Growth performance of gilthead sea bream Sparus aurata in different osmotic conditions: Implications for osmoregulation and energy metabolism

The influence of three different environmental salinities (seawater, SW: 38 ppt salinity; brackish water, BW: 12 ppt; and low salinity water, LSW: 6 ppt salinity) on the growth, osmoregulation and metabolism of young gilthead sea bream (Sparus aurata L.) was studied over a period of 100 days. 480 inmature fish (20 g mean body weight) were randomly divided into six tanks of 2500 l (80 fish per tank) and maintained under three different salinities (38 ppt, 12 ppt and 6 ppt) in an open system. Every three weeks, 10 fish from each tank were anesthetized, weighed and lenghed. At the end of experiment, 10 fish from each tank were anesthetized, weighed and sampled for plasma, brain, gill and liver. Gill Na⁺, K⁺-ATPase activity, plasma osmolality, ions (sodium and chloride), glucose, lactate, protein and triglyceride, and hepatosomatic index were examined. In addition, levels of glycogen, lactate, ATP and activities of potential regulatory enzymes (hexokinase, pyruvate kinase, glycogen phosphorylase, and glucose 6-phosphate dehydrogenase) were assesed in liver, brain, and gill. BW-acclimated fish showed a better growth with respect to SW- or LSW-acclimated fish (12 > 38 > 6 ppt). The same relationship was observed for weight gain and specific growth rate. Osmoregulatory parameters in plasma (osmolality, Na⁺ and Cl⁻ levels) were similar in SW- and BW-acclimated fish but significantly higher than those of LSW-acclimated fish. Gill Na⁺, K⁺-ATPase activity showed lower values in intermediate salinity (6 > 38 > 12 ppt). No changes were observed in metabolic parameters analyzed in plasma, whereas only minor changes were observed in metabolic parameters of liver, gills and brain that could be correlated with the higher growth rates observed in fish acclimated to BW, which do not allow us to attribute the best growth rate observed at 12 ppt to lower metabolic rates in that salinity.     

Seawater adaptation and growth of post-smolt Atlantic salmon (Salmo salar) of wild and farmed strains

Atlantic salmon smolts (Salmo salar L.) of wild (Namsen) and farmed (AquaGen) strains were transferred to full-strength seawater (33‰) for 0 (initial control group), 0.5, 1, 2, 4, 8, 14, 30, 42 and 60 days at three different temperatures (4.3, 9.4 and 14.3 °C). Freshwater temperature in each tank was adjusted to test conditions 10 days prior to transfer. Physiological adaptation was monitored as changes in plasma growth hormone levels, gill Na⁺,K⁺-ATPase activity, plasma chloride levels and survival in seawater. Overall, smolts from the wild strain were better able to tolerate transfer to seawater than smolts from the hatchery strain. A delay in the osmotic disturbance and a prolonged period of osmotic stress were observed at the low temperature. Circulating GH levels increased transiently in all groups during the first 12–48 h in seawater and long-term GH levels were positively correlated with seawater temperature. Growth rates were influenced by temperature and strain, with the farmed smolts showing a higher growth than the wild smolts. Food Conversion Efficiency (FCE) was higher in smolts from the farmed strain, whereas no differences in daily food consumption were observed. Optimum temperature for FCE was calculated to be 10.5 °C, whereas the optimal temperature for growth in seawater was calculated to be 13.0 °C. We suggest that the observed differences in seawater tolerance, growth and food conversion probably are genetic and may reflect the fact that the hatchery fish have been bred for rapid growth for several generations.     

Plasma levels of vitamin D3 metabolites during parr-smolt transformation of Atlantic salmon Salmo salar L.

