Effects of environmental Ca2+ levels on branchial chloride cell morphology in freshwater-adapted killifish Fundulus heteroclitus

ABSTRACT: To examine the involvement of chloride cells in the uptake of Ca²⁺ in freshwater (FW) killifish, chloride cell morphology was compared in fish acclimated to different defined FW environments with Ca²⁺ concentrations of either 0.1 mM, 0.5 mM, or 2.5 mM. Numerous chloride cells were detected in whole-mount preparations of the gill filaments, which were stained with an antiserum specific for Na⁺, K⁺-ATPase. Chloride cells, located mostly on the afferent–vascular edge of the filaments, were larger at lower Ca²⁺ concentrations. Electron microscopic observations showed that in the 0.1 mM and 0.5 mM Ca²⁺ experimental groups, the apical membrane of chloride cells were flat or slightly projecting and equipped with numerous microvilli. In the 2.5 mM Ca²⁺ group, some chloride cells formed an apical pit, whereas other cells were similar to those observed in the 0.1 mM and 0.5 mM Ca²⁺ groups. Plasma osmolality decreased with decreasing ambient Ca²⁺ concentration, suggesting that environmental Ca²⁺ affects the permeability of the body surfaces. Gill Na⁺, K⁺-ATPase activity in the 0.1 mM and 0.5 mM Ca²⁺ groups were significantly higher than that in the 2.5 mM Ca²⁺ group, implying the involvement of the Na⁺–Ca²⁺ exchanger in Ca²⁺ uptake in the gills. These findings suggest that chloride cells function as the site for Ca²⁺ uptake in killifish acclimated to low Ca²⁺ environments.     

Effects of the probiotic, Lactobacillus acidophilus, on the growth performance, haematology parameters and immunoglobulin concentration in African Catfish (Clarias gariepinus, Burchell 1822) fingerling

This experiment was carried out to evaluate the effects of the probiotic, Lactobacillus acidophilus , on the growth performance, haematology parameters and immunoglobulin concentration in African catfish Clarias gariepinus fingerling. Two experimental diets were formulated to contain 35 g kg⁻¹ crude protein and 10 g kg⁻¹ lipids accordingly and fed three times daily for 12 weeks to 25 C. gariepinus fingerlings per fibreglass tank in 12 replicates each. The control diet was prepared with no probiotic supplementation whereas the second diet was prepared supplemented with a probiotic, L. acidophilus , containing about 3.01 × 10⁷ colonies/g of diet. The results show that growth performance [specific growth rate (SGR) and relative growth rate (RGR)], nutrient utilization [protein efficiency ratio (PER) and feed conversion ratio (FCR)] and survival were significantly ( P <0.05) higher in fish maintained on the probiotic-supplemented diet compared with those on the control diet. Haematology parameters (packed cell volume, haemoglobin, erythrocyte sedimentation rate, red blood cell and white blood cell, total serum protein, Ca²⁺, Mg²⁺, Cl⁻, glucose and cholesterol) and total immunoglobulin concentrations were also significantly better in fish fed the probiotic-supplemented diet than in the control. Although the water quality parameters monitored were better in the fish fed the probiotic-supplemented diet than in the control, the parameters were not significantly different ( P >0.05). From the results of this experiment, we conclude that L. acidophilus can be used as a probiotic agent in African catfish culture, to enhance fish health, survival and better feed efficiency and growth performance.     

Effects of Heavy Metals on the Hatching Rates of Brine Shrimp Artemia salina Cysts

The effects of heavy metals (Hg²⁺, Cd²⁺, Co²⁺, Cr³+, Pb²⁺, Cu²⁺, Fe²⁺. Ni²⁺, Zn²⁺, Mn⁷⁺, Mn⁴⁺ and Mn²⁺) on the relative hatching percentage of brine shrimp, Artemia salina were assessed. Artemia salina cysts (San Francisco Bay Brand) hatched in seawater containing various heavy metal concentrations. After 48 hours, the number of hatched nauplii were counted. A negative linear relationship between hatching rate of Ariemia salina cysts and heavy metal concentration occurred, except copper which conformed to a cubic model. The relative toxicities of heavy metals on Artemia salina cysts were as follows (Cu²⁺, most toxic; Mn²⁺, least toxic): Cu²⁺ > Zn²⁺ > Ni²⁺ > Co²⁺ > Mn⁷⁺ > Cr³⁺ > Cd²⁺ > Fe²⁺ > Hg²⁺ > Mn⁴⁺ > Pb²⁺ > Mn²⁺.     

