Does urea reabsorption occur via the glucose pathway in the kidney of the freshwater rainbow trout?

This study tested the hypothesis that the renal reabsorption of urea occurs via the glucose transport pathway in the freshwater rainbow trout (Oncorhynchus mykiss). The relationship between glucose transport and urea transport was examined by experimentally elevating the rate of renal glucose reabsorption via infusion of the fish with exogenous glucose, and by inactivating the glucose transporters via the administration of phlorizin. Under all treatments, urea was reabsorbed against a concentration gradient, with plasma levels of urea being higher than urine levels. Glucose was almost completely reabsorbed (88%) whereas urea was reabsorbed less efficiently (47%) but to a greater extent than water (22%). Glucose and urea reabsorption were both found to be correlated with Na⁺ and Cl^– reabsorption, though the latter were 20 fold and 200–300 fold higher than glucose and urea transport rates, respectively. Glucose infusions greatly increased glucose reabsorption but urea reabsorption was unaffected. Phlorizin treatment completely blocked glucose reabsorption, but urea reabsorption was again unaffected. We conclude that there is no relationship between glucose and urea handling in the trout kidney, thus disproving the hypothesis.     

Antioxidative role of selenium against the toxic effect of heavy metals (Cd<Superscript>+2</Superscript>, Cr<Superscript>+3</Superscript>) on liver of rainbow trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis> Walbaum 1792)

The main purpose of this study is to discuss the effect of Cd⁺², Cr⁺³ and Se metals on biochemical parameters in liver tissue of Oncorhynchus mykiss. The rainbow trout were exposed to heavy metal stress (Cd⁺², Cr⁺³) at 2 ppm dosage. The present study was undertaken to determine the protective effect of selenium treatment at the same dosage (2 ppm) on some biochemical parameters. The activity of catalase (CAT), glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and the changes in levels of malondialdehyde (MDA) from biochemical parameters were determined in liver tissue of the fish groups exposed to heavy metals, especially for the selenium-applied groups. Results of this study showed that the activities of CAT, GSH-Px and SOD in the tissues of fish exposed to the stress of Cd⁺² and Cr⁺³ were significantly lower than the control groups (P < 0.05). Meanwhile, the closer values to the control groups were obtained in selenium-added groups (Cr⁺³ + Se⁺⁴, Cd⁺² + Se⁺⁴). For the level of MDA, the last production of lipid peroxidation showed increases (P < 0.05) in the groups exposed to the metal stress, whereas significant decreases were obtained in selenium-applied groups. The result of the statistical evaluation showed that the negative effects occurring in the biochemical parameters of the applied groups exposed to the toxicity of heavy metal were significantly eliminated (P < 0.05) as a result of selenium treatment.     

Cortisol-induced changes in oxygen consumption and ionic regulation in coastal cutthroat trout (Oncorhynchus clarki clarki) parr

The influence of cortisol on oxygen consumption and osmoregulatory variables was examined in coastal cutthroat trout (Oncorhynchus clarki clarki) parr kept in fresh water (FW) and transferred to seawater (SW). Intraperitoneal implants containing cortisol (50 g g^–1) in vegetable oil resulted in elevated plasma cortisol titres similar to those observed in fish following a 24h SW exposure. Cortisol treatment significantly increased the oxygen consumption and plasma glucose levels of trout in FW, consistent with the glucocorticoid role of cortisol. Cortisol treatment did not cause any changes in plasma ion concentrations or gill Na⁺,K⁺-ATPase activity in FW after 10 days. Cortisol-implanted fish exposed to SW for 24h showed slightly improved ion regulatory ability compare to non-implanted controls. The results of this study suggest that during SW transfer in juvenile salmonids, increases in cortisol may act as both a mineralocorticoid and a glucocorticoid, depending on the developmental state of the fish (e.g., smolt versus parr). Furthermore, the relative energetic costs of osmoregulation and that of the stress associated SW transfer cannot be discerned using whole-animal oxygen consumption rates.     

