributed to bacterial infection outbreaks that have caused high mortality. We investigated the effects of dietary supplementation with astaxanthin and emodin alone and in combination on the growth and stress resistance of yellow catfish. After 60 days of feeding, each group of fish (control, astaxanthin, emodin, and astaxanthin plus emodin (combination) groups) was exposed to acute crowding stress for 24 h, and a subsample of fish from the four groups was challenged with the bacterial septicemia pathogen Proteus mirabilis after the end of the crowding stress experiment. Compared with the control, the astaxanthin and emodin groups showed increases in serum total protein (TP), hepatic superoxide dismutase (SOD) activity and hepatic heat shock proteins 70 (HSP70) mRNA levels at 12 and 24 h after the initiation of crowding stress. The combination group exhibited increases in alanine aminotransferase (ALT) activity, aspartate aminotransferase (AST) activity, serum TP, hepatic SOD activity and hepatic HSP70 mRNA levels within 24 h after the initiation of crowding stress. However, decreases relative to the control were observed in the serum cortisol and glucose contents in the three treatment groups at 12 and 24 h after the initiation of crowding stress, in ALT and AST activity in the astaxanthin and emodin group at 24 h after the initiation of crowding stress, and in the serum lysozyme activity, serum alkaline phosphatase (ALP) activity, and hepatic catalase (CAT) and malondialdehyde (MDA) activity in the combination group at 24 h after the initiation of crowding stress. Additionally, the cumulative mortality after P. mirabilis infection was lower in all three treatment groups (57.00%–70.33%) than in the control (77.67%). Dietary supplementation with astaxanthin and emodin decreased the specific growth rate (SGR) and weight gain (WG) of healthy yellow catfish, although significant differences in mortality were not observed. These results indicate that dietary supplementation with 80 mg/kg astaxanthin and 150 mg/kg emodin can improve the anti-oxidative capabilities, hepatic HSP70 levels, and resistance to acute crowding stress of yellow catfish. Finally, an appropriate strategy for enhance yellow catfish stress resistance and disease resistance is proposed. [ABSTRACT FROM AUTHOR]

In this study, we show that induced spawning causes stress, an intense loss of epithelia and immunosuppression, decreasing physical and humoral protection in fish, effects that were prevented or improved in fish bathed with Aloe vera . A. vera has several medicinal properties, including wound healing and immunostimulatory effects, which we observed in this study. Fish bathed with A. vera had a higher number of epidermal goblet cells and, in general, an improved wound healing rate compared with the control after induced spawning. These effects might be related to (1) the stimulation of leukocyte activity, represented here by the increased leukocyte respiratory activity triggered by A. vera (leukocytes are recognized as playing an important role in wound repair); (2) the antimicrobial properties of A. vera, which decrease wound infection and accelerate the healing process; and (3) several mechanisms that explain the healing effect of A. vera (increased collagen synthesis, rate of epithelialization, and anti-inflammatory and moisturizing effects). Our results also suggest that caution is necessary during the induced spawning process, especially during stripping, and A. vera bathing is recommended after intensive aquaculture operations. [ABSTRACT FROM AUTHOR]

Fish is highly affected by many environmental stresses such as temperature and invasive infection. The extracellular signal-regulated kinase (ERK) pathway, part of the mitogen-activated protein kinase (MAPK) family, is found to act as crucial mediators for cell differentiation, proliferation and cell response to various stresses. In the present study, ERK2 ( Lc ERK2) and ERK5 ( Lc ERK2) were cloned and characterized from large yellow croaker, Larimichthys crocea . The full length cDNA sequence of Lc ERK2 was of 1910 bp, including an ORF of 1110 bp encoding a polypeptide of 369 amino acids. The full length cDNA sequence of Lc ERK5 was of 3720 bp, including an ORF of 3375 bp encoding a polypeptide of 1124 amino acids. Multiple alignments showed that both Lc ERK2 and Lc ERK5 contained highly conserved TEY motif and S_TKc domain in MAPK family and the unique catalytic and active structures of ERK2 and ERK5. Subcellular localization revealed that both Lc ERK2 and LcERK5 expressed in the cytoplasm and cell nucleus. The expression of Lc ERK2 and Lc ERK5 were detected in most tissues of large yellow croaker, with the most predominant expression of Lc ERK2 in brain and Lc ERK5 in gill, and the weakest expression of Lc ERK2 in liver and Lc ERK5 in intestine, respectively. The expression levels of Lc ERK2 and Lc ERK5 after temperature stress and poly I:C and flagellin challenge were investigated in LCK ( L. crocea kidney) cells. After temperature stress, significant down-regulations of Lc ERK2 transcripts were detected after 10 °C stress ( p < 0.05) whereas Lc ERK2 transcripts increased significantly after 35 °C stress ( p < 0.05). However, significant down-regulations of Lc ERK5 expression were detected at most time points after both cold and heat stress ( p < 0.05). However, significant up-regulations of Lc ERK2 and Lc ERK5 transcripts were found after immune challenge ( p < 0.05). Our results showed that Lc ERK2 transcripts enhanced after heat stress and both Lc ERK2 and Lc ERK5 transcripts could be induced by immune challenge. These findings indicated that Lc ERK2 might be more important in fish response to high temperature stress and both Lc ERK2 and Lc ERK5 might play an important role in fish immune response. [ABSTRACT FROM AUTHOR]

