Elasmobranch pericardial function. 1. Pericardial pressures are not always negative

Acute studies have led to the generalization that negative pericardial pressure is necessary for optimal cardiac function in elasmobranchs. We chronically instrumented horn sharks with pericardial catheters to test the hypothesis that ejection of pericardial fluid through the pericardioperitoneal canal (PPC) during routine handling could have accounted in part for previous measurements of exclusively negative pressures (–0.3 to –9.1 cm H₂O) in elasmobranchs. Maximum and minimum pericardial pressures measured immediately following routine handling (acute pressures) were more negative than those measured in resting horn sharks at intervals from 1 to 27 days following handling (chronic pressures). Chronic pericardial pulse pressure was less than acute. Entirely positive pericardial pressures were observed on occasion. Handling of chronically catheterized horn sharks resulted in ejection of 21 per cent (range=10–26, n=5) of the initial pericardial fluid volume through the PPC and reduced pericardial pressure. Operating pericardial fluid volume of horn sharks averaged 2.0 ml.kg^–1 (range=1.6–2.6, n=9). The PPC opened after 4.3±0.2 ml.kg^–1 (x±S.E.) of elasmobranch saline had been slowly infused into the pericardium, corresponding to an average pressure of 1.3±0.2 cm H₂O (n=10). The presence of the PPC plus a comparatively large pericardial fluid volume allows horn sharks to regulate pericardial pressure. Our analysis of pericardial pulse pressure, which can be an index of cardiac activity, suggests in contrast to previous studies that the elasmobranch heart can have relatively high stroke volumes at pericardial pressures near ambient. Thus, for venous return in resting or even moderately active elasmobranchs, it is more important that pericardial pressure be pulsatile than at a mean level which is negative.     

Performance of the heat of the hagfish,Eptatretus cirrhatus

The maximum power output of isolated perfused ventricles of the hafish (Eptatretus cirrhatus) averaged 0.367±0.031 mW g^–1 (n=9), considerably high than estimates for the heart of the Atlantic hagfish (Myxine glutinosa). Maximal minute volumes averaged 21.55±1.28 ml min^–1kg^–1, with a mean stroke volume of 0.71±0.14 ml kg^–1 body weight, values which are similar to those reported for many teleost and elasmobranch hearts.Ventricular output showed the characteristic dependence upon atrial filling pressure up to an optimum filling pressure ofc. 4 mm Hg. At output pressures exceeding 14 mm Hg the stroke volume and power output fell sharply. At these afterloads, the ventral aorta remained distended following semilunar valve closure and so the volume of fluid ejected on ventricular systole was reduced. There was little change in the frequency of the heart as either input or output pressures were varied.     

Elasmobranch pericardial function. 3. The pericardioperitoneal canal in the horn sharkHeterodontus francisci

The pericardial and peritoneal spaces of elasmobranch fishes are connected by the pericardioperitoneal canal (PPC), which allows pericardial fluid to escape when pressures exceed 0.1–0.3 kPA. Using the horn shark (Heterodontus francisci), we tested the hypothesis that the PPC functions to increase cardiac stroke volume by lowering pericardial pressure during activity. We also assessed the role of the PPC during coughing, feeding, or burst swimming and examined the effects of PPC occlusion. Increases in heart size were not prevented following augmented venous return in sharks with undisturbed or occluded PCP, evidence that argues that pericardial fluid loss through the PPC is a cause of increased cardiac stroke volume and not the result. Coughs, feeding, and burst swimming led to discharge of pericardial fluid. Chronic PPC occlusion resulted in an increased pericardial pressure, fluid volume, and frequency of coughing, and a decreased survival time compared to shams. Thus, in the horn shark the PPC likely compensates for constraints that may be imposed by the pericardium, provides a route for pericardial drainage, and regulates cardiac stroke volume during periods of activity.     

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.     

