A comparative and evolutionary approach to oxidative stress in fish: A review

Oxidative stress results from an imbalance between the production of reactive oxygen species and the antioxidants defences, in favour of the former. In recent years, the association between oxidative processes, environmental change and life histories has received much attention. However, most studies have focused on avian and mammalian taxonomic groups, with less attention given to fish, despite their ecological and socio‐economic relevance. Here we present a review of the extrinsic and intrinsic factors that influence oxidative processes in fish, using a comparative and evolutionary approach. We demonstrate that oxidative stress plays a key role in shaping fish's responses to environmental change as well as life history strategies. We focus on representative examples to compare and contrast how levels of oxidative stress respond to changes in temperature, salinity and oxygen availability. Furthermore, we describe how emerging threats (i.e. pollution) affect oxidative stress parameters in fish. Oxidative stress indicators are increasingly being used as biomarkers to understand the mechanisms of various human‐induced stressors, but also to understand the physiological consequences of how animals are distributed in space and time and influenced by different life stages. Despite the expansion of the field of ecological oxidative stress, we are only beginning to understand the complex ways in which oxidative stress may interact with both extrinsic and intrinsic factors in fish. We conclude with a research agenda for oxidative research on fish and note that there is need for further research particularly in the area of life history strategies and ecological implications of oxidative status, as this type of research has the potential to help us understand patterns and dynamics relevant to fish conservation.     

Low levels of cadmium exposure induce DNA damage and oxidative stress in the liver of Oujiang colored common carp <Emphasis Type="Italic">Cyprinus carpio</Emphasis> var. <Emphasis Type="Italic">color</Emphasis>

Cadmium (Cd) compounds are widely distributed toxic environmental and industrial pollutants, and they may bring danger to growth and development of aquatic organisms. In China, the Oujiang color common carp, Cyprinus carpio var. color, is a very important fish, from an economic point of view, and is well used for fish culture in paddy fields. The purpose of this study was to show the low concentrations of cadmium-induced oxidative stress response and DNA damage in the livers of Cyprinus carpio var. color. Superoxide dismutase (SOD), malondialdehyde (MDA), and glutathione (GSH) in liver were measured after exposure to Cd levels (0.41, 0.52, 0.69, 1.03 and 2.06 mg/L, respectively) for 7 days and compared with the control groups. DNA damage, including indicators of damage percentage, DNA tail length (TL) and DNA tail moment (TM) were also analyzed by comet assays. Results showed that MDA and GSH levels in all treatment groups increased significantly relative to the controls (P < 0.01). Treatment with Cd at concentration of 0.41 mg/L increased SOD activity, while treatment with Cd at concentrations >0.41 mg/L inhibited SOD activities. DNA damage percentage, TL and TM also significantly increased when the Cd level was >0.41 mg/L. Positive correlations were also found between DNA damage levels and MDA levels (r = 0.74 for DNA damage percentage, r = 0.83 for TL, r = 0.84 for TM; P < 0.01 for all) as well as between GSH and MDA levels (r = 0.77, P < 0.01). These results strongly suggested that Cd-induced DNA damage in the livers of Cyprinus carpio var. color was due to lipid peroxidation and oxidative stress.     

氧化应激对鲤抗氧化状态和免疫功能的影响

为探讨氧化应激对鲤抗氧化状态和免疫功能的影响,本实验以H_2O_2作为活性氧自由基(ROS),将鲤暴露于不同浓度的H_2O_2(0、0.25、0.50和1.00 mmol/L)中,诱导鲤产生氧化应激反应。连续暴露7 d后,采集鲤血液和肝组织,以检测相关生化指标以及基因表达量的变化。结果显示,与空白对照组(0 mmol/L)相比,随着H_2O_2浓度的升高,血清葡萄糖(GLU)、皮质醇(cortisol)和乳酸(LA)含量显著升高;而碱性磷酸酶(AKP)和酸性磷酸酶(ACP)活性仅在1.00 mmol/L H_2O_2处理组中显著高于其他实验组。氧化应激参数显示,与空白对照组(0 mmol/L)相比,0.50和1.00 mmol/L H2O2处理显著降低血清过氧化氢酶(CAT)活性,而提高还原型谷胱甘肽(GSH)、丙二醛(MDA)和总抗氧化能力(T-AOC)水平;在肝脏组织中,1.00 mmol/L H_2O_2处理显著降低了GSH含量,促进了MDA生成。基因表达结果显示,与空白对照组相比,1.00 mmol/L H2O2处理组显著上调了肝脏组织中cyp1a表达,而下调了cyp1b表达;同时0.50和1.00 mmol/L H2O2处理显著上调了hsp70、hsp90、c3、c-lyz和hep的表达。研究表明,氧化应激暴露可诱导鲤产生明显应激反应和脂质过氧化,降低机体抗氧化能力并激发免疫应答反应。     

