Effects of chronic confinement on physiological responses of juvenile gilthead sea bream, Sparus aurata L., to acute handling

Understanding how gilthead sea bream, Sparus aurata L., an important Mediterranean Sea species for aquaculture, respond physiologically to stressors commonly encountered in intensive rearing is important for effective production, as managing for stress is a major factor in maintaining healthy fish stocks. Our objective was to determine whether holding juvenile gilthead sea bream at a high density (HD), as a chronic stressor, would affect their physiological responses to a subsequent acute handling stressor. After acclimation at a low density (LD) of 6 kg m^?3 in 200‐L circular tanks containing 33–36 g L^?1 recirculating seawater at 19°C under a normal photoperiod, juvenile 37‐g gilthead sea bream were confined for 14 days at a HD of 26 kg m^?3 and then subjected to 30‐s aerial emersion in a dipnet. Plasma levels of cortisol, glucose, lactate, osmolality and chloride were determined in fish held in separate lots during LD (control) and HD confinement at 0, 1, 2, 7 and 14 days, and then after handling at 0, 1, 2, 4 and 8 h. Although plasma cortisol levels were similar in LD and HD fish groups after 14 d of confinement (15 and 23 ng mL^?1, respectively), the cortisol response in fish from the HD treatment at 1 and 2 h following acute handling (70 and 37 ng mL^?1, respectively) was only about half of that measured in the control group (139 and 102 ng mL^?1); plasma cortisol was similar in both groups by 4 and 8 h. In contrast, plasma glucose elevations in response to handling were higher at 4 and 8 h in the HD‐held fish (94 and 72 mg dL^?1, respectively) than in those from the LD treatment (59 and 51 mg dL^?1); glucose responses were similar in both groups at 1 and 2 h after handling and throughout confinement. Plasma lactate levels were higher in LD fish than in the HD group at the beginning of the experiment but were similar after 14 d confinement and responses to handling were similar (e.g. 33 and 35 mg dL^?1 at 1 h). Plasma osmolality showed increases during the first 2 h after acute handling but no differences were evident between the two density treatments at any time during confinement or posthandling. Plasma chloride levels did not change throughout the experiment. The reduced plasma cortisol response to acute handling likely resulted from negative feedback of mildly but chronically elevated circulating cortisol caused by the confinement stressor on the hypothalamic–pituitary–interrenal axis. While other post‐handling physiological changes also showed differences between treatment groups, the suppressed cortisol response in the HD‐held fish suggests a reduction in the gilthead sea bream's normal capacity to respond to an acute stressor.     

Effects of repeated crowding on the stress response and growth performance in Atlantic salmon (<Emphasis Type="Italic">Salmo salar</Emphasis>)

A 64-day growth experiment was conducted in which two groups of Atlantic salmon parr were grown under either control conditions or subjected to a weekly crowding stressor. Subjecting fish to the stressor resulted in a 7.7% reduction in wet weight after 29 days, which was maintained at 7.9% by day 64. This reduction in weight was reflected in a 44% reduction in specific growth rate and 38% increase in feed conversion ratio over the first 29 days of the experiment. Elevation in plasma cortisol was observed in crowded fish on days 1, 29 and 64. Similarly, on days 1 and 29 an increase in both plasma glucose and lactate was detected. On day 64, however, no differences in plasma glucose and lactate were observed, with the magnitude of the cortisol response also significantly reduced. Overall, the relatively moderate impact on growth performance and reduction in magnitude of measured stress parameters at the end of the experiment suggests possible habituation to the applied stressor.     

Modulation of the response of rainbow trout (Oncorhynchus mykiss Walbaum) to confinement, by an ectoparasitic (Argulus foliaceus L.) infestation and cortisol feeding

