Glucose-stimulated lipolysis in rainbow trout,Oncorhynchus mykiss,liver

Rainbow trout, Oncorhynchus mykiss, were used to characterize further the influence of glucose on hepatic lipolysis. Liver was removed from fed fish, cut into 1 mm³ pieces and incubated for up to 5 h in Hanks medium containing either 2 mM, 5.5 mM, 10 mM, or 25 mM glucose. Glucose-stimulated lipolysis was indicated by tissue triacylglycerol (TG) lipase activity and by medium concentrations of glycerol and fatty acids (FA). Triacylglycerol lipase activity in liver pieces incubated in the presence of higher concentrations (25 mM) of glucose was significantly higher than that in liver pieces incubated in lower concentrations (2 mM) of glucose, rising from 0.075 ± 0.002 (mean ± SEM) nmol FA released/h/mg protein to 0.092 ± 0.004 units. Similarly, higher concentrations of glucose stimulated significantly more FA release and glycerol release from liver pieces than that stimulated by lower concentrations of glucose. Glycerol release from liver pieces incubated in the presence of 10 mM glucose and 25 mM glucose was ca. 2-fold to 2.8-fold, respectively, higher than that from liver pieces incubated in the presence of either 2 mM or 5.5 mM glucose. Fatty acid release from liver pieces incubated in the presence of 10 mM or 25 mM glucose was ca. 1.8-fold higher than that from liver pieces incubated in the presence of either 2 mM or 5.5 mM glucose. Notably, increased glycerol release was not accompanied by a parallel increase in FA. Fatty acid reesterification was more pronounced in liver pieces exposed to higher glucose (10 mM and 25 mM) than in liver pieces exposed to lower glucose (2 mM and 5.5 mM). ¹⁴C-incorporation studies indicated that glucose serves as a carbon source for reesterified FA in trout liver. The route of reesterification appears to be from glucose to glycerophosphate to phosphatidic acid to diacylglycerol to TG. Increasing concentrations of glucose did not affect glycerol kinase activity, indicating that glucose-stimulated lipolysis was not accompanied by increased glycerol recycling within the liver. These results suggest that glucose stimulates fatty acid reesterification and directly enhances net lipolysis in trout liver incubated in vitro.A part of this study was presented at the Annual Meeting of the American Society of Zoologists, December 26–30, 1991, Atlanta, GA.     

Melatonin, but not somatostatin-14 influences in vitro steroidogenesis by ovarian follicles of rainbow trout

Ovarian follicles taken from sexually maturing rainbow trout at the mid-vitellogenic stage of ovarian development were incubated in vitro in the presence or absence of melatonin or somatostatin-14 (SRIF-14) to determine whether there is evidence of a direct action of these factors on gonadal steroidogenesis in fishes. The steroidogenic capacity of the ovarian follicles was assessed by measuring testosterone (T) and 17-estradiol (E₂) release into the incubation medium, and by examining the steroid metabolites produced following incubation of follicles with radiolabelled steroid precursors.Melatonin appears to elicit a biphasic effect on steroidogenesis by in vitro rainbow trout ovarian follicles; at a concentration of 1 × 10^–3 M, melatonin stimulated basal T and E₂ production, but at a concentration of 1 × 10^–2 M there was an inhibition of basal and sGtH-stimulated T and E₂ Melatonin may act to reduce the activity of specific steroidogenic enzymes, since there was evidence of melatonin at 1 × 10^–2 M enhancing the accumulation of [³H]17-hydroxyprogesterone in the medium following incubation with [³H]pregnenolone, possibly suggesting the inhibition of C_17,20-lyase activity. In contrast, SRIF-14, used at concentrations of 1 × 10^–8 M and 1 × 10^–6 M, had no effect on basal or sGtH-stimulated E₂ or T production by ovarian follicles, incubated in vitro.     