Plasma vitamin D and vitamin D metabolites were measured in Atlantic salmon parr during smoltification and after transfer to seawater. The fish were fed commercial feed for 5 months under natural light, and Na⁺/K⁺ ATPase was measured as an indicator of the smoltification status. No significant differences were recorded in the level of plasma vitamin D metabolites. However, a tendency of increasing plasma concentration of 25OHD₃ and also a temporary increase in plasma 1,25(OH)₂D₃ and 24,25(OH)₂D₃ were recorded prior to seawater transfer. The minor changes in plasma levels of the metabolites indicate a role of the vitamin D metabolites during parr–smolt transformation, although we do not know whether the increased levels are caused by increased synthesis of the metabolites or by decreased binding to receptors or decreased excretion.     

Rearing conditions to develop seawater tolerance in underyearling sockeye salmon smolt

ABSTRACT:   In order to confirm the conditions required to produce underyearling smolts, juvenile sockeye salmon Oncorhynchus nerka were reared under different photoperiod and temperature conditions. The level of smoltification was assessed by plasma sodium concentration 24 h after transfer to sea water (plasma-Na), gill Na⁺,K⁺-ATPase activity (gill-ATPase), and plasma thyroxine (plasma-T4) and growth hormone (plasma-GH) concentrations. Mean fork length of all groups reached more than approximately 8.5 cm at the end of the experiment. The fish that experienced an alteration in photoperiod from short day (8:16 h light–dark cycle) to long day (16:8 h light–dark cycle) showed significant elevation in gill-ATPase from 9 to 16 µmol inorganic phosphate per milligram protein per hour (µmol Pi/mg pro/h) and reduction in plasma-Na from 194 to 154 mM. This group displayed distinct pigmentation on their fin margins. However, in groups reared under constant short day and constant long day, changes in gill-ATPase and plasma-Na occurred vaguely at each different time and pigmentation on fin margins were not observed. The observed photoperiod-induced changes were greater at 7°C than at 11°C. Further, the alteration of the photoperiod induced two–threefold increases in plasma-GH and plasma-T4. These results suggest that lengthening of photoperiod from short to long daylight at 7°C causes smoltification accompanied by active secretion of growth hormone and thyroxine in underyearling sockeye salmon when the fish are larger than 8.5 cm in fork length.     

Effects of 17β-estradiol and 4-nonylphenol on smoltification and vitellogenesis in Atlantic salmon (Salmo salar)

The impact of 17-estradiol (E₂) and the putative estrogenic compound, 4-nonylphenol (4-NP), on smoltification and vitellogenesis in Atlantic salmon (Salmo salar) was investigated during a 30 day period starting late April. Three groups of fresh water (FW) fish (1 year old, mixed sexes, average weight 23 g) were injected once a week with 50 µg (0.18 µmol) 17-estradiol, 3 mg (13.6 µmol) 4-nonylphenol dissolved in peanut oil, or peanut oil alone as control. Every ten days, subgroups were challenged with 28 ppt seawater (SW) for 24h, and sampled together with subgroups of FW fish. Treatment effects were examined on vitellogenic and osmoregulatory parameters. E₂ and 4-NP treatment increased the total calcium and protein level in plasma and the hepatosomatic index of FW fish, both indicating an activated vitellogenesis in the liver. The presence of vitellogenin in the plasma of 4-NP- and E₂-treated groups was further indicated by the appearance of a high molecular weight vitellogenin band (550 kDa) in electropherograms produced by native gel electrophoresis. This band appeared in exactly the same position in both the E₂- and the 4-NP-treated groups but could not be detected in controls. During the 30 day treatment period, control fish approached the peak of smoltification, as indicated by a distinct silvery appearance, decreasing condition factor, increasing levels of gill Na⁺,K⁺-ATPase and improved hypoosmoregulatory performance in the SW-challenge test. Both E₂ and 4-NP treatments significantly inhibited the progress of smoltification, as judged by a significant reduction of gill Na⁺,K⁺-ATPase activity, relative -subunit Na⁺,K⁺-ATPase mRNA expression, gill chloride cell density and a poorer hypoosmoregulatory performance of treated fish. The impaired SW-tolerance of E₂- and 4-NP-treated fish was strongly correlated with a decreased gill Na⁺,K⁺-ATPase activity. Despite a difference in relative potency, the present study shows that 4-nonylphenol and 17-estradiol may have qualitatively similar inhibitory effects on smoltification and hypoosmoregulatory physiology of Atlantic salmon. Both 4-NP and E₂ activated the vitellogenic system, and the study supports the hypothesis that sexual maturation and smoltification are antagonistic, developmental phenomena in salmon. It is suggested that the presence of estrogenic compounds in the environment may negatively influence smoltification and migration in wild stocks of salmon.     