Determining the optimum doses of Kurasan (ethoxiquinolin) and butylhydroxytoluol (BHT) in dry pellets: effect of both anti-oxidants on some haematological and condition parameters of rainbow trout, Salmo gairdneri Richardson

Abstract. Kurasan and BHT were tested at doses of 100, 200 and 400mg kg^-1 incorporated in dry pellets. The administration of these antioxidants did not influence the red blood count of rainbow trout, Salmo gairdneri Richardson, at body weights of 100 to 400g, reared in flow-through tanks at water temperatures of 7 to 13°C and in cage culture at water temperatures of 15 to 2O°C. In laboratory experiments at a water temperature of 14°C, the highest Kurasan dose increased the red blood cell (RBC) count insignificantly by 14% (1.06 vs 0.93 10¹²1^-1) after 80 days, haematocrit (PCV) was increased by 27% (0.410 vs 0.32511^-1), and haemoglobin (Hb) by 16% (72 vs 62gl^-1), This was seen in the cage culture experiment, but not in the experiments in the flow-through tanks. A trend of diminishing haematological parameters of the red blood count and total blood serum protein (TP) of the fish fed with BHT-stabilized diet was recorded only under laboratory conditions at the water temperature 9°C, The decline of RBC count in the experimental group with 0.04% BHT represented 14% (0.90 vs 1.05 10¹²1^-1), the decline in PCV amounted to 18% (0.328 vs 0.39811^-1), Hb to 17% (57 vs 69g 1^-1), and TP to 11% (39 vs 44g 1^-1).     

Purification and characterization of the amylase from a small abalone <Emphasis Type="Italic">Haliotis sieboldii</Emphasis>

ABSTRACT:   Amylase, with MW of 59 kDa, was purified from small abalone Haliotis sieboldii by ammonium sulfate fractionation, CM Sepharose Fast Flow and Sephacryl S-100 HR chromatographies. The optimal pH and temperature of purified amylase were 6.0 and 37°C, respectively. The purified enzyme was stable at pH 6.0–8.0 and low temperatures. It was activated by Ba²⁺, Mg²⁺, Ca²⁺, Co²⁺, Ni²⁺, Mn²⁺, K⁺, Ag⁺, Na⁺ and Li⁺, but completely or partially inhibited by Al³⁺, Cu²⁺, Cd²⁺, Hg²⁺ and Zn²⁺. EDTA could completely inhibit, while iodoacetamide, N-ethylmaleimide and urea partially inhibit the purified amylase. According to the digestion mode of various polysaccharides, the purified enzyme was considered to be an α-amylase.     

Total sulphur amino acid requirement of the Indian major carp, Labeo rohita (Hamilton)

The quantitative dietary sulphur amino acid requirement of the Indian major carp, Labeo rohita (Hamilton), was determined by conducting a growth study. The experimental diets contained 400 g crude protein kg⁻¹ from casein, gelatine and supplemental crystalline amino acids. Diets containing six graded levels of methionine (3.2, 6.5, 9.0, 11.5, 14 and 16.5 g kg⁻¹) with a constant level of cystine (1.4 g kg⁻¹) were formulated and fed to triplicate groups of Labeo rohita fingerlings twice a day to satiation for 60 days. The optimum dietary requirement for methionine was estimated using the break-point regression analysis at 11.5 g kg⁻¹ of diet or 28.8 g kg⁻¹ of dietary protein. Thus the total sulphur amino acid (Met + Cys) requirement was determined to be 12.9 g kg⁻¹ of diet or 32.3 g kg⁻¹ of protein. Higher survival, specific growth rate and food conversion efficiency values were observed for fish fed the diet containing optimum levels of sulphur amino acids.     