Stress-induced changes in the affinity and abundance of cytosolic cortisol-binding sites in the liver of rainbow trout,Oncorhynchus mykiss (Walbaum), are not accompanied by changes in measurable nuclear binding

Plasma cortisol levels and the number (N_max) and affinity (K_d) of specific hepatic cortisol-binding sites were determined in rainbow trout subjected to chronic confinement stress for 14 days. Confinement significantly elevated plasma cortisol levels to 47.3 ± 13.5 ng ml^–1 within 24h and although levels declined to 8.0 ± 3.0 ng ml^–1 after 14 days, they were significantly higher throughout than levels in unstressed control fish (< 2.0 ng ml^–1). There was a 60% reduction in cytosolic N_max in stressed fish during the first 24h of confinement (35.8 ± 7.9 cf. 129.0 ± 15.2 fmol mg^–1 protein), a decline which was sustained at 7 days after the onset of stress but, although numbers of binding sites in the liver of stressed fish were still lower than in unstressed fish, the difference was no longer significant after 14 days of confinement. There was an accompanying significant rise in the K_d of cortisol binding in stressed fish during confinement, from 4.0 ± 0.6 nM at time 0 to 8.4 ± 0.8 nM after 24 h confinement. This increment in K_d was sustained at a level significantly higher than in control fish throughout the 14 day confinement period, despite marked reductions in cortisol levels and increases in N_max in stressed fish. Throughout the study, specific binding of cortisol could not be consistently detected in high-salt nuclear extracts from stressed or unstressed fish, suggesting either that high-affinity binding sites for cortisol were absent from these preparations, that receptors were present but unable to interact with ligand because they were occupied, or that receptors were present but not being extracted. These possibilities were investigated using a range of extraction procedures, by varying the temperature of incubation, by employing dexamethasone as ligand and by examining binding in purified, intact, nuclei. Estradiol was employed as a methodological control throughout and substantial amounts of specific estradiol binding were detected in all compartments and preparations. Specific cortisol-binding sites were detected in intact nuclei of both stressed and unstressed fish, at levels an order of magnitude lower than estradiol binding in the same preparations. These data demonstrate that activation of the pituitary-interrenal axis leads to significant changes in the nature of target-tissue binding of cortisol in rainbow trout, and reveal a clear difference in the subcellular distribution of binding-sites for estradiol and cortisol, which reflects the situation in mammalian tissues.     

Population ecology parameters and biomass of golden grey mullet (Liza aurata) in Iranian waters of the Caspian Sea

This paper examines the changes in the population ecology parameters and biomass of golden grey mullet (Liza aurata) in Iranian waters of the Caspian Sea from 1991 to 2005. For most years during this 14-year period, we estimated the age structure of the catch, length–weight relationship, von Bertalanffy growth parameters, condition factor, natural and fishing mortality and biomass. Growth parameters were estimated as L_∞ = 62.7 cm, K = 0.15 year⁻¹, t₀ = −0.23 year⁻¹. The instantaneous coefficient of natural mortality was estimated as 0.350 year⁻¹ and the instantaneous coefficient of fishing mortality varied during the 14-year period between 0.111 to 0.539 year⁻¹. Biomass estimates of golden grey mullet, from the biomass-based cohort analysis were increased from 13,527 mt in 1991–1992 to 23,992 mt in 2002–2003. In 2004–2005, it was estimated to be 23,658 mt. We concluded that at the present time, the stock of golden grey mullet is not being over-fished.     