Enhanced hypoosmoregulatory response to growth hormone after cortisol treatment in immature rainbow trout,Salmo gairdneri

The growth-independent effect of ovine growth hormone (oGH) and oGH + cortisol treatment on seawater (SW) adaptation in immature rainbow trout, Salmo gairdneri was investigated. Fish were injected every second day with saline, 2.0 μg oGH/g or 2.0 μg oGH + 8.0 μg cortisol/g for a maximum of 8 injections in freshwater (FW). Subgroups were transferred to 28‰ SW after 4 or 8 injections, and changes in plasma Na⁺ and Cl⁻, muscle water content and gill Na⁺/K⁺-ATPase activity were measured. In both of the hormone-treated groups retained in FW, gill Na⁺/K⁺-ATPase activity and interlamellar chloride cell density increased. The effects were most pronounced in the oGH + cortisol group after 2 weeks of treatment. After transfer to SW most of the control fish died due to the osmotic stress, whereas in the hormone-treated groups, mortality was low and there was a positive correlation between pretransfer gill Na⁺/K⁺-ATPase and the ability to maintain ionic-osmotic homeostasis after SW transfer. After two weeks of oGH + cortisol treatment, gill Na⁺/K⁺-ATPase activity was maximal. In contrast, after SW transfer, Na⁺/K⁺-ATPase activity increased further in the oGH-treated group. This group regulated ionic-osmotic parameters less effectively than the oGH + cortisol-treated group. The data indicate that GH and cortisol are important hormones in the regulation of hypoosmoregulatory mechanisms in S. gairdneri.     

Functional morphology of chloride cells in killifish Fundulus heteroclitus, a euryhaline teleost with seawater preference

ABSTRACT:   Recent advances in the functional morphology of chloride cells in killifish Fundulus heteroclitus , a euryhaline teleost with seawater (SW) preference, were reviewed. Immunocytochemical detection of chloride cells with anti-Na⁺/K⁺-ATPase revealed transitional processes of the chloride cell distribution during early life stages. Chloride cells first appear in the yolk-sac membrane at an early embryonic stage, followed by their appearance in the body skin in the later embryonic stages. The principal site for the chloride cell distribution then shifts from the yolk-sac membrane and body skin during embryonic stages to the gills and opercular membrane in larval and later developmental stages. Morphologically distinct SW- and freshwater (FW)-type chloride cells were identified in adult killifish adapted to SW and FW, respectively. Both types of chloride cells are equally active in the two environments, but exert different ion-transporting functions. Following direct transfer of killifish from SW to FW, SW-type chloride cells were transformed into FW-type cells as a short-term response, followed by the promotion of chloride cell replacement as a long-term response. In killifish acclimated to low NaCl (0.1 mM) FW, intense immunoreactivity for vacuolar-type proton pump (V-ATPase) was detected in the basolateral membrane of FW-type chloride cells, whereas the immunoreactivity was much weaker in fish acclimated to FW with higher NaCl concentrations (1.0 and 10.0 mM). These results suggest the occurrence of active ion absorption in FW-type chloride cells and the involvement of V-ATPase in ion-absorbing mechanisms. In view of recent advances in this field, future chloride cell research should be considered in relation to the functional diversity of chloride cells.     