Binding characteristics of the Zn-binding membrane protein from common carp

ABSTRACT:    This study incorporated the 43 kDa Zn-binding membrane protein isolated from common carp into liposome. The specificity and strength of the binding of ⁶⁵Zn to the 43 kDa protein-liposomes, and the binding of the ⁶⁵Zn-labeled 43 kDa protein-liposomes to laminin were studied. It was found that ⁶⁵Zn was bound to the external side of the 43 kDa protein-liposomes. Specific binding of ⁶⁵Zn to the protein-liposomes was detected. The binding parameter of Zn to the protein was found to be: maximum binding site (N_max), 76.7 pmole/µg protein (approx. 3 mole of Zn²⁺/mole); and equilibrium dissociation constant (Kd), 0.19 µM. Of the cations introduced (Ca²⁺, Cd²⁺,Co²⁺, Cr²⁺, Cu²⁺, Fe²⁺, Hg²⁺, Mg²⁺, Mn²⁺, Ni²⁺, Pb²⁺), only Co²⁺ competed significantly with Zn. The protein-liposomes were also found to bind specifically to laminin with a N_max of 1.1 pmole/µg laminin, and Kd of 4.79 µM. No significant protein-liposome binding occurred to other extracellular matrix proteins (fibronectin, fibrinogen or vitronectin). Furthermore, the binding was specifically inhibited by the Arg-Gly-Asp (RGD) peptide or GRGDSPG, while two other analogs (GRGESPG and GRADSPG) were without effect.     

Effect of dietary protein levels on growth of Penaeus stylirostris juveniles

The shrimp Penaeus stylirostris is currently produced on a commercial scale in Tahiti and New Caledonia. Both super-intensive (80 animals m⁻²) and semi intensive (25 m⁻²) systems are promoted. Locally produced commercial feed contains 380–400 g kg⁻¹ crude protein (CP) without special consideration for environmental impact. The need for a 'low pollution' diet implies reconsideration of the optimum dietary protein level for this species. Under experimental conditions, six isoenergetic practical diets ranging from 270 g kg⁻¹ to 440 g kg⁻¹ CP were formulated and fed to satiation for 30 days to juvenile P. stylirostris ; average growth rates were between 5.5 and 7.5 g per month with survival rates > 90%.
The lowest protein levels 270–310 g kg⁻¹, gave significantly ( P < 0.05) poorer growth (5.5 g per 30 days) than was observed with 330–430 g kg⁻¹ CP; 330 g kg⁻¹ CP may be recommended, and as it is lower than levels in diets used currently, there is a possibility of reducing nitrogenous waste. In addition to growth response, protein efficiency ratio, protein productive value and food conversion (feed/gain) all supported a recommended dietary protein level of 330 g kg⁻¹. Future prospects for practical feeds with even lower CP levels are considered.     

Evaluation of practical diets containing different protein levels on gonad development of female redclaw crayfish Cherax quadricarinatus

The effect of five experimental diets with different crude protein content (220, 270, 330, 390 and 450 g kg⁻¹) on gonad development of female Cherax quadricarinatus was tested under laboratory conditions. After 70 days, a significant linear relationship indicated that higher concentrations of protein and carbohydrates in the hepatopancreas were produced as the dietary crude protein increased ( P  < 0.05). There were significant responses of the gonadosomatic index, hepatosomatic index, biochemical composition of the gonad (protein, lipids, carbohydrates and energy) and frequency of secondary vitellogenic oocytes to dietary protein level, as indicated by significant fits of the quadratic equation to the observed experimental data. The optimal response of the criteria parameters corresponded to levels of crude protein in the range 284–355 g kg⁻¹. Overall, 330 g kg⁻¹ crude protein with a protein : energy ratio of 15.6 mg kJ^–1 was considered the most adequate concentration of dietary protein for gonad development and biochemical composition in female redclaw crayfish.     