The aim of this study was to evaluate immunological, metabolic and osmoregulatory secondary stress responses in Eleginops maclovinus specimens submitted to three different stocking densities: i) low (3.1 kg m −3 ), medium (15 kg m −3 ) and high (60 kg m −3 ) during 10 days, alone or in combination with a previous treatment of a protein extract of the pathogen Piscirickettsia salmonis (0.5 μg g weight body −1 ). Plasma, liver, gill and kidney samples were obtained at the end of both experiments. Plasma cortisol and amino acid levels increased, while plasma glucose, triglyceride and lactate levels decreased at higher stocking densities. However, no effects were observed on serum Immunoglobulin type M (IgM anti P. salmonis level) values. Gill Na + , K + -ATPase activity enhanced under these experimental conditions, suggesting an osmotic imbalance. Energy metabolism changes, assessed by metabolite concentrations and enzyme activities, indicated a reallocation of energetic substrates at higher stocking densities. Specimens inoculated with a protein extract of P. salmonis and maintained at different stocking densities showed primary stress response, as all groups enhanced plasma cortisol concentrations. Serum IgM levels increased after treatment with P. salmonis extract but a negative influence of high stocking density on IgM production was observed when immune system was activated. Furthermore, treatment with P. salmonis protein extract evoked deep changes in the metabolite stores in all tissues tested, indicating a mobilization of energy substrates in response to infection. The results show that stocking density induced immunological, metabolic and osmoregulatory secondary stress responses in E . maclovinus specimens and that previous treatment with P. salmonis compromise these changes. [ABSTRACT FROM AUTHOR]

Calmodulin (CaM) is a multifunctional calcium sensor protein that participates in various cellular processes under normal, stress and pathological conditions. In crabs, however, the involvement of CaM in response to environmental stress and immune challenges has not been revealed yet. In the present study, a CaM cDNA ( EsCaM ) was identified from Chinese mitten crab Eriocheir sinensis and its mRNA expression patterns in response to ambient (salinity and pH) stress and immune challenges was examined. EsCaM encodes a 149-amino-acid protein with a calculated molecular mass of 16.8 kDa and an isoelectric point of 4.09. In unstimulated healthy E . sinensis , EsCaM mRNA transcript was detected in all tested tissues with predominant expression in hepatopancreas and the lowest expression in haemocytes. Ambient salinity (15‰ and 30‰ salinities) and pH (pH 6 and 8.5) stress significantly altered EsCaM mRNA expression in gill, hepatopancreas, haemocytes, intestine and muscle in Chinese mitten crab. In addition, EsCaM gene expression was significantly and rapidly induced as early as 2 h after LPS and Poly(I:C) immune stimulations in haemocytes in vitro . Furthermore, EsCaM expression was significantly up-regulated in E . sinensis haemocytes, gill, hepatopancreas, intestine and muscle in response to Edwardsiella tarda and Vibrio anguillarum challenges. Collectively, our findings suggest that EsCaM is an important stress and immune response gene in Chinese mitten crab. [ABSTRACT FROM AUTHOR]

Wild or farmed abalone are regularly exposed to stressors, such as air exposure and handling. Immune and transcriptional responses as well as susceptibility to vibriosis of sexually mature or immature European abalone acclimated at 16 or 19 °C were determined following handling or air exposure. Hemocyte density and H2O2 production increased while hemocyte viability and phagocytic index decreased following handling. Air exposure induces a decrease of hemocyte density and phagocytic index. Measurement of the expression of genes implicated in general metabolic, immunological and stress responses in gills, foot-muscle and hemocytes by real time q-PCR suggested that both stressors lead to a metabolic rate depression, characterized by a general inhibition of transcription. Finally, following handling a Vibrio harveyi challenge enhances almost 100% mortality of sexually immature animals at 19 °C while it has been previously demonstrated that only mature are susceptible to vibriosis. [ABSTRACT FROM AUTHOR]