The effects of a 21 day infestation with the ectoparasite Argulus foliaceus on the response of rainbow trout Oncorhynchus mykiss to an acute stressor (confinement) were investigated. Fish were divided into a control and two parasitized groups, with one parasitized group having received cortisol feeding 72 and 24 h before infection. Prior to confinement only control fish showed a significant increase in body mass and condition factor. Following 4 h confinement, plasma glucose levels in the normal- fed/infested group were significantly higher than in the controls, while the cortisol-fed/infested group had significantly lower plasma levels of adrenocorticotropic hormone (ACTH) and glucose. Both infested groups, but not controls, had higher plasma ACTH and cortisol levels 48 h after confinement when compared with their pre-confinement levels. Stressor-induced immunomodulation was more evident in the parasitized fish after confinement, as was concluded from decreased lysozyme activity, alternative complement activity and oxygen radical production either immediately after, or 48 h post 4 h confinement. Circulating lymphocyte numbers were significantly reduced in the control and normal-fed/infested fish. Results indicate that low ectoparasitic infestations can, for a prolonged period, lead to a stress experience with effects that impair the response to subsequent stressors. Such effects may be further confounded by a previous elevation of plasma cortisol levels.     

Depletion of serum alternative complement pathway activity in gilthead seabream caused by α-tocopherol and n-3 HUFA dietary deficiencies

Studies on the alternative complement pathway (ACP) activity in fish have suggested the particular relevance of this pathway as a non-specific immune mechanism being its activation directly related with the availability of certain nutrients. In the present study ACP activity in gilthead seabream fed different dietary levels of a-tocopherol and n-3 HUFA deficiencies was studied under different stress conditions (overcrowding and repetitive chasing).A reduction in ACP activity was found when diets with deficiencies in -tocopherol and n-3 HUFA were assayed at any of the experimental conditions assayed. Levels of plasma cortisol were similar to those of fish after crowding stress subjected to dietary deficiencies at low stocking density, suggesting that dietary deficiencies produced a chronic elevation of cortisol. When fish were subjected to repetitive chasing, plasma cortisol from fish fed control diet showed a progressive increase whereas plasma cortisol levels from both dietary deficient groups showed a transitory peak 4 days after the beginning of the repetitive chasing. However, ACP activity levels of fish fed deficient diets remained lower than the control group and constant during chasing period. These results suggest that both a-tocopherol and n-3 HUFA nutritional deficiencies rather than stress are an important determinant of fish complement activity.     

Effect of stocking density on growth and serum concentrations of thyroid hormones and cortisol in Amur sturgeon, Acipenser schrenckii

This study investigated the effects of different stocking densities on growth and serum concentrations of thyroid hormones and cortisol in Amur sturgeon, Acipenser schrenckii. Fish were reared at low, medium, and high stocking densities (initial experimental densities were 0.30, 0.75, and 1.78 kg m⁻², respectively) for 70 days. The results showed that high stocking density had negative effects on growth and feeding efficiency, and altered serum levels of thyroid hormones and cortisol in Amur sturgeon. A significant decrease in specific growth rate was observed as stocking density was increased. The feeding rate decreased significantly in the medium and high density groups, indicating that high stocking density reduced the food consumption of sturgeon. Food conversion ratio increased with increasing stocking density, suggesting that high stocking density might inhibit fish growth through decreasing food conversion efficiency. Serum concentrations of total triiodothyronine, free thyroxine, and free triiodothyronine were inversely related to stocking densities, whereas serum total thyroxine level of sturgeon stocked at different densities remained stable. Also, higher stocking density resulted in an elevation of serum cortisol level, indicating that the sturgeon stocked at the higher density experienced density-dependent physiological stress. These results suggest growth suppression caused by high stocking density might be related to both crowding stress and the declines in peripheral circulating levels of thyroid hormones, as well as associated with the reductions in both food consumption and food conversion efficiency.     

Whole-body corticosteroid and plasma cortisol concentrations in larval and juvenile Atlantic cod Gadus morhua L. following acute stress

Methods were developed to assess whole‐body immunoreactive corticosteroid concentrations (IRC) and plasma levels of cortisol in Atlantic cod subjected to several common, acute stressors. A measurable corticosteroid stress response was observed at the first sampling in whole bodies of 8‐day post‐hatch (dph) larvae. Two groups of juveniles (～5 and 30 g) were subjected to a 30 s net stressor and whole‐body IRC and plasma cortisol levels were determined. Post‐stressor IRC in smaller fish rose approximately 14‐fold, peaked at 1 h, were sustained for 3–6 h and returned to pre‐stressor levels within 24 h. Post‐stressor plasma cortisol levels in larger fish rose approximately 18‐fold, peaked at 0.5–1 h, were sustained for 1–3 h and then returned to near pre‐stressor levels after 24 h. Immunoreactive corticosteroid concentrations appeared to remain elevated longer than plasma cortisol levels suggesting that steroids other than cortisol were contributing to total immunoreactivity in homogenates. Juveniles exposed to either a grading procedure or high density transport had maximal IRC and plasma cortisol levels within 90 min which returned to pre‐stressor levels within 24–72 h. A reduction in water salinity (20 g L^?1) did not moderate the corticosteroid response in juveniles. The results show that Atlantic cod respond to common, acute stressors in a manner similar to other teleosts. Whole‐body homogenates can be used to identify changes in IRC in response to acute stress in cod with the caveat that recovery IRC may differ from plasma cortisol concentrations.     