The Effects of Different Types of Stress on Blood Glucose in the Giant Tiger Prawn Penaeus monodon

Abstract— Juvenile Penaeus monodon prawns were exposed to a variety of putative stressom and blood glucose concentrations monitored as an index of metabolic stress. Prawns were held under controlled conditions and food deprived for 12–18 h before blood sampling. Juvenile prawns maintained in tanks at densities of 9, 23, 36 and 50 prawns/m² for 4 wk did not have significantly different baseline levels of blood glucose, with mean concentrations varying from 1.1 to 1.3 mmol/L. In contrast, as an index of overall condition, antennal length and rostral spine length were significantly degraded in the 36 and 50 prawns/m² groups compared to the 9 and 23 prawns/m² groups ( P < 0.05). Blood glucose levels were significantly elevated after prawns were exposed to air for 10 min and also increased Significantly when dissolved oxygen was decreased from 6.5 mg/L to 2.1 mg/L. ( P < 0.05). Gassing of water from pH 8.5 to 5.9 with carbon dioxide, with a corresponding decrease of dissolved oxygen from 6.6 to 5.9 mg/ L, induced a rapid and significant rise in blood glucose levels from 1.1 to 2.3 mmol/L. In contrast, a decrease of water pH from 8.3 to 5.9 with sulfuric acid, did not result in a significant change in blood glucose concentrations. These findings are discussed in relation to pond management for maintaining prawns in a low stress environment.     

Carbohydrate level in the diet of silver barb, Puntius gonionotus (Bleeker) fingerlings: effect on growth, nutrient utilization and whole body composition

Five iso-nitrogenous (300 g crude protein kg⁻¹ diet) semi-purified diets with graded levels of carbohydrate at 220 (D-1), 260 (D-2), 300 (D-3), 340 (D-4) and 380 (D-5) g kg⁻¹ diet were fed ad libitum to Puntius gonionotus fingerlings (average weight 0.59±0.01 g) in triplicate groups (20 fish replicate⁻¹) for a period of 90 days to determine the effect of the dietary carbohydrate level on the growth, nutrient utilization, digestibility, gut enzyme activity and whole-body composition of fish. Fifteen flow-through cement tanks of 100 L capacity with a flow rate of 0.5 L min⁻¹ were used for rearing the fish. The maximum weight gain, specific growth rate, protein efficiency ratio, RNA:DNA ratio, whole-body protein content, protease activity, protein and energy digestibility and minimum feed conversion ratio (FCR) were found in the D-2 group fed with 260 g carbohydrate kg⁻¹ diet. The highest protein and energy retention was also recorded in the same group. However, from the second-order polynomial regression analysis, the maximum growth and nutrient utilization of P. gonionotus fingerlings was 291.3–298.3 g carbohydrate kg⁻¹ diet at a dietary protein level of 300 g kg⁻¹ with a protein/energy (P/E) ratio of 20.58 −20.75 g protein MJ⁻¹.     

The Potential Use of Mangrove Forests as Nitrogen Sinks of Shrimp Aquaculture Pond Effluents: The Role of Denitrification

A generalized nitrogen budget was constructed to evaluate the potential role of mangrove sediments as a sink for dissolved inorganic nitrogen (DIN) in shrimp pond effluents. DIN concentrations were measured in pond effluents from three semi-intensive shrimp ponds along the Caribbean coast of Colombia between 1994–1995. Mean NH₄⁺ concentrations in the discharge water for all farms were significantly higher (67 × 12 μg/L) than in the adjacent estuaries (33 × 8 μg/L). Average NH₄⁺ concentrations in the pond discharge over all growout cycles were similar, representing an approximate doubling in relation to estuarine water concentrations. In contrast, NO₂^-+ NO₃^- concentrations were similar in both pond effluent and estuarine waters. Dissolved inorganic nitrogen loading of the ponds was similar. The estimated reduction of DIN in pond effluents by preliminary diversion of outflow to mangrove wetlands rather than directly to estuarine waters would be × 190 mg N/m² per d. Based on this nitrogen loss and depending upon the enrichment rate, between 0.04 to 0.12 ha of mangrove forest is required to completely remove the DIN load from effluents produced by a 1-ha pond.     

Preparation of heavy meromyosin from the autolyzed squid mantle muscle homogenate

ABSTRACT:   Incubation of squid mantle muscle homogenate caused a selective cleavage of myosin into heavy meromyosin (HMM) and light meromyosin (LMM). HMM was isolated from the incubated homogenate by using ammonium sulfate fractionation. The purified HMM retained two types of light chain components. Its Mg²⁺-ATPase activity with or without F-actin showed a Ca-sensitivity. HMM was cleaved into subfragment-1 and subfragment-2 upon chymotryptic digestion with or without Ca²⁺, possessing different light chain composition. Two types of light chain component were kept intact when digested in the presence of Ca²⁺. Ca²⁺ stabilized HMM especially in a bound form to F-actin.     