Abstract: Clam Ruditapes philippinarum is one of the important marine aquaculture species in North China. However, pathogens can often cause diseases and lead to massive mortalities and economic losses of clam. In this work, we compared the metabolic responses induced by Vibrio anguillarum and Vibrio splendidus challenges towards hepatopancreas of clam using NMR-based metabolomics. Metabolic responses suggested that both V. anguillarum and V. splendidus induced disturbances in energy metabolism and osmotic regulation, oxidative and immune stresses with different mechanisms, as indicated by correspondingly differential metabolic biomarkers (e.g., amino acids, ATP, glucose, glycogen, taurine, betaine, choline and hypotaurine) and altered mRNA expression levels of related genes including ATP synthase, ATPase, glutathione peroxidase, heat shock protein 90, defensin and lysozyme. However, V. anguillarum caused more severe oxidative and immune stresses in clam hepatopancreas than V. splendidus. Our results indicated that metabolomics could be used to elucidate the biological effects of pathogens to the marine clam R. philippinarum. [Copyright &y& Elsevier]

Abstract: Four suppression subtractive hybridization (SSH) cDNA libraries were constructed to identify differentially expressed salinity stress responsive genes of black tiger shrimp, Penaeus monodon exposed to low (3 ppt) salinity conditions. Forward and reverse SSH cDNA libraries were developed from the gill and gut tissues of shrimp and clones having inserts larger than 300 bp were unidirectionally sequenced. Based on the sequence homology search, the identified genes were categorized for their putative functions related to a wide range of biological roles, such as nucleic acid regulation and replication, immune response, energy and metabolism, cell signaling, cellular process, cytoskeleton and membrane structure, stress and osmoregulation. Gene expression levels in response to low salinity conditions at 2 weeks post salinity stress of thirteen selected differentially expressed genes identified from SSH cDNA libraries (14-3-3 like protein, crustin, lysozyme, arginine kinase, Na+/K+-ATPase α-subunit, intracellular fatty acid binding protein, cathepsin B, anti-lipopolysaccharide factor, ferritin, ubiquitin conjugating enzyme E2, calreticulin, innexin 2 and heat shock protein 21) were analyzed by RT-PCR. The highest gene expression levels were observed for Na+/K+-ATPase α-subunit (34.28-folds) in gill tissues, intracellular fatty acid binding protein (13.30-folds) in gut tissues and innexin 2 (14.43-folds) in muscle tissues respectively. The differential and significant levels of gene expression indicate the functional role of these genes in shrimp salinity stress adaptive mechanisms. [Copyright &y& Elsevier]

Abstract: Manila clam Ruditapes philippinarum is an important marine aquaculture shellfish. This species has several pedigrees including White, Zebra, Liangdao Red and Marine Red distributing in the coastal areas in North China. In this work, we studied the metabolic differences induced by Vibrio harveyi in hepatopancreas from White and Zebra clams using NMR-based metabolomics. Metabolic responses (e.g., amino acids, glucose, glycogen, ATP and succinate) and altered mRNA expression levels of related genes (ATP synthase, heat shock protein 90, defensin and lysozyme) suggested that V. harveyi induced clear disruption in energy metabolism and immune stresses in both White and Zebra clam hepatopancreas. However, V. harveyi caused obvious osmotic stress in Zebra clam hepatopancreas, which was not observed in V. harveyi-challenged White clams samples. In addition, V. harveyi challenge induced more severe disruption in energy metabolism and immune stress in White clams than in Zebra clams. Overall, our results indicated that the biological differences between different pedigrees of R. philippinarum should be considered in immunity studies. [Copyright &y& Elsevier]

Abstract: Previous studies showed that heat-shock protein 60 (HSP60) was known to function as a molecular chaperone and is an important factor in the innate immune system in mammals. However, little was known about the physiological relevance of HSP60 in marine invertebrates. This study focuses on long-term monitoring of the differential expression of LvHSP60 in shrimp Litopenaeus vannamei in response to environmental stress. The thermal aggregation assay elucidated that LvHSP60 was an effective chaperone. It also suggested that LvHSP60 may employ the cell’s intrinsic mechanism to start the immunizing process. Using quantitative real-time PCR to monitor gene expression showed that LvHSP60 was variable under different stresses including environmental stress and pathogenic infection. LvHSP60 was speculated to regulate the adaptive responses to overcome environmental stresses. In conclusion, our study proved that LvHSP60 plays an important role in the intrinsic immune system and stress responses of shrimp. [Copyright &y& Elsevier]