Gluconeogenesis in trout (Oncorhynchus mykiss) white muscle: purification and characterization of fructose-1,6-bisphosphatase activity in vitro

Studies of the enzyme fructose-1,6-bisphosphatase (FBPase) of rainbow trout (Oncorhynchus mykiss) have been undertaken in order to illuminate aspects of skeletal muscle gluconeogenesis in these animals. Maximal activities in crude homogenates of several organs suggest that the liver possesses the greatest FBPase activity on a unit g^–1 tissue basis but that the white muscle, owing to its bulk, contributes substantially to whole body FBPase activity. Studies of fructose-6-phosphate-1-kinase (PFK) and FBPase in crude homogenates of several organs suggests an important role for intracellular pH in regulating the relative carbon flux through the FBPase/PFK locus in vivo. Furthermore, a three-step purification scheme is described for trout white muscle FBPase by which a stable and homogeneous (by SDS PAGE) enzyme preparation (isoelectric point = 7.2; molecular weight = 37.6 kd) was obtained. Kinetic studies of the purified enzyme were undertaken at 20°C under conditions reflective of "rest" and "exercise/recovery" intramuscular pH in vivo. Affinity for substrate (F-1,6-P₂) was increased (Km = 6.88 versus 2.44 mol 1-^–1 as was enzyme activity when pH was lowered from 7.0 to 6.5. Various inhibitor metabolites are identified including F-2,6-P₂ (mixed-type inhibitor, K_i = 0.201 mol 1^–1, pH 7.0) and AMP (non-competitive inhibitor, K_i = 0.438 mol 1^–1, pH 7.0). Inhibition by F-2,6-P₂ was strongly alleviated by a reduction in pH from 7.0 to 6.5 (I₅₀ increased from 0.14 to 0.32 mol 1^–1). AMP on the other hand was a more potent inhibitor at pH 6.5 but this inhibition was totally reversed under conditions of citrate, NH₄ ⁺ and AMP typical of muscle during recovery from exercise in vivo. In purified white muscle enzyme preparations, FBPase demonstrated maximal activity at pH 6.5 whereas the optimal pH of PFK was 7.0 or greater. Indeed, it appears from these in vitro data that regulation by metabolite levels as well as pH are required for net FBPase flux in vivo. It is concluded, therefore that trout white muscle FBPase demonstrates the potential to play an important enzymatic role in the control of intramuscular gluconeogenesis in these animals. The results are discussed in relation to present knowledge regarding the metabolic responses of trout white muscle to, and its subsequent recovery from, exhaustive exercise.     

Ionoregulatory and respiratory responses of brown trout,Salmo trutta,exposed to lethal and sublethal aluminium in acidic soft waters

Soft water acclimated (Ca²⁺ 0.02 mM; Na⁺ 0.03 mM; K⁺ 0.01 mM; pH 7.0), cannulated brown trout (Salmo trutta) were exposed to various pH and aluminium (Al) regimes (pH 7.0, pH 5.0, pH 5.0 plus Al: 50, 25, and 12.5 g l^–1) for up to 5 days in order to determine (i) the sublethal concentration of Al at pH 5.0 for this species (ii) their ionoregulatory and respiratory status. No mortality or physiological disturbances were evident at pH 7.0 or pH 5.0. All trout died within 48 h at pH 5.0 in the presence of Al at 50 g l^–1 and 67% died over the 5 day period at pH 5.0 in the presence of Al at 25 g l^–1. Fish at these lethal Al concentrations showed significant decreases in arterial blood oxygen content (C_aO₂) but no changes in plasma osmolarity or the concentrations of plasma Na⁺, K⁺ and Cl^–. Physiological disturbance was more marked at the 50 g l^–1 Al concentration. The surviving fish at 25 g l^–1 showed few signs of physiological recovery while continually exposed to this regime. No fish died during the exposure to water of pH 5.0 containing 12.5 g l^–1 Al, but physiological disturbance was still apparent. These sublethally-stressed trout showed a transient decline in the plasma concentrations of Na⁺ and Cl^–1. Although C_aO₂ decreased, recovery was evident. The data suggest that in the brown trout, environmental Al concentration is as important as pH and calcium concentration in determining the physiological status of the fish.     