Effects of sublethal, acidic aluminum exposure on blood ions and metabolites, cardiac output, heart rate, and stroke volume of rainbow trout, Oncorhynchus mykiss

Doppler flow probes were fitted around the ventral aorta of rainbow trout, which were exposed to combinations of pH and aluminum (pH 5.1–6.2, Al 0–80 g l^–1) for 60 h. Fish accumulated Al at the gill and exhibited decreased blood Na⁺, Cl^–, and Ca²⁺ concentrations, and increased K⁺, hematocrit, hemoglobin, glucose, and lactate, indicating increasing ionoregulatory disturbance with increasing Al concentration. Fish exposed to ambient water (6.2) or low-pH (5.3) water without Al exhibited slight reductions in heart rates, as well as increased stroke volume, resulting in little variation in cardiac output. In the presence of Al (20 to 40 g l^–1) at low pH (5.1–5.3), fish increased their heart rate slightly and generally maintained their stroke volume, resulting in increased cardiac output in the first two days of exposure. At the highest Al concentration (80 g l^–1, pH 5.1), tachycardia was observed, concomitant with a decrease in stroke volume. The ionoregulatory imbalance and resulting increased blood viscosity explain these increases in heart rate rather than stroke volume in fish exposed to high concentrations of Al.     

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.     

Nitrogenous waste excretion and accumulation of urea and ammonia inChalcalburnus tarichi (Cyprinidae), endemic to the extremely alkaline Lake Van (Eastern Turkey)

The endemic, anadromous cyprinidChalcalburnus tarichi is the only fish species known to occur in alkaline Lake Van (Eastern Anatolia, Turkey). EightC. tarichi were maintained individually in Lake Van water (17 – 19°C; pH 9.8; 153 mEq·I^–1 total alkalinity; 22 total salinity) and tank water samples analyzed for 24 h in 2 to 4 h intervals. At zero time, < 1µM ammonia was present and urea was undetectable in the tank water; at 24 h, total ammonia and urea made up 114±32 and 35±25µM, respectively. Over the experimental period, ammonia-N and urea-N excretion averaged 1041±494 and 607±169moles·kg^–1 fish·h^–1, respectively. The extent of urea excretion was highly variable between specimens. Uric acid excretion was not detectable.Urea was present at high concentrations in all tissues and plasma (25 – 35moles·g^–1·ml^–1) of freshly caughtC. tarichi; total ammonia content of the tissues was by a factor of 1.9 (liver) to 3.0 (brain) lower. High arginase activity (2.4±0.2 U·min^–1·g^–1) was detected in the liver ofC. tarichi but ornithine carbamoylphosphate transferase, a key enzyme of the ornithine-urea-cycle, was absent. Ureagenesis is likely through degradation of arginine and/or uricolysis. High glutamine synthetase activity (11±0.6 U·min^–1·g^–1) and low ammonia content in brain suggest that, like other teleosts,C. tarichi has an efficient ammonia detoxification in the brain, but in no other tissue.Nitrogenous waste excretion at alkaline pH is discussed. The ability ofC. tarichi to excrete high levels of ammonia at extremely alkaline pH is unique among teleosts studied so far. The mechanism of ammonia excretion under Lake Van conditions remains to be elucidated.     

A new technique of feeding, repeated sampling of blood and continuous collection of urine in white sturgeon

A new technique combining oesophageal intubation, dorsal aorta cannulation, and urinary catheterization was developed to monitor concentration of nutrients in the blood circulation and their metabolites in the urine of 1–2 kg white sturgeon, Acipenser transmontanus. Three experiments were conducted to assess the technique based on: (1) quantitative delivery of nutrients; (2) stress levels post-operation; and (3) monitoring of nutrients in the blood and metabolites in the urine. In Experiment 1, recovery of intubated Cr₂O₃ was 105±5% (mean ± SEM, n=3) 3 h after intubation. In Experiment 2, plasma cortisol and glucose concentrations returned to respective basal levels of 8.2±1.8 ng ml^–1 and 74± mg dl^–1 (n=9) 48 h post-operation. In Experiment 3, sturgeon intubated with 1 g kg^–1 body weight of glucose or dextrin at 48 h post-operation showed a significantly different (p<0.05) peak plasma glucose level of 139±4 and 100±5 mg dl^–1 (n=5), respectively, at 4 h. Urinary glucose excretion was 1.8±0.9 mg kg^–1 h^–1 (n=5) 4–8 h after intubation with glucose. Our results show that the new technique allows quantitative delivery of nutrients, repeated sampling of blood, and continuous collection of urine in white sturgeon with minimum stress.     