Cardiovascular responses to acute handling stress in the Antarctic fish Trematomus bernacchii are not mediated by circulatory catecholamines

Cannulated specimens of the benthic Antarctic teleost Trematomus bernacchii were subjected to a five min period of handling stress, following which blood pressures were measured and then blood samples taken for measurement of plasma catecholamine concentrations. There were two control groups of fish, one group of undisturbed fish at Scott Base, Antarctica and the other group of undisturbed fish measured in Christchurch.Plasma catecholamine concentrations were not changed following 5 min of acute handling stress. However, heart rate, ventral aortic blood pressure and haematocrit were elevated following handling, suggesting increased sumpathetic nerve activity. The low plasma catecholamine concentrations measured in all three groups of fish are similar to those found in other species of teleost at rest.The resting heart rates found in this series of experiments were approximately double those recorded in previous work on this species and this difference may be correlated with the length of time that the fish are held in captivity. The fish transported to Christchurch also had higher mean cell haemoglobin concentrations (MCHC) and lower plasma chloride concentrations.     

Effect of N-acetyl cysteine and glycine supplementation on growth performance,glutathione synthesis,and antioxidative ability of grass carp, <Emphasis Type="Italic">Ctenopharyngodon idella</Emphasis>

An 8-week feeding trial was conducted to evaluate the effect of N-acetyl cysteine (NAC) and glycine supplementation on growth performance, glutathione (GSH) synthesis, and antioxidative ability of grass carp, Ctenopharyngodon idella. Four practical diets were formulated: control, control + 0.2% NAC, control + 0.5% glycine, and control + 0.2% NAC + 0.5% glycine. Each diet was randomly assigned to quadruplicate groups of 30 fish (approximately 8.8 g). Weight gain and specific growth rate were significantly increased with the supplementation of NAC. Supplementation of NAC plus glycine significantly increased the feed efficiency. Glutathione peroxidase (GPx) and γ-glutamine cysteine synthase (γ-GCS) in plasma were significantly increased with the supplementation of NAC plus glycine. GSH in plasma increased and malondialdehyde (MDA) decreased in fish fed diets supplemented with NAC. Respiratory burst, superoxide dismutase (SOD), and catalase (CAT) activity were not affected by NAC or glycine. These results clearly indicated that NAC improved the growth performance and restored GSH of grass carp, supplemented NAC together with glycine enhanced GSH synthesis, and improved the antioxidative ability of grass carp.     

Role of exogenous glutathione in teleost fish and its effects on antioxidant defense responses in rainbow trout exposed to 3,3',4,4'-tetrachlorobiphenyl

Intraperitoneal injection of glutathione (GSH) is shown to effectively increase tissue glutathione content in two teleost species, the rainbow trout and the American eel. GSH uptake into teleost tissues is a distinct piscine feature of GSH biochemistry compared with mammalian species where little or not uptake occurs of exogenously added GSH. Enhancement and depletion of tissue GSH status was used as a tool to examine the effects of altered GSH content on other cellular antioxidants in 3,3,4,4-tetrachlorobiphenyl (TCB) exposed rainbow trout. -Tocopherol content was unchanged and thiobarbituric acid reactive substances (TBARS), an indicator of lipid peroxidation, changed little in liver and muscle of trout exposed to TCB and the various GSH treatments. However, -tocopherol levels in muscle and superoxide dismutase activities in the various tissues were significantly higher in controls and TCB-saline treated fish than in GSH deficient and lipoate supplemented TCB exposed trout. GSH status thus appears to affect some cellular antioxidant defenses in TCB exposed trout.     