Stress responses of Atlantic salmon (Salmo salar L.) elicited by water level reduction in rearing tanks

The Atlantic salmon (Salmo salar) stress response was examined by measuring plasma cortisol, glucose and chloride in fish after water level reduction within rearing tanks. Maximum plasma cortisol levels (366±43 (SD) nM and 534±280 nM for Groups 1 and 2, respectively) were observed 20 min after application of the stressor. Cortisol levels were down to control levels 24 h later. The pattern for cortisol changes observed within these two groups were comparable. Plasma glucose and chloride concentrations did not change significantly in either of the experimental groups. The fish in group 2 were then repeatedly subjected to the same stressor every third day. After the fifth exposure to the stressor, blood was obtained from fish at times corresponding to sampling after the first exposure. Their maximum plasma cortisol level only reached 223 (±96) nM, and was down to prestress levels within 2 h. Plasma glucose and chloride concentrations did not change significantly in this second experiment. This challenge test revealed an acute primary response in Atlantic salmon without any apparent harmful secondary responses, that may thus serve as a standardized reference stressor using other fish groups under comparable conditions.     

Cortisol response of golden shiners (Notemigonus crysoleucas) fed diets differing in lipid content

The purpose of this study was to characterize the plasma cortisol response in golden shiners (Notemigonus crysoleucas) under crowding stress, and to determine whether dietary lipid affects the cortisol response. The main differences between the diets were the relative amounts and types of fatty acids of the n-3 and n-6 families. In experiment 1, triplicate groups of golden shiners were fed diets with 4 or 13% menhaden fish oil, or 4 or 13% poultry fat at 3% body weight for 4 weeks before the crowding stress. In experiment 2, fish were fed diets with 8% lipid from soybean oil (SBO), cod liver oil (CLO), (SBO+CLO), or cottonseed oil (CSO) for 6 weeks before the stress test. Plasma cortisol was measured initially, two hours after the stressor was applied, and two hours after the stressor was removed. The overall pattern of the cortisol response was similar to that seen in other warmwater fishes, but the rate and magnitude of the response were not affected by diet in either experiment. Golden shiners have an intermediate cortisol response compared to other fish that have been exposed to similarly stressful conditions.     

Effects of fasting on metabolic and immunological responses of common carp (Cyprinus carpio) to a further acute stress

Common carp were fasted for 0, 3 and 14 days prior to exposure to a 2‐hr crowding stress. Serum cortisol, glucose, lactate, _{T4, T3}, serum alternative complement (ACH50), lysozyme, total immunoglobulin (Ig) and blood WBC were determined before and 0, 6 and 24 hr after the crowding stress. Three days of fasting led to a significant decrease in serum levels of lactate and thyroid hormones. Fourteen days of fasting led to initiation of physiological process to maintain serum glucose by increase in serum cortisol level and lactate utilization. These fish had lower levels of thyroid hormones suggesting suppression of basal metabolism. The fish also had lower WBC and serum lysozyme and total Ig levels indicating immunosuppression. After the crowding stress, the fish fasted for 3 days had significantly lower lactate, thyroid hormones and total Ig levels. The fish fasted for 14 days had higher cortisol but lower glucose and thyroid hormones levels suggesting impairment in metabolic responses to the crowding stress. Immunological and health indicators including WBC, and ACH50, lysozyme and total Ig of these fish were significantly lower than the other groups after the crowding stress. Overall, the results show that a 14‐day fasting leads to stress responses, reduction in metabolism and immunosuppression. Moreover, 14 days of fasting lead to greater cortisol response to a further crowding stress, which induces marked immunosuppression and fallen welfare.     