The physiological response to capture and handling stress in the Atlantic sharpnose shark, Rhizoprionodon terraenovae

Secondary effects of capture and handling stress in the Atlantic sharpnose shark, Rhizoprionodon terraenovae, were investigated in this study. Twenty-four sharks were serially bled and changes in several hematological parameters were monitored over a 1-h time period, following capture by hook-and-line. Blood samples were obtained from each shark at 0, 15, 30, 45 and 60 min. All 0-min (initial) blood samples were obtained within 3 min of capture. Blood glucose (9.2–13.1 mmol l^–1), lactate (1.5–28.9 mmol l^–1), and plasma osmolality (871–929 mOsm kg^–1) all increased after capture, whereas blood pH (6.86–6.78) declined. Hematocrit values (initial = 25.1%) remained unchanged throughout the 1-h stress period. Due to the short amount of time it took to obtain the initial sample and the lack of a significant relationship between the initial time and the initial parameter levels, all initial samples are considered the best approximation of the predisturbance resting levels. The use of repeated measures in this study enables us to describe the dynamics of the secondary stress response in the Atlantic sharpnose shark.     

Oxygen delivery to the fish eye: blood flow in the pseudobranchial artery of rainbow trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>)

Similar to gas secretion into the swimbladder, some teleosts also exhibit ocular gas secretion to supply their thick and avascular retina with oxygen. The process in the eye, however, is complicated by the fact that the retina is supplied with arterial blood only indirectly through the pseudobranch, a reduced gill arch of still unknown function. Approaches for closer analysis of the physiological implications of this serial blood supply arrangement and the associated process of O₂ release include complete unilateral vascular isolation of eye and/or pseudobranch while maintaining normal blood flow characteristics for undisturbed operation of the counter-current rete mirabile.Blood flow velocity in the afferent pseudobranchial artery (aPA; essentially equal to ophthalmic artery flow) of rainbow trout was measured by application of ultrasonic Doppler techniques. Accurate determination of flow rate in the untouched vessel was facilitated by mounting the Doppler crystal with a custom-made holder on the bone of the jaw and raw signal analysis by Fast Fourier Transformation for improved signal/noise ratio. Normal unilateral aPA blood flow was 746 l min^–1 kg^–1 body weight, (1.6% of cardiac output), the tissue-weight-specific blood flow was 3.6 ml min^–1 g^–1 for pseudobranch and 2.3 ml min^–1 g^–1 for the retina. Only about 28% of the oxygen potentially available with maximal activation of the Root effect is actually consumed in pseudobranch and retina. This generous supply of blood and the implicit safety margin may serve minimization of pH disturbances of retinal tissues, but may also come suitable during unilateral occlusion of supplying arteries for continued oxygen supply to the retinae. During contralateral occlusion, flow through the ipsilateral aPA increased to 136%. According to the normally low O₂ extraction, a minimal flow of 36% through bilateral anastomosis and contralateral ophthalmic artery may maintain complete contralateral oxygen supply.     