NAD+-linked isocitrate dehydrogenase in fish tissues

NAD⁺-linked isocitrate dehydrogenase was found in the brain, heart, gills, kidney, liver and muscle of trout, and in the liver and muscle of eel. A complex homogenization buffer containing 1 mM ADP, 5 mM MgSO₄, 5 mM citrate and 40% glycerol is required for retrieval of significant amounts of stable enzyme. The highest activities were found in brain of trout and the lowest in white muscle of trout and eel. The enzyme was partially purified from frozen trout heart to a final activity of 0.04 M/min/mg protein, and the kinetic properties of this partially purified enzyme were studied. The enzyme requires either Mn²⁺ or Mg²⁺ for activity, higher activities being observed with Mn²⁺. Saturation kinetics for DL-isocitrate were sigmoidal, apparent S_0·5=8.2±0.6 mM and n_H=1.8±0.2, in the absence of ADP, changing to hyperbolic, apparent S_0·5=1.4±0.3 mM and n_H=1.0, with 1 mM ADP added. Citrate and Ca²⁺ were found to activate the enzyme to a small extent. NADH strongly inhibited the enzyme, I₅₀=3.7±0.5 M. ATP was also found to be an inhibitor, I₅₀=7.2±1.4 mM. These properties are consistent with the role of the enzyme as a major control site of the tricarboxylic acid cycle.     

Glycogenolytic action of glucagon-family peptides and epinephrine on catfish hepatocytes

Glycogenolytic effects of salmon and mammalian glucagons, salmon glucagon-like peptide (GLP) and epinephrine were studied on liver cells isolated from catfish (Ictalurus melas). In spring and summer, salmo-glucagon (3×10^–10 to 3×10^–8 M) was more effective than its mammalian counterpart in the stimulation of glucose release and cAMP synthesis in hepatocytes. GLP was less potent as compared to both glucagons. -amylase activity was not affected by the treatment with either glucagon-family peptides or epinephrine.The comparison of the glycogenolytic effects of salmon glucagon to those of epinephrine reveals a greater potency of the latter hormone in the stimulation of cAMP synthesis, glycogen-phosphorylase activity and glucose release. Glycogen content in the liver cells was equally depleted after treatment with both of the two hormones.     

Comparison of 3,5,3′-triiodo-L-thyronine and L-thyroxine absorption from the intestinal lumen of the fasted rainbow trout,Oncorhynchus mykiss

The absorptions of 3,5,3-triiodo-L-thyronine (T₃) and L-thyroxine (T₄) from the intestinal lumen of the rainbow trout were compared in vivo. Tracer doses of [¹²⁵I]T₄ (⁺T₄) or [¹²⁵I]T₃ (^*T₃) were injected through an anal cannula into the duodenum of trout fasted for 3 days at 12°C, and radioactivity was measured in blood and tissues at 4–48 h. ^*T₃ was removed more extensively than ^*T₄ from the intestinal lumen and more radioactivity was absorbed into the blood and tissues of u+T₃-injected trout than ^*T₄-injected trout. HPLC analysis showed that a high proportion of the radioactivity in the plasma, liver, kidney and intestinal lumen of ^*T₃-injected trout remained as the parent ^*T₃. However, in ^*T₄-injected trout most plasma radioactivity was in the form of ¹²⁵I^–, and by 24 h a high proportion of luminal radioactivity was ¹²⁵I^–. By 48 h, over 4% of the injected ^*T₃ and 1% of the injected ^*T₄ dose resided in the gall bladder, primarily as derivatives of ^*T₃ or ^*T₄. We conclude that T₃ is absorbed more effectively than T₄ from the intestinal lumen of fasted trout, indicating the potential for an enterohepatic T₃ cycle.     

Effects of Chronic Aluminum Exposure on Swimming and Cardiac Performance in Rainbow Trout, <Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>

Rainbow trout were exposed to 0–80 μg l⁻¹ aluminum (Al) at pH 5.2 in synthetic soft water, for up to 8 weeks. Trout were submitted to an incremental swimming test to quantify their aerobic swimming capacity (U_crit). After a simple, non-invasive cardiac surgery to install Doppler flow probes, their heart rate, cardiac output and stroke volume were measured while swimming at increasing water velocities. Fish exposed to Al accumulated significant amounts of Al at the gills (0–80 μg g⁻¹) and in their liver (5–60 μg g⁻¹) and had decreases in swimming capacity, ranging from 11 to 21%. Analysis of cardiac parameters during swimming revealed that increases in heart rate were used in trout exposed to the highest concentrations of Al to increase cardiac output, whereas control fish tended to increase cardiac output through increases in stroke volume.     