Abstract: Hsp70 proteins are a family of molecular chaperones that are involved in many aspects of protein homeostasis. In this study, an Hsp70 homologue (SoHsp70) was identified from red drum Sciaenops ocellatus and analyzed at molecular level. The open reading frame of SoHsp70 is 1920 bp and intronless, with a 5′-untranslated region (UTR) of 399 bp and a 3′-UTR of 241 bp. The deduced amino acid sequence of SoHsp70 shares 84–92% overall identities with the Hsp70s of a number of fish species. In silico analysis identified in SoHsp70 three conserved Hsp70 domains involved in nucleotide and substrate binding. The coding sequence of SoHsp70 was subcloned into Escherichia coli, from which recombinant SoHsp70 was purified and, upon ATPase assay, found to exhibit apparent ATPase activity. Expressional analysis showed that constitutive expression of SoHsp70 was detectable in heart, liver, spleen, kidney, brain, blood, and gill. Experimental challenges with poly(I:C) and bacterial pathogens of Gram-positive and Gram-negative nature induced SoHsp70 expression in kidney to different levels. Stress-responsive analysis of SoHsp70 expression in primary cultures of red drum hepatocytes showed that acute heat shock treatment elicited a rapid induction of SoHsp70 expression which appeared after 10 min and 30 min of treatment. Exposure of hepatocytes separately to iron, copper, mercury, and hydrogen peroxide significantly upregulated SoHsp70 expression in time-dependent manners. Vaccination of red drum with a Streptococcus iniae bacterin was also found to induce SoHsp70 expression. Furthermore, recombinant SoHsp70 enhanced the immunoprotective effect of a subunit vaccine. Taken together, these results suggest that SoHsp70 is a stress-inducible protein that is likely to play a role in immunity and in coping with environmental and biological stresses. [ABSTRACT FROM AUTHOR]

Abstract: Differential leukocyte counts were taken of blood smears collected from laboratory stressed adult Australian swellsharks Cephaloscyllium laticeps. We calculated the granulocyte (combined count of heterophils and neutrophils) to lymphocyte (G/L) ratio to use as a new physiological indicator of stress for sharks. Animals were captured and stressed using commercial fishing gear (monofilament gillnet and mid-water longline) in a laboratory setting, with blood samples collected prior to capture and at predetermined intervals during a subsequent 72 h recovery period. There was a significant increase in the G/L ratio of 291.14 ± 54.13% at 72 h post-capture during recovery from the 6-h gillnet capture plus 15-mintue air exposure. Six hours of longline capture plus 15 min of air exposure also evoked a significant increase in the G/L ratio of 490.32 ± 294.25% (24 h post-capture) and 590.53 ± 277.65% (72 h post-capture). There was no significant change in the G/L ratio for control sharks that did not undergo capture stress but that experienced an identical blood-sampling regime as captured animals. Our study presents findings of stress-induced changes in leukocyte distribution within the peripheral blood of a shark species brought on by lymphopenia (decrease in lymphocytes) and granulocytosis (increase in granulocytes), and confirms this as a useful measure of the relatively rapid onset of stress in these animals. [Copyright &y& Elsevier]

Abstract: This study set out to investigate whether differential expression of genes for acute phase proteins in rainbow trout (Oncorhynchus mykiss) could be induced by confinement stress, a non-invasive method of activating the neuroendocrine stress response. In addition, a second objective was to assess the variation in gene expression between individual fish within the population of stressed fish in an attempt to identify APP genes having uniform and consistent changes in expression during stress. The liver was chosen for this investigation as it is the primary site of acute phase protein synthesis. Relative expression of the eight genes including transferrin, fibrinogen-like protein 2 (flp2), α-1-anti-proteinase-like protein (α1-antiprot), leukocyte cell-derived chemotaxin 2 (LECT2), pentraxin, serum amyloid A (SAA), haptoglobin (Hp), and differentially regulated trout protein 1 (DRTP1) was analysed by quantitative real-time PCR (qPCR) over 5 experimental time points spanning the course of a week. The results showed that the expression of three genes, SAA, haptoglobin and DRTP1, were most altered as a result of exposure to confinement stress. A correlation was identified between the expression of haptoglobin and DRTP1. Gene expression analyses in individual fish found that the transcript levels of haptoglobin and DRTP1 genes varied much less between individuals than was the case for SAA. The increase of haptoglobin and DRTP1 gene expression and its uniformity in response to stress make these genes potential biomarkers for stress in trout. [Copyright &y& Elsevier]