Development of a modified cortisol extraction procedure for intermediately sized fish not amenable to whole-body or plasma extraction methods

The corticosteroid hormone cortisol is the central mediator of the teleost stress response. Therefore, the accurate quantification of cortisol in teleost fishes is a vital tool for addressing fundamental questions about an animal’s physiological response to environmental stressors. Conventional steroid extraction methods using plasma or whole-body homogenates, however, are inefficient within an intermediate size range of fish that are too small for phlebotomy and too large for whole-body steroid extractions. To assess the potential effects of hatchery-induced stress on survival of fingerling hatchery-reared Spotted Seatrout (Cynoscion nebulosus), we developed a novel extraction procedure for measuring cortisol in intermediately sized fish (50–100 mm in length) that are not amenable to standard cortisol extraction methods. By excising a standardized portion of the caudal peduncle, this tissue extraction procedure allows for a small portion of a larger fish to be sampled for cortisol, while minimizing the potential interference from lipids that may be extracted using whole-body homogenization procedures. Assay precision was comparable to published plasma and whole-body extraction procedures, and cortisol quantification over a wide range of sample dilutions displayed parallelism versus assay standards. Intra-assay  %CV was 8.54 %, and average recovery of spiked samples was 102 %. Also, tissue cortisol levels quantified using this method increase 30 min after handling stress and are significantly correlated with blood values. We conclude that this modified cortisol extraction procedure provides an excellent alternative to plasma and whole-body extraction procedures for intermediately sized fish, and will facilitate the efficient assessment of cortisol in a variety of situations ranging from basic laboratory research to industrial and field-based environmental health applications.     

The effects of dietary organic or inorganic selenium in rainbow trout (Oncorhynchus mykiss) under crowding conditions

In the present study, the effects of different sources of selenium (Se; sodium selenite or selenomethionine) supplementation on the growth and serum concentrations of oxidative stress markers [malondialdehyde (MDA), 8‐isoprostane, glutathione peroxidase (GSH‐Px) activity] and muscle Se, MDA and heat shock protein 70 (Hsp70) levels in rainbow trouts were evaluated. The fish (n = 360; 0 + years old) with initial average weight of 20 ± 0.8 g were randomly assigned to 12 treatment groups consisting of 3 replicates of 10 fish each in a 2 × 2 × 3 factorial arrangement of treatments (stocking densities, Se sources, Se levels). The fish were kept at low (25 kg m^?3) or high (100 kg m^?3) stocking densities and fed a basal (control) diet or the basal diet supplemented with either 0.15 or 0.30 mg of Se kg^?1 of diet from two different forms: sodium selenite or selenomethionine. High stocking density decreased weight gain, feed intake and feed conversion ratio (FCR) when basal diet was fed (P = 0.001). A linear increase in feed intake and weight gain and improvement in FCR were found in sodium selenite (P = 0.01)‐ or selenomethionine (P = 0.001)‐supplemented fish reared under crowding conditions. Serum and muscle Se levels and serum GSH‐Px activity increased (P = 0.001) linearly, whereas serum and muscle MDA concentrations and serum 8‐isoprostane decreased linearly as dietary sodium selenite (P = 0.01) or selenomethionine (P = 0.001) supplementation increased. Selenomethionine and sodium selenite supplementation decreased Hsp70 in the muscle of fish reared under crowding conditions (P < 0.05). Supplementation with Se improved growth and antioxidant status of fish and the effects of selenomethionine were relatively greater than sodium selenite in the crowded groups. Results suggest that crowding conditions cause significant detrimental effects in rainbow trout indicated by increased oxidative stress, reduced feed intake and body weight gain. ?t also indicates that dietary Se supplementation offers a feasible way of reducing the losses in performance of rainbow trout reared under crowding conditions. Selenomethionine seems to be more effective than sodium selenite and the higer dose in the present study also seems to be more effective than the lower dose.     

Effects of cypermethrin on antioxidant status,oxidative stress biomarkers,behavior, and mortality in the freshwater mussel <Emphasis Type="Italic">Unio elongatulus eucirrus</Emphasis>