Role of sodium bicarbonate on the initiation of sperm motility in the Japanese eel

ABSTRACT:   In order to find out the role of sodium bicarbonate (NaHCO₃) on the initiation of sperm motility in the Japanese eel Anguilla japonica , interactions were investigated between NaHCO₃ and various reagents (K⁺ channel blocker 4-aminopyridine [4-AP], ammonium chloride [NH₄Cl], sodium acetate and calcium chloride [CaCl₂]) that could regulate internal factors (intracellular K⁺, intracellular pH [[pH]_i] and intracellular Ca²⁺) in sperm motility. Contradictory effects of NaHCO₃ were observed (i.e. an inhibitory effect when 4-AP was absent and a promoting effect when 4-AP was present). Sodium bicarbonate inhibited the initiation of sperm motility in the Japanese eel. However, NaHCO₃ restored the motility of immotile sperm that 4-AP inhibited. The inhibitory effect of NaHCO₃ disappeared with the addition of NH₄Cl, which raised [pH]_i, but the promoting effect was not affected by [pH]_i. Although NaHCO₃ recovered motility in the presence of 4-AP, this recovery was also observed with the addition of CaCl₂ instead of NaHCO₃. In the initiation of sperm motility in the Japanese eel, two roles for NaHCO₃ are suggested: an inhibitory role relating to the regulation of [pH]_i and a promoting role relating to the uptake of another initiation factor, which could be Ca²⁺.     

A dietary energy level of 14.6 MJ kg−1 and protein-to-energy ratio of 20.2 g MJ−1 results in best growth performance and nutrient accretion in silver barb Puntius gonionotus fingerlings

Five iso-nitrogenous (300 g protein kg⁻¹ diet) and iso-lipidic (80 g kg⁻¹ diet) semi-purified experimental diets with variable energy levels of 10.5 (D-1), 12.5 (D-2), 14.6 (D-3), 16.7 (D-4) and 18.8 (D-5) MJ kg⁻¹ diets were fed to Puntius gonionotus fingerlings (average weight 1.79 ± 0.02 g) in triplicate groups (15 healthy fishes per replicate) for a period of 90 days to assess the optimum dietary energy level and protein-to-energy ratio (P/E). Fifteen flow-through cement tanks of 100 L capacity with a flow rate of 0.5 L min⁻¹ were used for rearing the fish. Maximum specific growth rate, protein efficiency ratio, protein productive value, RNA : DNA ratio, whole body protein content, digestive enzyme activity and minimum feed conversion ratio was found in fish-fed diet D-3 with 14.6 MJ kg⁻¹ energy level. There were no improvements in all these parameters with the further rise in dietary energy level. Hence, it may be concluded that the optimum dietary gross energy level for maximum growth and nutrient utilization of silver barb is 14.6 MJ kg⁻¹ diet with a resultant P/E ratio of 20.2 g protein MJ⁻¹ diet, when the dietary protein and lipid are maintained at optimum requirement levels of 300 and 80 g kg⁻¹ diet, respectively, for this species.     

Piscivory in larval perch (Perca fluviatilis): mechanisms structuring larval roach (Rutilus rutilus) cohorts

Abstract – Perch ( Perca fluviatilis ) can act as a piscivore from larval stage VI (body size 10.3 mm) on newly hatched larval roach ( Rutilus rutilus ), bream ( Abramis brama ) and smaller siblings of its own cohort. Consumption rates at this stage were approx. 0.5 prey/perch*h at 21°C. Larval perch predation was strongly gap-limited, and the maximum size of roach consumed by perch (perch length interval 10.3–62.0 mm) under experimental conditions followed the linear regression, P_prey-max.=0.478*L_Pred.+1.829 ( r ²=0.99, P <0.001, n =12). Under experimental conditions, predatory 0⁺ perch substantially affected the size distributions of 0⁺ roach prey cohorts, since smaller prey individuals were predated more frequently than larger ones. In both unimodal and bimodal size distributions of prey roach, the distributions changed according to the maximum prey size consumed by the added predatory perch. Unimodal prey distributions were positively skewed when piscivorous perch were added compared with controls without predators. According to the size distributions of lake-living 0⁺ roach and 0⁺ perch and the relative size difference between prey and predator, the vulnerability of 0⁺ roach cohort to 0⁺ perch predation changed from June to September. Prey vulnerability was extremely sensitive to the relative size difference between predator and prey. Therefore differences in hatching time and growth rates between the two species will strongly influence the potential for predator-prey interactions. ^Note     