Effect of dietary vitamin C supplementation on the oxygen consumption, ammonia-N excretion and Na/K ATPase of Macrobrachium nipponense exposed to ambient ammonia

The 96-h LC₅₀ of ammonia-N and the effects of dietary vitamin C on oxygen consumption, ammonia-N excretion and Na⁺/K⁺ ATPase activity of Macrobrachium nipponense exposed to ambient ammonia were investigated. The results showed that the 96-h LC₅₀ of ammonia-N was 36.6 mg l⁻¹ for the freshwater prawn, M. nipponense, at pH 8.0. When prawns were exposed to high ambient ammonia-N concentrations, the oxygen consumption rate increased and ammonia excretion decreased. Dietary vitamin C supplementation led to higher oxygen consumption and lower ammonia excretion. Na⁺/K⁺ ATPase activity increased with increased ambient ammonia-N exposure in the range of 0–18.3 mg l⁻¹, and then was reduced at ambient ammonia-N 36.6 mg l⁻¹. Na⁺/K⁺ ATPase activities of prawns fed a vitamin C-supplemented diet were significantly lower than those of prawns fed a diet which was not supplemented with vitamin C.     

Physiology of seawater acclimation in the striped bass,Morone saxatilis (Walbaum)

Several experiments were performed to investigate the physiology of seawater acclimation in the striped bass, Morone saxatilis. Transfer of fish from fresh water (FW) to seawater (SW; 31–32 ppt) induced only a minimal disturbance of osmotic homeostasis. Ambient salinity did not affect plasma thyroxine, but plasma cortisol remained elevated for 24h after SW transfer. Gill and opercular membrane chloride cell density and Na⁺,K⁺-ATPase activity were relatively high and unaffected by salinity. Average chloride cell size, however, was slightly increased (16%) in SW-acclimated fish. Gill succinate dehydrogenase activity was higher in SW-acclimated fish than in FW fish. Kidney Na⁺, K⁺-ATPase activity was slightly lower (16%) in SW fish than in FW fish. Posterior intestinal Na⁺,K⁺-ATPase activity and water transport capacity (J_v) did not change upon SW transfer, whereas middle intestinal Na⁺,K⁺-ATPase activity increased 35% after transfer and was correlated with an increase in J_v (110%). As salinity induced only minor changes in the osmoregulatory organs examined, it is proposed that the intrinsic euryhalinity of the striped bass may be related to a high degree of “preparedness” for hypoosmoregulation that is uncommon among teleosts studied to data.     

Effect of hyperoxygenation and low water flow on the primary stress response and susceptibility of Atlantic salmon Salmo salar L. to experimental challenge with IPN virus

Intensive salmon smolt production normally includes reduced water flow and hyperoxygenation (added oxygen) of remaining water. There is little information on how different water quality parameters influence the fish health and the susceptibility to infectious diseases. The current experiment was carried out to evaluate if the combination of hyperoxygenation and reduced water flow (hyperoxic) can act as a chronic stressor to salmon in freshwater (FW) in such a way that it increases the susceptibility to IPN virus (IPNV) following seawater transfer. In FW, after 22 days of hyperoxic exposure plasma ion, TBARS and cortisol were measured. The cortisol levels were significantly (p = 0.011) higher in the hyperoxic group compared to controls maintained under normal oxygen saturation and water flow (normoxic), indicating chronic stress. Hyperoxygenation in FW caused decreased plasma [Cl⁻] compared to the normoxic group (p = 0.037), while [K⁺] tended to be higher in the hyperoxic group (p = 0.088). No significant differences were observed in plasma [Na⁺], total osmolality, TBARS or hematocrit, but there was a tendency towards a lower hct in the hyperoxic compared to the normoxic group. In SW the mortality was higher in the hyperoxic group challenged with IPNV (34%) compared to the normoxic group challenged with IPNV (20%) (p = 0.02), and no mortality was observed in the PBS injected fish. The challenged fish showed an overall increase in plasma cortisol day 8, 10, 12 and 14 post-challenge (p = 0.015, p = 0.000, p = 0.046 and p = 0.022 respectively). After SW transfer and challenge, plasma [K⁺] was elevated in both challenged groups, but no consistent trends were found for plasma [Cl⁻], [Na⁺] or total osmolality during the SW phase. There were no significant differences in the gene expression level of IFN 1, Mx and IL 1β prior to challenge, suggesting that the basic expression level of these genes were not affected by hyperoxygenation. IPNV was detected in kidney and pylorus, by immunohistochemistry, cell culture, and RT-PCR in head kidney. This experiment indicates that chronic stress induced by a combination of low water flow and hyperoxygenation increases the susceptibility to IPNV challenge.     