In this study, the effects of cypermethrin on antioxidant status, oxidative stress biomarkers, behavior, and mortality in the freshwater mussel Unio elongatulus eucirrus were examined. Cypermethrin was applied at concentrations of 5, 10, 20, 40, 80, and 160 μg/L, for 1, 24, 48, 72, and 96 h. With increasing cypermethrin concentrations, the mussels showed significantly (P < 0.05) increased lipid peroxidation (MDA), which might be associated with decreased levels of reduced glutathione (GSH) and catalase (CAT) activity in the digestive glands and gills of the mussels. Negative correlations were observed between the MDA and the GSH and CAT levels after cypermethrin exposure, indicating a protective role of GSH and CAT against lipid peroxidation, and suggesting the use of these antioxidants as a potential biomarker of toxicity associated with contaminant exposure in freshwater mussels. In addition, at 5 μg/L, cypermethrin had no effect on the duration of the active and rest periods. All higher concentrations caused inhibition of the filtration activity by reducing the active periods and lengthening the rest periods. The active periods shortened as the cypermethrin concentrations increased, the reduction being 90% of the control at 160 μg/L. Rest periods were longest (237% longer than the control) at 160 μg/L. The 48, 72, and 96 h LC50 values of cypermethrin for mussels were determined as 96.50, 77.96, and 59.20 μg/L, respectively (P < 0.05). The results indicate that the insecticide cypermethrin has a harmful effect not only on nontarget aquatic arthropods and fish, but also on mollusks, although the sensitivity of mussels is less than that of fish. Being a general toxicant for aquatic life, cypermethrin should be used with great caution in agriculture to protect natural waters from contamination.     

低氧胁迫对鲻幼鱼生长、能量代谢和氧化应激的影响

为研究低氧胁迫对鲻幼鱼生长、能量代谢和氧化应激的影响,实验将其放在溶解氧(DO,mean±SE)含量分别控制在(1.66±0.41)、(4.35±0.53)、(7.03±0.36)mg/L的条件下养殖10 d,然后恢复至接近饱和溶解氧含量7.0 mg/L的条件下养殖30 d,研究其特定生长率、排氨率、耗氧率、氧氮比和血浆、肌肉、肝脏及鳃组织的总抗氧化能力(T-AOC)、过氧化物歧化酶(SOD)活力、抗超氧阴离子活力(ASOR)、丙二醛(MDA)含量、乳酸(LD)含量、总谷胱甘肽(T-GSH)、还原型谷胱甘肽(GSH)和氧化型谷胱甘肽(GSSG)含量。结果表明:低氧胁迫对鲻幼鱼的生长、能量代谢影响显著,较严重缺氧组鲻的体质量、特定生长率(SGR)、耗氧率和排氨率显著低于其他处理,不具有补偿生长的能力;而轻微缺氧组获得完全补偿生长。低氧胁迫对鲻氧化应激指标影响显著,胁迫结束时鲻通过提高某些抗氧化酶的活力来增强抗氧化能力,以提高其应对恢复正常溶解氧环境可能带来的氧化应激的能力,同时在恢复溶氧后鲻氧化应激反应也较强烈。在恢复溶解氧阶段,肝脏中GSH显著增加,说明鲻体内的保护机制被激活。肝脏和鳃中MDA的含量在低氧胁迫后与溶解氧含量呈明显的负相关性,在复氧30 d后仍然高于对照组,表明低氧胁迫加强鲻肝脏和鳃组织脂质过氧化反应。     

Revisiting redox-active antioxidant defenses in response to hypoxic challenge in both hypoxia-tolerant and hypoxia-sensitive fish species

It is not known whether changes in antioxidant levels always occur in fish in response to the oxidative stress that usually accompanies a hypoxic challenge. The studies of antioxidant responses to hypoxia in fish have mostly focused on very anoxia-tolerant species and indicate that there is an enhancement of antioxidant defenses. Here we present new data on redox-active antioxidants from three species, which range in their tolerance to hypoxia: the epaulette shark, threespine stickleback, and rainbow trout, together with a compilation of results from other studies that have measured oxidative stress parameters in hypoxia-exposed fish. The results suggest that in general, fish do not show an increase in redox-active antioxidant defense in response to oxidative stress associated with hypoxia. Rather, the changes in antioxidant defenses during hypoxia are very much species- and tissue-specific and are not linked to the level of hypoxia tolerance of the fish species.     