拥挤胁迫对草鱼非特异性免疫功能的影响

在实验室条件下利用放养密度和放养时间构建组间差异,模拟养殖过程中草鱼所面临的拥挤胁迫。胁迫第7、14、30、60d后采样测量免疫指标。结果表明,高密度组(80g·L-1)的血清溶菌酶活性在胁迫后开始升高,并于14d时达峰值,以后逐渐下降,呈倒"U"型趋势。血清杀菌活性随放养密度的剧增而锐减(P<0.005)。白细胞比容在各组间未显示明显差异。而在30d和60d所测的高密度组草鱼的头肾巨噬细胞吞噬百分率和吞噬指数结果显示,高密度组低于对照组(P<0.05)。60d后用嗜水气单胞菌进行攻毒实验,高密度组死亡率高达75.0%,中密度组也达到了63.6%,而对照组只有25.0%。同期所测的脾脏脏器系数显示无论高密度组还是中密度组均显著低于对照组(P<0.01),且经解剖和正常鱼对比,发现其脾脏已明显萎缩。各实验组血清皮质醇浓度均在胁迫初期达到最大值,随后降低,至实验后期基本恢复到正常水平。     

Physiological stress responses to captivity in early developmental stages of the wedge sole Dicologoglossa cuneata (Moreau)

The stress responses in early growth stages of the wedge sole have been studied to determine whether the high cortisol levels described in juvenile fish are present from early developmental stages. Whole‐body cortisol, glucose and lactate contents, as well as biometric parameters in wedge sole larvae were measured at three different stocking densities. Stocking density affected growth‐related variables significantly, and larvae in lower stocking densities grew faster. Survival did not significantly differ among treatments. At hatching, the whole‐body cortisol concentration was 0.33 ± 0.01 ng g^?1 and varied significantly from 0 to 30 days after hatching (DAH) for each stocking density, though values remained stable for the remaining time in the low‐stocking density group. These hormone levels rose significantly (5.17 ± 2.43 to 22.10 ± 4.95 ng g^?1) at the end of the experiment, depending directly on the stocking density. Glucose and lactate‐body concentrations did not vary among treatments. We conclude that the stress responses of wedge sole larvae are detectable from 45 DAH and that stocking density already can be a stressor at that age. As described for juvenile stage, cortisol content values in wedge sole larvae under non‐stressful conditions are one of the highest among those reported in the literature. The captivity conditions could be responsible for this apparently stressful situation, though those values also could be normal in wild specimens.     

Reduction of stress response in carp,Cyprinus carpioL., held under deteriorating environmental conditions, by oral administration of bovine lactoferrin

Bovine lactoferrin (LF) was evaluated for its ability to suppress stress reactions in carp, Cyprinus carpio L., held under deteriorating environmental conditions. Three test diets containing different levels of LF (0.01, 0.1 and 1%) were fed at 2% of fish body weight per day, i.e. LF at 2, 20 and 200 mg kg(-1) body weight day(-1) was administered orally to carp for 14 days at 20 °C. In the fish reared at a density of 2 individuals 10 l(-1) , plasma cortisol, adrenaline and noradrenaline decreased in the groups fed with 0.1 and 1% LF content diets. In the fish held at a density of 10 individuals 10 l(-1) for 3 days, plasma cortisol, adrenaline, noradrenaline, dopamine and glucose were higher than those at 2 individuals 10 l(-1) in both the control and LF-treated groups. The extent to which these parameters were elevated in the fish held at the same density was reduced with increasing LF content of the diet. Twenty-four hours after exposure to hypoxic conditions (dissolved oxygen level, 2.5 mg l(-1) ), haemoglobin, haematocrit, plasma cortisol, adrenaline, noradrenaline and dopamine increased significantly in all groups. The responses of hypoxic fish fed with 0.1 and 1% LF diets for 14 days were smaller than those of the controls. These results suggest that LF has an ability to reduce the stress response of carp held under deteriorating environmental conditions.     