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.     

Immunolocalization of Na+, K+-ATPase-rich cells in the gill and urinary system of Persian sturgeon, Acipenser persicus, fry

Localization of Na⁺, K⁺-ATPase-rich cells in the gill and urinary system of Acipenser persicus fry was performed through immunofluorescence light microscopy using a mouse monoclonal antibody IgGα₅ raised against the α-subunit of chicken Na⁺, K⁺-ATPase. Different types of epithelia were clearly identified in the gill epithelium: epithelia of branchial arch, interbranchial septum, filament and lamellar epithelium. The Na⁺, K⁺-ATPase-rich cells were found in the epithelia of branchial arch, interbranchial septum, filament, interlamellar region and also in the lamellae. Histologically, the urinary system is divided into head kidney, trunk kidney and short caudal kidney. The head kidney is composed of the pronephric tubules and the haemopoietic tissues, while the trunk kidney is composed of a large number of glomeruli and convoluted nephrons. Each nephron consisted of a large glomerulus and tubules (neck, proximal, distal and collecting tubules) which connected to ureters. Posteriorly, ureters extended and joined together to form a small urinary bladder. In the urinary system, no specific fluorescence staining was observed in the glomerulus, neck segment and proximal tubules. The distal tubule cells and collecting tubule cells showed a strong immunostaining of Na⁺, K⁺-ATPase. Epithelia of ureters and urinary bladder also showed several isolated immunofluorescent cells. Immunofluorescent cells were rich in Na⁺, K⁺-ATPase enzyme which is very important for osmoregulation.     

Comparison of physiological smolt status in descending and nondescending wild brown trout (Salmo trutta) in a Danish stream

Abstract –  Downstream movement of a wild population of brown trout was examined in a small Danish stream in relation to morphological and physiological smolt status from March to May. Downstream movement was monitored in a Wolf-type trap covering all possible passage routes in the stream. Trout caught in the trap were classified as parr, pre-smolt or smolt based on morphological criteria and compared with trout randomly caught by electrofishing upstream of the trap. Representative gill samples from trap-caught and electrofished trout were analysed for gill Na⁺, K⁺-ATPase activity and used as a measure of physiological smolt status. Only a few parr occurred in the trap. Few pre-smolts occurred in the trap evenly in March and early April. In late April, pre-smolt movement peaked. By comparison, the main downstream movement of smolts occurred in distinct peaks through late March and April. The majority of fish caught in the trap were judged as pre-smolts or smolts based on morphological criteria's and they were characterised by relatively high gill Na⁺, K⁺-ATPase activity compared with trout judged as parr. Trout caught by electrofishing upstream the trap, were classified as parr, pre-smolts and smolts early in the season (March). During and after the main smolt-run in April the distribution of the remaining trout in the brook became skewed in favour of pre-smolt and parr. The study suggests that smolting trout initiate downstream movement once having reached a certain physiological smolt condition (judged by increased gill Na⁺, K⁺-ATPase activity).     

Influence of cortisol, growth hormone, insulin-like growth factor I and 3,3′,5-triiodo-l-thyronine on hypoosmoregulatory ability in the euryhaline teleost Fundulus heteroclitus