Effects of dietary selenium of organic form against lead toxicity on the antioxidant system in Cyprinus carpio

In this study was evaluated potential protective effect of organic selenium (Se) on heavy metal stress induced by lead (Pb) in Cyprinus carpio. For this reason, C. carpio was exposed to sublethal concentration of Pb (1.5 mg/L Pb(NO₃)₂) for 14 days. The fish were fed a basal (control; measured 0.55 mg/kg Se) diet or a basal diet supplemented with 2.50 mg/kg (measured 2.92 mg/kg Se) organic Se (Sel-Plex^®) during the experiment period. The variations in glutathione peroxidase (GSH-Px), glutathione S-transferase (GST) activities, and levels of reduced glutathione (GSH) with malondialdehyde (MDA) in liver and brain tissues of C. carpio were investigated in experimental groups. GSH levels in liver and brain tissues were significantly decreased by exposure to Pb. GST activity was significantly increased (p < 0.05) in liver tissue, but decreased in brain of treated fish by exposure to Pb. Also, GSH-Px activity was significantly increased in liver tissue, but decreased in brain of Pb-treated fish. Levels of MDA were increased in liver and brain of Pb-treated fish. The organic Se treatment for Pb-intoxicated animals improved activities of GSH-Px, GST and levels of MDA within normal limits. Supplemented Se could be able to improve Pb-induced oxidative stress by decreasing lipid peroxidation and regulating antioxidant defense system in tissues.     

Body growth, mitochondrial enzymatic capacities and aspects of the antioxidant system and redox balance under calorie restriction in young turbot (Scophthalmus maximus, L.)

Calorie restriction (CR) without undernutrition has been found to enhance stress resistance and life span in endotherms and ectotherms. We investigated the effect of 30% reduction in food offering on growth, aerobic capacities and oxidative stress parameters in young turbot (Scophthalmus maximus, L.). No differences in body weight, length and hepatosomatic index between the ad libitum (AL)-fed and the CR group occurred after 55 days of diet application. Of the mitochondrial marker enzymes, only citrate synthase (CS) activity in the liver was reduced under CR, whereas muscle CS activity and cytochrome oxidase (CCO) activity in both tissues remained the same in both feeding groups. The concentration of reduced glutathione increased significantly during feeding in muscle of CR fish, resulting in a more reduced glutathione redox ratio (GSH/GSSG) compared with AL fish muscle. Thiobarbituric acid-reactive lipid hydroperoxides (lipid peroxidation) but not protein carbonyl content (protein oxidation) was significantly reduced in CR fish muscle. Liver oxidative stress parameters did not vary significantly between experimental feeding groups. We conclude that 30% CR over 8 weeks has no adverse effect on young turbot. On the contrary, CR supports a reduced tissue oxidation state and reduces accumulation of lipid peroxidation products in muscle at sustained muscular aerobic capacity.     

The presence of uric acid, an antioxidantive substance, in fish seminal plasma

High concentrations of uric acid in seminal plasma of a range of teleost fish species are reported for the first time. Concentrations of urate amounted to 223.4; 121.9–130.0, 355.9, 735.6, 124.0, 192.7 and 148.0 M for rainbow trout (Oncorhynchus mykiss), yellow perch (Perca flavescens), muskellunge (Esox masquinongy), Northern pike (Esox lucius), carp koi (Cyprinus carpio), bream (Abramis brama), and tench (Tinca Tinca), respectively. Uric acid is an important anti-oxidant, therefore, it has a potential role in protection of fish spermatozoa against oxidative damage.     

Buprofezin toxication implicates health hazards in Nile tilapia (Oreochromis niloticus)

The objective of this study was to investigate the toxic effects of buprofezin insecticide on Nile tilapia (Oreochromis niloticus). The fish were exposed to buprofezin at 100 mg/L for 28 days. Compared to control, activity of serum transferases and levels of urea and creatinine showed significant increases. Oxidative stress was recorded manifested by elevated levels of malondialdehyde (MDA), reduced concentrations of reduced glutathione (GSH) and inhibition of activities of superoxide dismutase (SOD) and catalase (CAT) in liver and kidney. Examination of peripheral RBCs revealed elevated frequency of micronucleated cell. Interleukin 1 beta (IL‐1β) gene was upregulated in liver, muscle and brain, while that of cyclooxygenase 2 (COX‐2) gene increased in liver and muscle, but not in brain. Histopathological alterations were recorded in liver, kidneys, brain, gills, pancreas, spleen, intestine, muscle and ovaries. The immunohistochemical detection of caspase‐3 in the liver revealed no differences between treated and control groups; however, the expression of inducible nitric oxide synthase (iNOS) was demonstrated in hepatocytes and hepatopancreas in buprofezin‐treated group compared to control. It has been concluded that the tissue damage induced by buprofezin in Nile tilapia is mediated by oxidative stress and inflammatory response but not by apoptosis.     