Reduction of transport stress of ayu by obligated schooling

We investigated stress responses of ayu Plecoglossus altivelis under the hypothesis that overcrowding during transport can be a stressor and the stress predisposes fish to disease by decline in immunocompetence. The extent of stress response was determined under different fish densities. As inferred from the serum concentration of cortisol 1 day after the treatment, the density at more than 23 kg/m³, which was within the range of normal transporting procedures, acutely evoked intense stress responses. To examine a possible way to reduce the stress response of fish under crowding conditions, we compared two fish groups established in water tanks with circulating (whirling) and turbulent currents at the same density. The cortisol levels 1 day after the treatment were 2.5-fold higher in turbulent current than in whirling current. Fish in whirling current formed a school, while those in turbulent current aggregated with each other. Schooling of fish induced by water flow may reduce their stress responses during transport. A transport system with a directional water flow should keep fish in good condition without reducing the number of fish contained.     

Effects of a probiotic bacterial <Emphasis Type="Italic">Lactobacillus rhamnosus</Emphasis> dietary supplement on the crowding stress response of juvenile Nile tilapia <Emphasis Type="Italic">Oreochromis niloticus</Emphasis>

To test the effects of probiotic bacteria against crowding stress, juvenile Nile tilapia Oreochromis niloticus were fed commercial feed supplemented with 1 × 10¹⁰ cfu/g pellets of Lactobacillus rhamnosus for 3 weeks. The fish were confined and subjected to crowding for 7 and 14 days after which their physiological condition was investigated. The administration of probiotic prophylaxis improved growth performance even in the stressed fish. After 7 days of stress exposure, fish receiving probiotic-supplemented feed showed proactive behavior and coping responses to the stressor, as evidenced by elevated plasma glucose levels and osmolality, stabilized plasma electrolytes, and a higher RNA:DNA ratio. Fish stressed for 7 days and fed normal commercial diet showed impairment of plasma electrolytes; after 14 days, the plasma osmolality, electrolytes, glucose, and RNA:DNA ratio decreased, indicating physiological maladaptation to the stressor. The growth rate was also found to be reduced, suggesting that energy demand exceeded the energy available from metabolism and dietary uptake. This latter effect was not observed in fish fed the probiotic-supplemented diet. The results suggest that probiotic bacteria prophylaxis may increase energy availability for metabolic support of the crowding stress response and improve the stress coping capacity of 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.     

Cortisol,glucose and hematocrit changes during acute stress,cohort sampling,and the diel cycle in diploid and triploid brook trout (Salvelinus fontinalis Mitchill)

In this study, groups of diploid (control) and triploid brook trout were subjected to a 5 min acute handling/confinement stress. Blood samples were collected from equal numbers of diploid and triploid fish at regular predetermined intervals, beginning prior to confinement and continuing for up to 3h after confinement. These blood samples were used for the measurement of hematocrit and plasma cortisol and glucose concentrations. The effects of cohort sampling and the diel cycle on these variables were also analysed. Except for minor differences, both groups responded similarly to the acute stress, with short-term elevations in plasma cortisol and hematocrit levels, and a more sustained increase in plasma glucose levels. No significant differences were found between diploid and triploid fish in their reactions to cohort sampling. However, the act of removing an individual from a tank caused significant changes in plasma cortisol levels in cohorts remaining in that tank (p < 0.001). Significant diel effects were observed for both ploidy groups in plasma cortisol (p < 0.05) but not in plasma glucose or hematocrit. To whom correspondence should be addressed     

Interactive effects of a high-quality protein diet and high stocking density on the stress response and some innate immune parameters of Senegalese sole Solea senegalensis

Amino acids (AA) regulate key metabolic pathways, including some immune responses. Therefore, this study aimed to assess whether an increased availability of dietary AA can mitigate the expected increase in plasma cortisol and metabolites levels due to high stocking density and its subsequent immunosuppression. Senegalese sole (Solea senegalensis) were maintained at low stocking density (LSD; 3.5 kg m^?2) or high stocking density (HSD; 12 kg m^?2) for 18 days. Additionally, both treatments were fed a control or a high protein (HP) diet (LSD, LSD HP, HSD and HSD HP). The HP diet slightly increased the levels of digestible indispensable AA, together with tyrosine and cysteine. HSD was effective in inducing a chronic stress response after 18 days of treatment since fish held at HSD presented higher plasma cortisol, glucose and lactate levels. Moreover, this increase in stress indicators translated in a decrease in plasma lysozyme, alternative complement pathway (ACP) and peroxidase activities, suggesting some degree of immunosuppression. Interestingly, while plasma glucose and lactate levels in HSD HP specimens decreased to similar values than LSD fish, plasma lysozyme, ACP and peroxidase activities increased, with even higher values than LSD groups for ACP activity. It is suggested that the HP diet may be used as functional feed since it may represent a metabolic advantage during stressful events and may counteract immunosuppression in sole.     