The capacity of cortisol, ovine growth hormone (oGH), recombinant bovine insulin-like growth factor I (rbIGF-I) and 3,3,5-triiodo-l-thyronine (T₃) to increase hypoosmoregulatory capacity in the euryhaline teleost Fundulus heteroclitus was examined. Fish acclimated to brackish water (BW, 10 ppt salinity) were injected with a single dose of hormone suspended in oil and transferred to seawater (SW, 35 ppt salinity) 10 days post-injection. Fish were sampled 24 h after transfer and plasma osmolality and gill Na⁺, K⁺-ATPase activity were examined. Transfer from BW to SW induced significantly increased plasma osmolality but not gill Na⁺, K⁺-ATPase activity. Cortisol (50 g g^–1 body weight) improved the ability to maintain plasma osmolality and to increase gill Na⁺, K⁺-ATPase activity. oGH (5 g g^–1 body weight) also increased hypoosmoregulatory ability and gill Na⁺, K⁺-ATPase activity. A cooperation between oGH and cortisol was observed in increasing hypoosmoregulatory ability but not in increasing gill Na⁺, K⁺-ATPase activity. rbIGF-I (0.5 g g^–1 body weight) alone was without effect in increasing salinity tolerance or gill Na⁺, K⁺-ATPase activity. rbIGF-I and oGH showed a positive interaction in increasing salinity tolerance, but not gill Na⁺, K⁺-ATPase activity. Treatment with T₃ (5 g g^–1 body weight) alone did not increase salinity tolerance or gill Na⁺, K⁺-ATPase activity, and there was no consistent significant interaction between cortisol and T₃ or between GH and T₃. The results confirm the classical role of cortisol as a seawater-adapting hormone and indicate an interaction between cortisol and the GH/IGF-I axis during seawater acclimation of Fundulus heteroclitus.     

Osmoregulatory actions of growth hormone in juvenile tilapia (Oreochromis niloticus)

Growth hormone (GH) effectively promotes seawater (SW) adaptation in salmonids, but little is known of its effect in tilapias. Experiments were performed to investigate the effects of recombinant eel GH (reGH) on osmoregulatory actions and ultrastructural features of gill chloride cells in juvenile tilapia, Oreochromis niloticus. Tilapia showed a markedly improved SW survival, when directly transferred from freshwater (FW) to 62.5% SW 24h after a single reGH injection (0.25 or 2.5 µg g^-1) or 3 reGH injections (0.25 µg g^-1 every other day). Plasma Na+ and Mg²⁺ levels were significantly reduced by reGH (0.25 and 2.5 µg g^-1) compared with saline injections; Ca²⁺ concentrations were reduced significantly by high dose of reGH (2.5 µg g^-1) after SW transfer. However, fish failed to survive more than 24h when directly transferred to 70% SW, although the fish treated with reGH could survive longer than the controls. When examined by electron microscopy, the chloride cells were identified as mitochondrion-rich and an extensive tubular system was induced by GH treatment. The results of the present study suggest that, similar to its effect on salmonids, GH also exerts acute osmoregulatory actions and enhances SW adaptation in juvenile tilapia. GH also stimulates the differentiation of chloride cells toward SW adaptation.     

Seasonal variation in osmoregulatory and metabolic parameters in earthen pond-cultured gilthead sea bream Sparus auratus

Seasonal variations in osmoregulatory and metabolic parameters were assessed in juvenile gilthead sea bream ( Sparus auratus ) cultured in earthen ponds under a natural photoperiod and temperature. Specimens were sampled, and the plasma, gill, kidney and liver were collected during winter 2005 and 2006 (January), spring 2005 (April), summer 2005 (July) and autumn 2005 (October). Plasma osmoregulatory parameters showed higher values in summer, while metabolic parameters presented different patterns of variations. Gill Na⁺,K⁺-ATPase activity decreased significantly in winter, while gill metabolite levels showed different patterns of variations among seasons. The enzymatic activities tested did not present a clear pattern of variation [(glutamate dehydrogenase (EC 1.4.1.2) (GDH) and hexokinase (EC 2.7.1.11) (HK)] or significant differences along seasons [glucose 6-phosphate dehydrogenase (EC 1.1.1.49)]. Kidney Na⁺,K⁺-ATPase activity decreased during summer and autumn. Different patterns of variation were observed in kidney metabolite levels while all the enzymatic activities assessed [lactate dehydrogenase-oxidase (EC 1.1.1.27) (LDH-O), HK and GDH] presented the highest values during summer. In the liver, metabolite levels and enzymatic activities did not show significant variations or present clear patterns of variation along different seasons. These results indicated seasonal variations in the osmoregulatory and metabolic parameters of different organs (blood, gill, kidney and liver) in earthen pond-cultured gilthead sea bream ( S. auratus ), which could be mainly attributed to seasonal changes in temperature.     