Changes in Plasma Cortisol, Glucose, and Selected Blood Properties in the Summer Flounder Paralichthys dentatus Associated with Sequential Movement to Three Experimental Conditions

To determine the changes in blood chemistry associated with sequential transfer of summer flounder Paralichthys dentatus (320–480 g), 300 hatchery-reared fish were moved to three different environmental conditions during a 20-d period. Fish were transferred in progression from a recirculating seawater system (22 ppt, 22.5 C) to a flow-thru seawater system (31 ppt, 20.0 C), to three small coastal net pens (33 ppt, 15.5 C), and finally to a large open ocean net pen (33 ppt, 16.0 C). For this study, eight random fish were captured at each progressive step (environmental condition), anesthetized (MS222), and bled from the caudal vein (2 mL). Transferred flounder were bled every 12 h for 48 h to collect plasma cortisol and glucose samples. Fish were bled 24 h after transport and every 3 d thereafter for osmolarity, hematocrit, hemoglobin concentration, mean corpuscular hemoglobin content, glucose, cortisol, and the electrolytes Cl^- Na⁺, K⁺ and Ca⁺. The most significant perturbations to blood chemistry (P < 0.05) occurred within 24 h of initial transfer from the recirculating to flow-thru seawater systems, suggesting an osmoregulatory rather than handling or transfer related stress. Osmolarity, electrolyte, and hematological parameters fluctuated and then recovered to stable levels by day 8 in the flow-thru seawater system. However, unlike the initial transfer, successive movement to the coastal and then the open ocean net pens produced transient increases in both plasma cortisol and glucose levels, suggesting a high level of stress associated with extended flounder handling and transfer.     

Effects of Temperature on Growth and Protein Assimilation in Juvenile Leader Prawns Penaeus monodon

Abstract.— The effects of temperature on growth and protein assimilation in leader prawns Penaeus monodon were evaluated. Growth of leader prawns was significantly ( P < 0.05) affected by temperature. The highest growth rate was 0.124 g/d at 32.0 C and optimum growth occurred between 27.5–32.0 C. Growth was described by an equation of the form: W _t = W_oe^kt ( R² = 0.99). The relationship between temperature and growth was described by a curvilinear equation of the form: Y = a + bX + cX² ( R² = 0.70). Apparent protein assimilation efficiency was not significantly ( P > 0.05) affected by temperature.     

Delay of the egg activation process in the Black Tiger Shrimp Penaeus monodon by manipulation of magnesium levels in spawning water

The aim of this study was to determine whether magnesium (Mg²⁺) in seawater is required for egg activation of the black tiger shrimp Penaeus monodon and whether manipulation of Mg²⁺ levels can be used to delay the process and thereby synchronize egg activation. Female P. monodon broodstock were allowed to spawn in artificial seawater containing Mg²⁺ at varying levels with respect to the normal (100%) level: 100%, 50%, 20% and 0%. Egg activation occurred normally at 100% Mg²⁺, incompletely at 50% and 20% Mg²⁺ levels and did not occur at all with 0% Mg²⁺. The fertilization rate with 100% Mg²⁺ was observed to be 83%, but fertilization failed to take place in all the other groups. The fertilization rate was restored from 0% to 76% following the 20% Mg²⁺ level treatment when Mg²⁺ levels returned to normal (100%) as soon as spawning was completed. This study suggests that the level of Mg²⁺ in seawater plays a vital role in P. monodon egg activation, and that commencement of this process could be delayed by manipulation of the Mg²⁺ level during and immediately after spawning.     

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).