Plasma growth hormone levels in fed and fasted rainbow trout (Oncorhynchus mykiss) are decreased following handling stress

Plasma growth hormone concentrations of rainbow trout,Oncorhynchus mykiss, fasted for six weeks, were significantly (p < 0.01) higher than in comparable fed animals; in the fasted fish, the levels fell progressively following acute stress (by displacing the fish within their home aquarium), with significant differences from pre-stressed fish evident between one and thirty-two hours after application of the stressor. Plasma growth hormone concentrations also fell significantly in the fed group, but differences were evident only between two and eight hours after stressor application.Plasma cortisol concentrations in pre- and post-stressed fed and fasted fish were similar. There was a bimodal response to stressor application in both groups, with significantly higher values relative to the pre-stressed sample evident one and eight hours after disturbance, but not after two, four or thirty-two hours. The changes in plasma cortisol levels between the initial (09:00h) sample and the sample taken eight hours later resembles the diet pattern seen in trout given access to self-demand feeders.Plasma glucose concentrations in pre-stressed fed animals were higher than in pre-stressed fasted fish. This relationship was also evident between one and four hours and thirty-two hours after stressor application. The post-stress rise in plasma glucose concentration was evident between one and four hours in the fed group, and between four and eight hours in fasted fish.The diel changes in plasma growth hormone and glucose concentrations could not be attributed to normal circadian patterns, and there was no apparent correlation between changes in plasma growth hormone and cortisol concentrations. There was a significant inverse correlation between plasma glucose and growth hormone concentrations when the total data set were analyzed, but these correlations were not apparent when the treatment groups were analyzed separately.     

The Effect of Constant and Fluctuating Temperatures on the Confinement-induced Plasma Cortisol Stress Response in Channel Catfish lctalurus punctatus

Abstract.— Initial and confinement-induced changes of plasma cortisol were measured in channel catfish Ictalurus punctatus acclimated to upper and lower constant temperatures for each season, a spring/fall diurnal pattern between 15 and 26 C, and a summer diurnal pattern between 24 and 35 C. In diurnal treatments, temperature changes were achieved in 10 h and held constant for 2 h before reversing the temperature. Fish held in the spring/fall pattern or at a constant temperature of 15 or 26 C revealed no plasma cortisol concentration differences due to acclimation temperature, but hormone concentrations in all groups increased by about 4.5 fold following a 2-h confinement. Fish held on a summer pattern had differences in both the initial cortisol samples and those measured after confinement. Acclimation to a constant 35 C resulted in significantly higher initial cortisol concentrations, but these levels did not increase following confinement. The inability of the fish held at 35 C to respond to the confinement was thought to be due to the constant thermal stress, resulting in an inadequate interrenal reserve. Physiological consequences due to the diurnal physiochemi-cal cycling in ponds needs further attention.     

Physiological responses of juvenile wedge sole Dicologoglossa cuneata (Moreau) to high stocking density

Physiological responses to a high stocking density were tested in juvenile wedge sole Dicologoglossa cuneata (Moreau). Fish were kept at low (1 kg m⁻²), medium (3 kg m⁻²) and high (9 kg m⁻²) stocking densities for 22 days. No differences in the weight, length, survival and hepatosomatic index were observed among treatments. Basal plasma cortisol and osmolality were found to be directly and positively related to stocking density. A mild increase in plasma glucose was seen at medium density, and plasma protein was elevated at medium and high densities. The liver glucose and glycogen content was inversely related to stocking density. The liver triglyceride level was significantly elevated at the highest density, and the α-amino acid content decreased at the highest density. In muscle, glucose levels were significantly higher in fish kept at the lowest density; the α-amino acid content was elevated in fish kept at high density. In conclusion, plasma cortisol levels indicated an increasing stress level depending on the culture density, but significant changes in energy reserves did not occur in tissue (mainly liver and muscle glycogen and glucose reserves were significantly affected).