In vitro survival and the effect of water chemistry and oxidative chemical treatments on isolated gill amoebae from AGD-affected Atlantic salmon

Amoebic gill disease (AGD) of cultured salmonids in Tasmania is caused by the amphizoic parasitic amoeba Neoparamoeba pemaquidensis. The freshwater tolerance of amoebae isolated from the gills of AGD-affected salmon (predominantly N. pemaquidensis) was tested in vitro using a trypan blue exclusion assay. Amoebae exposed to water containing high concentrations of Ca²⁺ or Mg²⁺ (200 mg l⁻¹) showed high levels of survival up to 3 h of exposure. Exposure to water containing elevated Na⁺, choline chloride or water at different pH all had no significant survival of amoebae. Exposure of amoebae to different concentrations of chlorine dioxide, chloramine-T or hydrogen peroxide in artificially hard water demonstrated that chloramine-T and hydrogen peroxide were the most efficacious at killing amoebae in vitro. This work suggests that the hardness of freshwater may be an important factor for the survival of marine amoebae (predominantly N. pemaquidensis) on the gills of AGD-affected salmon and have significant implications with regard to the efficacy of freshwater bathing practices for the control of AGD on farms. Additionally, chloramine-T and hydrogen peroxide appear to be efficacious at killing marine gill amoebae in vitro and may be useful for the control of AGD in farmed Atlantic salmon.     

Effects of Low Salinities on Oxygen Consumption of Selected Euryhaline and Stenohaline Freshwater Fish

The amount of energy required for osmoregulation depends on the difference between internal and external concentrations of ions (Rao 1968; Farmer and Beamish 1969), changes in corticosteroid hormone levels (Morgan and Iwama 1996), glomerular filtration rates (Furspan et al. 1984), gill and kidney Na⁺, K⁺-ATPase activity (McCormick et al. 1989; Morgan and Iwama 1998), tissue permeability to water and ions, and gill ventilation, perfusion, and functional surface area (Rankin and Bolis 1984). Differences in the energetic cost of osmoregulation play a significant role in the difference in growth rate between seawater-and freshwater-adapted fish (Morgan and Iwama 1991; Ron et al. 1995; Wang et al. 1997). Oxygen consumption is an indirect indicator of metabolic rate in fish (Cech 1990) and can be used to determine effects of salinity changes on energy costs.     

Spawning of ocean pout (Macrozoarces americanus L.): evidence in favour of internal fertilization of eggs

Sperm physiology, in vivo artificial insemination and spawning of the ocean pout (Macrozoarces americanus L.), a marine bottom fish, were studied. Milt was collected from the reproductive tract of mature males by suction using a catheter. The uncontaminated milt, having a very low sperm concentration, contains highly motile spermatozoa and sperm motility was retained in vitro at 4 °C for at least 24 h in both seminal plasma and ovarian slime collected from the oviduct of pre-spawning females. Instead of activating sperm, dilution in sea water instantly immobilized the spermatozoa of ocean pout. Osmolarity and pH of ocean pout seminal plasma were in the ranges 365–406 mOsM and 7.2–7.5, respectively. A study of the ionic composition of ocean pout seminal plasma demonstrated the presence of various ions including Na⁺, K⁺, Ca²⁺, Mg²⁺, and Cl⁻, with a remarkably lower K⁺ concentration compared to that from other fish species. Since injections of milt containing motile sperm into the ovaries of pre-spawning females, which spawned in the absence of males, yielded fertilized ocean pout eggs, it is concluded that the ocean pout exhibits internal fertilization. The larvae hatched after 3 months of egg incubation in ambient sea water (9–10 °C). With proper timing of in vivo artificial insemination of mature females, fertilized ocean pout eggs can be obtained from fish reared in captivity.