Assessment of tissue-specific effect of cadmium on antioxidant defense system and lipid peroxidation in freshwater murrel, Channa punctatus

The effects of different concentrations of cadmium chloride on the extent of lipid peroxidation (LPO) and alterations in the antioxidant enzyme activities were studied in liver, kidney and gill tissues of freshwater murrel, Channa punctatus. The fish specimens were exposed to 6.7, 13.4 and 20.1 mg l⁻¹ sublethal concentrations of cadmium chloride and the oxidative stress was assessed after 24, 48, 72 and 96 h post-exposure. The biomarkers selected for the study were thiobarbituric acid reactive substances for assessing the extent of lipid peroxidation and antioxidant defense system such as reduced glutathione (GSH), glutathione reductase (GR), glutathione peroxidase (GP_X), glutathione-S-transferase (GST), catalase (CAT) and superoxide dismutase (SOD) activities. In general, the cadmium exposure elevated the LPO in subject tissues of treated group and modulated the activities of GPx, GST, SOD, CAT, GR and level of GSH after given exposure as compared to the control. All enzymes activities, except CAT (in kidney and gills), and amount of LPO elevated significantly (P < 0.05) in treated group with respect to control in all tissues, while significant difference was not observed between the exposed concentrations and within exposure duration. The results indicated that increase in LPO level and the fluctuation in antioxidant defense system in fish could be due to cadmium-induced increase in the production of reactive oxygen species (ROS). The potential role of these parameters as biomarkers of heavy metal pollution in aquatic system is discussed.     

Mammalian Growth Hormone-Releasing Hormone Indirectly Stimulates Sex Steroid Production in Tilapia

Abstract.— Fish possess a growth hormone similar to mammalian growth hormone, and the presence of a growth hormone-releasing hormone-like material in the brain has been demonstrated in several teleost species. We investigated the effect of a mammalian growth hormone-releasing hormone on serum estradiol-17β and testosterone concentrations in tilapia Oreochromis mossambicus and a hybrid O. niloticus × O aureus because growth hormone has been implicated in the regulation of gonadal functions in teleosts. Three hours after injecting the releasing hormone, male and female serum testosterone concentrations significantly increased compared to controls, while serum estradiol-17β concentrations increased significantly only in females. These increases were commensurate with respective serum steroid concentrations in fish injected with gonadotropin releasing-hormone. Studies utilizing hypophysectomized fish resulted in non-detectable levels of serum sex steroids in fish treated with the growth hormone-releasing hormone; however, significant increases occurred in fish treated with human chorionic gonadotropin. Similarly, gonadal tissue incubated with growth hormone releasing hormone had non-detectable levels of sex steroids in the media, whereas gonads incubated with human chorionic gonadotropin had significant increases in sex steroid concentrations. These studies suggest that growth hormone-releasing hormone acts at the level of the pituitary. Although mammalian growth hormones have been shown to increase serum gonadotropic hormone concentrations, this study provides evidence that a mammalian growth hormone-releasing hormone has the ability to indirectly increase sex steroid levels in fish.     

Stress responses and disease resistance in salmonid fish: Effects of chronic elevation of plasma cortisol

Basal levels of plasma cortisol in unstressed salmonid fish are normally in the range 0–5 ng ml⁻¹. An acute stress such as handling or 1 h confinement caused a temporary elevation of the plasma cortisol levels of both brown trout,Salmo trutta L., and rainbow trout,Salmo gairdneri Richardson, in the range 40–200 ng ml⁻¹ with a return to basal levels within 24–48 h. The extent of the cortisol elevation in response to an acute stress was dependent upon both the species and strain of trout. Chronic stresses, such as prolonged confinement or crowding, resulted in an elevation of plasma cortisol levels to approximately 10 ng ml⁻¹. Under these circumstances, blood cortisol levels remained elevated for periods of up to 4 weeks before acclimation finally occurred. It is shown, by means of intraperitoneal implantation of cortisol, that chronic elevation of plasma cortisol levels in the brown trout results in a dose-dependent increase in mortality due to common bacterial and fungal diseases. This effect is apparent at plasma cortisol levels as low as 10 ng ml⁻¹, levels below those often reported as being representative of ‘unstressed’ fish. These findings are discussed in relation to the known immunosuppressive effects of corticosteroids in teleost fish.