The conversion of plasma HCO 3 − to CO2 by rainbow trout red blood cells in vitro: adrenergic inhibition and the influence of oxygenation status

This study employed a recently developed radioisotopic assay (Wood and Perry 1991) to examine the inhibition, induced by catecholamines, of the conversion of plasma HCO ₃ ^– to CO₂ in acidotic trout blood, and the influence of oxygenation status on the response. Blood was incubated in vitro at P_{CO 2}= 2 torr, and 10^–6 M noradrenaline was employed as the adrenergic stimulus. In particular we investigated whether the inhibition of plasma HCO ₃ ^– conversion could be explained by a limited supply of H⁺s for the intracellular HCO ₃ ^– dehydration reaction because of competition by the adrenergically activated Na /H⁺ exchanger. Hypoxia (P_{O 2}= 15 torr) was employed as a tool to intensify this competition. Hypoxia raised RBC pHi, pHe, and plasma total CO₂ concentration (C_{CO 2}) by the Haldane effect, and increased the magnitude of Na⁺/H⁺ activation, expressed as the change in the transmembrane pH gradient (pHe-pHi). However hypoxia did not alter the inhibition of the conversion of plasma HCO ₃ ^– to CO₂ caused by noradrenaline. Hypoxia itself stimulated the RBC-mediated conversion of plasma HCO ₃ ^– to CO₂ by about 20% in the presence or absence of noradrenaline. The conversion rate was strongly correlated with pHe, pHe-pHi, and plasma C_{CO 2} in these experiments, but not with pHi. We conclude that adrenergically mediated inhibition in the conversion of plasma HCO ₃ ^– to CO₂ by trout RBCs is not due to competitive limitation on intracellular H⁺s, but rather to changes in the electrochemical gradient for HCO ₃ ^– entry and/or to CO₂ recycling from plasma to RBC. The deoxygenated condition helps to promote CO₂ excretion at the level of the RBC.     

Larval sea lamprey release two unique bile acids** to the water at a rate sufficient to produce detectable riverine pheromone plumes

The discovery that the olfactory system of the anadromous sea lamprey is extremely sensitive to two unique bile acids (petromyzonol sulfate [PS] and allocholic acid [ACA]) produced by stream-resident larval conspecifics has lead us to hypothesize that these compounds function as a migratory pheromone. Here, we test whether lamprey release these bile acids to the water in quantities sufficient for them to function as a long distance attractant. Five experiments were conducted. First, high performance liquid chromatography (HPLC) of liver extracts from all three life history stages of this species established that only larvae produce PS and ACA; parasites and maturing adults produced no identifiable bile acids. Large quantities of PS and ACA were found in larval gall bladders. Second, HPLC analyses of larval lamprey holding waters established that recently-fed larvae held in the laboratory release these bile acids to the water, with PS being released at a rate of approximately 16 ng h^–1 animal^–1 and ACA at 5 ng h^–1 animal^–1. Fasted animals released little bile acid. Third, an investigation of bile acid release routes demonstrated that larvae release bile acids primarily via their feces. Fourth, a study of the stability of ACA and PS in river water found both to have a half-life of a day. Finally, theoretical extrapolations using these data suggest that PS and ACA are present in picomolar concentrations in lamprey streams, a concentration within the detection range of adults. In conclusion, these data demonstrate for the first time in a fish that bile acid release rates and modes are adequate for these compounds to have pheromonal function.     