Effects of purified microcystin-LR and cell extracts of Microcystis strains PCC 7813 and CYA 43 on cardiac function in brown trout (Salmo trutta) alevins

Sub-lethal cardiac responses of brown trout alevins (Salmo trutta L.) were determined in response to aqueous extracts of the cyanobacterium Microcystis strains PCC 7813 (microcystins detectable by HPLC) and CYA 43 (no microcystins detectable by HPLC) and to the purified cyanobacterial hepatotoxin, microcystin-LR (MC-LR) at concentrations of 5, 50 and 500 g microcystin-LR equivalents l^–1. Responses were determined using a flow chamber and video camera attached to a low power microscope. Heart rate in brown trout alevins was acutely sensitive to cyanobacterial extracts and significant increases were observed within 15–60 sec of exposure to aqueous extracts, although no change was observed on exposure to purified MC-LR. Stroke volume increased in all treatments at 50 and 500 g MC-LR equivalents l^–1, which may, at least in part, have been due to vasodilation of the yolk-sac blood vessels. Cardiac output increased significantly at all three concentrations of cyanobacterial cell extracts but not at the lowest concentration of MC-LR, although the rate increased at levels at/or above 50 g l^–1. Increased heart rate, stroke volume and cardiac output occurred at environmentally relevant microcystin concentrations of Microcystis PCC 7813 and CYA 43 aqueous extracts.     

Basal H2-receptor stimulated and pH-dependent acid secretion from an isolated stomach mucosa preparation of the cod,Gadus morhua,studied using a modified pH-static titration method

Gastric acid secretion from isolated cod stomach mucosa was measured using a pH-static titration method. A basal acid secretion rate (BASR) of 6.0±0.6 nEqH⁺min^–1cm^–1 was measured when using 0.9% NaCl as luminal solution. There was a dose-dependent increase in response to histamine between 0.12 and 0.20 M (EC₅₀=0.15 M), above which gastric acid secretion plateaued at 13.5±1.8 nEqH⁺min^–1cm^–1. Ranitidine, a H₂-receptor antagonist, completely blocked the stimulatory effect of histamine and reduced the BASR. The H₁-receptor antagonist, clemastine, did not inhibit the response to histamine. Acid secretion rates decreased significantly when the pH of the luminal side of the mucosa was lowered from pH 5.75 to pH 4.50, indicating that a negative feedback mechanism was operating. Histological staining showed that oxynticopeptic cells were uniformly distributed throughout the cardiac stomach.It is concluded that the acid secretion in the isolated stomach mucosa of cod can be measuredin vitro with a pH-static titration method. The method was used to demonstrate that the BASR is downregulated by a decrease in pH. Furthermore, we conclude that the histamine receptor in the cod stomach mucosa resembles the mammalian H₂-receptor and that histamine is secreted under basal conditions.     

Elasmobranch pericardial function 2. The influence of pericardial pressure on cardiac stroke volume in horn sharks and blue sharks

The importance of negative pericardial pressure to heart function in clasmobranchs has been questioned due to the discovery of positive pericardial pressures in healthy horn sharks (Heterodontus francisci). We therefore implanted electromagnetic flow probes on the ventral aorta of horn sharks and blue sharks (Prionace glauca) and assessed stroke volume and pericardial pressure as pericardial fluid volume (PFV) was varied to test the hypothesis that elasmobranchs are capable of maintaining a relatively large stroke volume at pericardial pressures near and above ambient. Stroke volume was maximum between zero and 25% maximum PFV (maximum PFV: the volume of pericardial fluid required to open the pericardioperitoneal canal), where pericardial pressure was most negative. At 50% maximum PFV (pericardial pressure near or slightly above ambient) stroke volume was 70% and 98% of its maximum in horn sharks and blue sharks, respectively. At a larger PFV, stroke volume declined drastically, reaching zero where both PFV and pericardial pressure were maximum. Thus, at a pericardial pressure apparently associated with resting or mild activity, stroke volume is a relatively large proportion of the apparent maximum. Increased circulatory demands associated with strenuous activity may induce ejection of pericardial fluid through the perieardioperitoneal canal, which then lowers pericardial pressure. The lowered pericardial pressure causes an increase in stroke volume, indicating that control is in part effected by changing pericardial pressure.     