Epinephrine and norepinephrine elevate 5′-monodeiodinase activity in rainbow trout liver slices

The 5′-monodeiodinase (5′-MDA) activity was measured in liver slices that were incubated for 3 hours with epinephrine (E) or norepinephrine (NE) in order to examine the influence of these catecholamine hormones on the regulation of hepatic monodeiodination of thyroxine (T₄) in rainbow trout. Both E and NE induced a dose-dependent increase in 5′-MDA activity and in addition, E stimulated the release of T₃ into the medium. In liver slices taken from trout that had been treated with the β-adrenoceptor inhibitor propranolol, the response to both E and NE was attenuated. The findings provide evidence of an action of these catecholamine hormones on the peripheral regulation of T₃ production, and suggest that the control operatesvia the β-adrenoceptors. Corresponding author.     

The effects of experimental anaemia on CO2 excretionin vitro in rainbow trout,Oncorhynchus mykiss

The effects of severe experimental anaemia on red blood cell HCO₃ ^– dehydrationin vitro were examined in rainbow trout,Oncorhynchus mykiss. After 5 days of anaemia (haematocrit=4.9±1.1%) induced by intraperitoneal injection of phenylhydrazine hydrochloride, fish displayed elevated arterial CO₂ tensions (anaemic PaCO₂=3.19±0.42 torrvs. control PaCO₂=1.35±0.17 torr) and a significant acidosis (anaemic pHa=7.73±0.04vs. control pHa=7.99±0.04). However, after 15–20 days of anaemia (hct=6.6±0.8%) induced by blood withdrawal, the arterial CO₂ tension was significantly lower than the control value, suggesting that physiological adjustments occurred within this time period to compensate for the lowered haematocrit. Compensation probably did not involve alterations in ventilation, which was unaffected by 5 days of anaemia (anaemic ;w=786±187 ml min^–1 kg^–1 vs. control ;w=945±175 min^–1 kg^–1), based on indirect Fick principle measurements.Potential adaptations to longer term anaemia at the level of the red blood cells were investigated using a radioisotopic HCO₃ ^– dehydration assay. Owing to the difference in haematocrits, the HCO₃ ^– dehydration rate for blood from anaemic fish was significantly lower than that for control fish following equilibration at the same CO₂ tension. This difference was eliminated when HCO₃ ^– dehydration rates were measured on blood samples adjusted to the same haematocrit, a result which implies that the intrinsic rate of CO₂ excretion at the level of the red blood cell was not up-regulated during anaemia. The difference was also eliminated by equilibrating the blood samples with CO₂ tensions appropriate for the group from which the sample was obtained,i.e., PCO₂=1.4 torr for control samples and PCO₂=3.2 torr for anaemic samples; each at the appropriate haematocrit. It is concluded that the elevated PaCO₂ helps to reset CO₂ excretion to the control level, but that some additional physiological adjustment occurs to lower the PaCO₂ after 15–20 days of anaemia.     

Characterization of angiotensin-converting enzyme in the gills of rainbow trout,Salmo gairdneri (Richardson)

Several physical and chemical parameters of angiotensin-converting enzyme (ACE) were determined using a spectrophotometric assay of gill tissue homogenates from rainbow trout. This assay is based on the evolution of free hippuric acid via enzymatic cleavage of histidyl-leucine from the synthetic substrate hippuryl-l-histidyl-l-leucine (HHL). Piscine ACE exhibited enzymatic and kinetic properties similar to those reported for the partially purified mammalian enzyme. Proteolytic activity was both temperature and pH dependent and demonstrated hyperbolic kinetics with an apparent Km of 2.5 mM. Hydrolysis of HHL was activated by Cl^– at concentrations between 20 mM and 100 mM. Captopril (1 × 10^–6 M) and MK-422 (1 × 10^–6 M) blocked trout gill ACE activity, however, EDTA was inhibitory only at high concentrations (1 × 10^–3 M). These results demonstrate that trout ACE is functionally similar to mammalian ACE and that the spectro-photometric assay for ACE developed by Cushman and Cheung can be applied to analysis of converting enzyme activity in fish tissue homogenates.     

Quantification of presumptive Na+/H+ antiporters of the erythrocytes of trout and eel

The presumptive Na⁺/H⁺ exchange sites of trout and eel erythrocytes were quantified using amiloride-displaceable 5-(N-methyl-N-[³H]isobutyl)-amiloride (³H-MIA) equilibrium binding to further evaluate the mechanisms of i) hypoxia-mediated modifications in the trout erythrocyte -adrenergic signal transduction system and ii) the marked differences in the catecholamine responsiveness of this system between the trout and eel. MIA was a more potent inhibitor of both trout apparent erythrocyte proton extrusion (IC₅₀ = 20.1 ± 1.1 mol l^–1, N = 6) activity (as evaluated by measuring plasma pH changes after addition of catecholamine in vitro) and specific ³H-MIA binding (IC₅₀ = 257 ± 8.2 nmol l^–1, N = 3) than amiloride, which possessed a proton extrusion IC₅₀ of 26.1 ± 1.6 mol l^–1 (N = 6) and a binding IC₅₀ of 891 ± 113 nmol l^–1 (N = 3). The specific Na⁺ channel blocker phenamil was without effect on adrenergic proton extrusion activity or specific ³H-MIA binding. Trout erythrocytes suspended in Na⁺-free saline and maintained under normoxic conditions possessed 37,675 ± 6,678 (N = 6) amiloride-displaceable ³H-MIA binding sites per cell (B_max, presumptive Na⁺/H⁺ antiporters) with an apparent dissociation constant (K_D) of 244 ± 29 nmol l^–1 (N = 6). Acute hypoxia (PO₂ = 1.2 kPa; 30 min) did not affect the K_D, yet resulted in a 65% increase in the number of presumptive Na⁺/H⁺ antiporters. Normoxic eel erythrocytes, similarly suspended in Na⁺-free saline, possessed only 17,133 ± 3,716 presumptive Na⁺/H⁺ antiporters (N = 6), 45% of that of trout erythrocytes, with a similar K_D (246 ± 41 nmol l^–1, N = 6). These findings suggest that inter- and intra-specific differences in the responsiveness of the teleost erythrocyte -adrenergic signal transduction system can be explained, in part, by differences in the numbers of Na⁺/H⁺ exchange sites.     

Stocking density and cohort sampling effects on endocrine interactions in rainbow trout

Studies were conducted to explore the effect of cohort sampling and stocking density on the interactions between plasma growth hormone (GH), thyroid hormone and cortisol concentrations in rainbow trout. Depending on the experimental design, plasma GH concentrations were either suppressed, or elevated by sequential removal of cohorts from the holding aquarium. Since plasma cortisol concentrations consistently increased during cohort sampling, regardless of experimental design, it would appear that the apparent correlation (direct or inverse) is the result of concomitant changes, i.e. not necessarily a cause-effect relationship. Plasma GH concentrations of rainbow trout were not correlated with eviscerated body weight.Trout stocked at 150 kg m^–3 exhibited a significantly lower mean growth rate, hepatosomatic index, hepatic lipid reserve, plasma triiodo-l-thyronine (T₃) concentration andin vitro hepatic T₃ production than trout stocked at 60 kg m^–3. These observations are consistent with the former group being food deprived or ration restricted. Stocking density appeared to have no effect on plasma GH or cortisol concentrations, or on the pituitary-interrenal axis response to stressor challenge, or the thyroid tissue response to exogenous TSH challenge.     