A method for the quantification and optimization of hydrodynamics in culture tanks

A method was developed to quantify hydrodynamic mixing parameters, and to optimize the physical environmental conditions, in culture tanks. Improved mixing will result in better tank water quality, more efficient use of available volume by the culture animals (leading to optimal stocking densities and better feed management) and possibly reduced water pumping requirements. Experiments were conducted to determine the influence of a range of flow rates, residence times, water depths and stocking densities on hydrodynamics in juvenile turbot (Scophthalmus maximus (L.)) tanks. Decreases in water depth resulted in significant improvements in mixing and the efficiency with which the water was used, as indicated by reductions in dead volumes. A depth of less than 9.4 cm at a flow rate of 2 l min^–1 was expected to minimize dead volumes in the tank. This indicated that mixing was better in shallower tanks. Within the range 0–13 l min^–1, increased flow rate improved mixing at a constant depth of 9 cm (and water volume of 18.54 l) though increased flow rates greater than about 2.5 l min^–1 produced only small improvements in mixing. Within the range 0–50 fish per tank (equivalent to a mean stocking density of 0–1.84 kg m^–2), stocking density did not significantly influence mixing in tanks with a depth of 9 cm and flow rate of 2 l min^–1. Such depth reductions, for demersal species, may be a useful means to either decrease water use without reducing residence time, or alternatively to increase the flushing rate without increasing water use, at a given stocking density. The large changes in the efficiency with which the tanks were used, which were achieved with ease, indicates that attention to water mixing can give positive benefits to a wide range of land-based farm operators. Care must be taken when adjusting tank hydrodynamics, that water quality is maintained and that biological parameters such as stress levels, sunlight effects and feed management are optimal.     

Cathepsin L in eggs and larvae of perch Perca fluviatilis: variations with developmental stage and spawning period

This study describes changes in cathepsin L activity during early development in Eurasian perch (Perca fluviatilis) and determines to which extend a precocious proteolysis of the yolk reserves by activated cathepsin L could be involved in the seasonal decline of egg and larval quality. During embryogenesis and early larval stages, samples were collected for determination of fertilization and hatching rates, assays of cathepsin L activity, and evaluation of larval resistance to stress (osmotic shock and fasting). Cathepsin L activity increased significantly during the embryonic development, reaching a maximum of 177.1±12.3 nmol min^–1mg fresh weight^–1 on day 1 post-hatching and significantly decreasing on day 3. Activity increased significantly during the spawning season and individual variations between egg strands became very high during the second half of the spawning period. An inverse relationship was established between the cathepsin L activity in 7-day old eggs and the decrease of hatching rate. A negative exponential regression was calculated between the cathepsin L activity of eggs and the resistance of corresponding newly-hatched larvae to osmotic shock. Resistance to fasting was significantly reduced in larvae from eggs with cathepsin L activity higher or lower than 20 nmol min^–1 mg^–1 (lethal time₅₀=4.7±0.7 and 3.5±0.4 days, respectively). The involvement of cathepsin L in the degradation of yolk reserves and its potential consequences on the quality of perch eggs and larvae are discussed in relation to results reported in salmonid fishes.     

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.     

Circadian heart rate changes and light-dependence in the Mediterranean seabream Sparus aurata

Despite the abundant literature on circadian heart rate rhythms in fish, few studies have attempted to elucidate if such pattern is endogenous or if it is modulated by environmental cues. In this study, heart rates were continuously recorded in Sparus aurata acclimated to different light regimes (normal or shifted LD 12:12 cycle and a pulsed 0.75:0.75 LD cycle) to investigate the endogenous nature and the role of light on the circadian pattern of heart rate. Under LD 12:12, the daily pattern of heart rate showed higher values at night and lower rates during the day (average amplitude of the heart rate change of 10.6 ± 5.7 beats min^–1). The circadian pattern was maintained in animals under constant light but the amplitude was decreased (2.4 ± 1.1 beats min^–1). Light masks the expression of the rhythm because under a pulsed LD cycle, heart rate tracks the pulsed LD phases, increasing during the dark phase and decreasing during the light phase. Besides the ultradian pattern imposed, an underlying circadian rhythm was detected, which indicates that the rhythm is endogenous. However, the strong link between heart rate and light obtained under pulsed LD cycles suggest the timing mechanism to be mostly based on external light reception.     

The effect of dietary vitamin E on the pleopodal egg number of Astacus leptodactylus (Eschscholtz, 1823)

The effect of vitamin E on the pleopodal egg number of Astacus leptodactylus (Eschscholtz, 1823) was studied. Crayfish were fed 2% of their total wet weight daily with vitamin E supplemented diets and a control for 70 days. The vitamin E content of the control diet, diet 1, diet 2 and diet 3 were 20 mg kg^–1, 40 mg kg^–1, 80 mg kg^–1 and 160 mg kg^–1 respectively on a dry weight basis. Vitamin E levels of the control and experimental diets were analysed by High Performance Liquid Chromatography. Results showed that diets containing supplemental vitamin E were associated with an increase in the number of pleopodal eggs. The best result was obtained with diet 2 containing 80 mg kg^–1 supplemental vitamin E.     

Induction of Out-of-Season Spawning of Pikeperch, Sander Lucioperca (L.)

Pikeperch were induced to spawn 3 months prior to the natural spawning period through photothermal and hormonal stimulation. Females (five specimens in each group) were stimulated with injection of human chorionic gonadotropin (hCG) once (200 IU kg^–1), twice (200 IU kg^–1, second dose after 48 h–400 IU kg^–1) or three times (200 IU kg^–1, after 24 h–200 IU kg^–1 and after another 24 h–200 IU kg^–1). The control group was injected once with 0.9% NaCl. The males were stimulated with a single hormone dose of 200 IU kg^–1. Eggs were obtained from all the hormonally treated fish. None of the control group females, which were only stimulated photothermally, ovulated any eggs. The time of ovulation was 66–71 h following the first injection, and the eggs viability until the eyed stage (from 71.5 to 77.5%) did not depend on the number of hormone doses (P > 0.05). The out-of-season spawning method described in this paper could be used to provide pikeperch larvae for intensive culture systems (recirculating water systems) before natural spawning season and to produce larger-sized pikeperch fingerlings for stocking.     

Tissue lipid peroxidative responses in Atlantic salmon (Salmo salar L.) parr fed high levels of dietary copper and cadmium

Atlantic salmon (Salmo salar L.) parr were fed for one month on fish meal based diets supplemented with Cd (0, 0.7, or 204 mg Cd kg^–1 DW) or Cu (0, 34, or 691 mg Cu kg^–1 DW) to assess the effects of non-essential (Cd) and essential (Cu) dietary metals on lipid peroxidation and the oxidative defence system. Cadmium accumulated significantly in the liver, intestine, and kidney of 204 mg Cd kg^–1exposed fish compared to controls. Copper accumulated significantly in the intestine, kidney, and liver of fish exposed to 691 mg Cu kg^–1, and in the intestine of 34 mg Cu kg^–1 exposed fish. Tissue Cu accumulation significantly increased intestinal and hepatic lipid peroxidation (as seen from thiobarbituric acid reactive substances, TBARS, levels) and subsequently decreased intestinal -tocopherol levels and increased intestinal and hepatic selenium dependent glutathione peroxidase (SeGSH-Px) activity. Dietary Cd significantly reduced SeGSH-Px activity in the intestine and liver of 204 mg Cd kg^–1 exposed fish compared to controls. No significant increase in tissue TBARS or reduction of -tocopherol levels was observed in the intestine of fish exposed to dietary Cd, with exception of the highest exposure group (204 mg Cd kg^–1). Dietary Cu caused depletion of tissue Se and glutathione levels, however the reduced availability of GSH and Se did not seem to explain the differences in SeGSH-Px activity. Dietary Cu had a direct effect on lipid peroxidation at a relatively low concentration (34 mg Cu kg^–1). Cadmium indirectly affected tissue lipid peroxidation by damaging the oxidative defence system at the highest dietary concentration (204 mg Cd kg^–1).