Utilization of carotenoids from various sources by rainbow trout: muscle colour,carotenoid digestibility and retention

Rainbow trout (Oncorhynchus mykiss) with a mean (sd) weight of 120 (2) g were fed diets supplemented with astaxanthin extracted from the yeast Phaffia rhodozyma (OY1 = 50 mg carotenoids kg^–1 feed, OY2 = 100 mg carotenoids kg^–1 feed), astaxanthin (AX = 100 mg astaxanthin kg^–1 feed) and canthaxanthin (CX = 100 mg canthaxanthin kg^–1 feed) for 4 weeks. Muscle analyses at the end of the experiment indicated a significantly higher carotenoid concentration in the AX group, while CX and OY1 groups were similar in spite of the differences in dietary concentration. The measure of total muscle colour difference (E^* _ab) between initial samples and 4 week ones was higher for the AX fish group but showed no significant difference between OY1, OY2, and CX. The hue and the reflectance ratio (R₆₅₀:R₅₁₀) of fish muscle increased in proportion to carotenoid intake. Digestibility (ADC) of yeast astaxanthin in OY1 and OY2 groups was significantly higher than that in the AX group. Canthaxanthin ADC was about one sixth of that of astaxanthin (AX group). Carotenoid retention in the muscle, expressed as a percentage of carotenoid intake, was higher for the AX group than that recorded for OY1 and OY2. According to ADC, carotenoid retention showed a marked lower value for the CX group. Muscle retentions were similar for astaxanthins from both sources.     

Growth and physiological changes in scaled carp and blue tilapia under behavioral stress in mono- and polyculture rearing using a recirculated water system

Growth performance, carcass composition, liver and blood parameters ofscaled carp (C), Cyprinus carpio, and blue tilapia (T),Oreochromis aureus, reared for eight weeks in twomonoculture (100%C and 100%T) and two polyculture (60%C–40%T and40%C-60%T) conditions were investigated. In polyculture 40%C–60%T bothspecies achieved the highest levels of specific growth rate and the lowestlevels of food conversion ratio and carcass lipid content. In addition, theyexhibited the highest values of plasma pO₂ and pH and the lowestvalues of plasma pCO₂, cholesterol and albumin, although thedifferences among treatments were not significant in the case of tilapia.Tilapia showed significantly lower plasma Cl^– levels than underthe other conditions. Carp in monoculture and tilapia in polyculture60%C–40%T had the lowest levels of specific growth rate and significantlyhigher levels of liver lipids and plasma triglycerides than in the other groups.In addition, carp in monoculture exhibited a significantly higher haematocritthan in polyculture. No significant variations among treatments were observedconcerning plasma cortisol, glucose, osmolality, Na⁺, K⁺,HCO₃ ^– andHCO₃ ^–/H₂CO₃ in either species.The combination of scaled carp and blue tilapia, in which blue tilapia were themain species, proved to be the best for both species. It was suggested thatgrowth and physiological changes under mono- and polyculture rearing, in anintensive system, seem to be as a result of a different state of stress relatedto fish behavior.     

Rates of oxygen consumption and ammonia excretion of juvenile Eurasian perch Perca fluviatilis L.

The impact of feeding, fish size (body weight from 18.5 to 56.5 g) and water temperature (20 and 23 °C) on oxygen consumption (OC, mg O₂ kg^–1 h^–1) and ammonia excretion (AE, mg TAN kg^–1 h^–1) was studied in Eurasian perch held in recirculation systems. OC for both fed and feed-deprived (3 days) fish was higher at 23 °C (278.5 and 150.1 mg O₂ kg^–1 h^–1) than at 20 °C (249.3 and 135.0 mg O₂ kg^–1 h^–1; P < 0.01). AEs for both fed and feed-deprived fish were also significantly higher at 23 °C than at 20 °C (P < 0.001). Water temperature and fish size had a significant impact on the oxygen:feed ratio (OFR, kg O₂ kg^–1 feed fed day^–1) and ammonia:feed ratio (AFR, kg TAN kg^–1 feed fed day^–1; P < 0.001). Their average values at temperatures of 20 and 23 °C were 0.17 and 0.19 kg O₂ kg^–1 feed fed day^–1 and 0.009 and 0.011 kg TAN kg^–1 feed fed day^–1, respectively.