饥饿及再投喂处理对草鱼生长、葡萄糖代谢和转运蛋白1表达的影响

设置持续投喂组(C,持续投喂8周)、饥饿再投喂组(R,饥饿4周+再投喂4周)和持续饥饿组(S,饥饿8周)3个处理组,研究3种不同饥饿处理对草鱼(Ctenopharyngodon idellus)血清生化指标、糖原和糖代谢相关酶和葡萄糖转运蛋白1(GLUT1)的影响,同时在此实验基础上研究草鱼在急性高糖负荷胁迫下的糖耐受能力、糖代谢相关酶和GLUT1的变化规律,旨在阐明草鱼在饥饿及再投喂处理条件下的糖代谢特征。选取初重为(125.35±0.54)g的草鱼,饲养8周后以30 mg/100 g体重的剂量腹腔注射葡萄糖研究其糖耐受能力。结果显示,S组肝糖原和血清的血糖、甘油三酯含量均最低。饥饿处理对草鱼糖耐受能力影响显著,S组血糖含量在各时间点上显著低于其余两组(P0.05),肝糖原在6 h达到峰值;饥饿处理对草鱼肝脏糖代谢关键酶影响显著,饥饿处理(S组)诱使磷酸烯醇式丙酮酸激酶(PEPCK)活性上升但抑制丙酮酸激酶(PK)和果糖-6-磷酸激酶(PFK)的活性(P0.05),而饥饿再投喂(R组)后PEPCK、PK和PFK酶活性恢复到持续投喂(C组)处理水平。注射葡萄糖后S组肝脏GK酶活性增幅最大,PK酶活性呈上升趋势,而R组则呈先下降后上升的趋势;饥饿处理对草鱼肝脏和肌肉GLUT1表达影响显著,注射葡萄糖后,除R组肝脏组织外,其余各组草鱼肝脏和肌肉组织GLUT1表达量均呈先上升后下降的趋势,且S组肌肉GLUT1表达量在各个时间点上均高于其余两组(P0.05)。研究结果表明,在不同饥饿处理下,草鱼可通过消耗肝糖原和甘油三酯及降低肝脏糖酵解相关酶(PK和PFK)活性和促进糖异生PEPCK酶活性来应对饥饿胁迫,而饥饿处理可诱使GK和PK酶活性上升、促进糖原合成和激活GLUT1基因的表达和转运来缓解草鱼急性高糖负荷,从而提高其糖耐受能力。     

Effects of sulfide on K+ flux pathways in red blood cells of crucian carp and rainbow trout

The effect of sulfide on K⁺ influx pathways was measured in red blood cells (RBCs) of sulfide-sensitive rainbow trout (Oncorhynchus mykiss) and sulfide-tolerant crucian carp (Carassius carassius). In trout RBCs, maximal inhibition of Na⁺, K⁺-ATPase was attained at 10 mol l^–1 sulfide and amounted to 32% without being influenced by pH between 6.7 and 8.3. Ouabain-resistant K⁺ influx in the absence and presence of sulfide was insignificant at pH values between 6.7 and 7.7. At higher pH values ouabain-resistant K⁺ influx increased, but was inhibited to about 15% by 30 mol l^–1 sulfide. In RBCs of crucian carp neither Na⁺, K⁺-ATPase nor ouabain-resistant K⁺ influx were affected by sulfide concentrations up to 850 mol l^–1. Differences in sulfide-sensitivity of K⁺ influx between both species can be based upon different properties of the membrane transporter themselves. The reduced Na⁺, K⁺-ATPase activity in trout RBCs may also result from a slightly reduced (by 9%) ATP level after sulfide exposure. In addition, intracellular sulfide concentrations were higher in trout RBCs as compared to crucian carp. In trout, intracellular sulfide concentrations reached extracellular levels within 5 min of incubation whereas sulfide concentrations in crucian carp RBCs remained about 2-fold lower than extracellular concentrations. Although the physiological basis of sulfide-insensitive K⁺ influx in crucian carp RBCs is currently unknown it may contribute to the extremely high sulfide-tolerance of this species.     

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.     

Effect of ascorbic acid supplement in vitro onrainbow trout sperm viability

We evaluated motility and fertilizing ability of rainbow trout Oncorhynchus mykiss semen obtained from fish fed diets without ascorbic acid and a diet supplemented with 870 mg kg^{minus 1} of ascorbyl monophosphate. Semen was stored in vitro on ice (0 °C) during 14 days. The spermatozoa from the supplemented group had the highest motility and lowest decline in fertilizing ability after storage. Lack of a positive effect of exogenous vitamin C on semen in fish deficient in ascorbic acid (milt was supplemented with 50 mg l^–1 of ascorbic acid) suggests that the positive effect of ascorbic acid on semen quality is related to its long-term effects during spermatogenesis.     

Hepatic glucose utilization and lipogenesis of hybrid striped bass (Morone chrysops × Morone saxatilis) in response to dietary carbohydrate level and complexity

The influence of carbohydrate level and complexity on in vitro hepatic glucose utilization and lipogenesis were determined in hybrid striped bass, Morone chrysops ♀ × Morone saxatilis ♂. Six isocaloric, isonitrogenous diets containing glucose, maltose, or dextrin at two different levels (200 or 400 g kg^?1 diet) were fed to adult fish for 15 weeks. Liver explants were obtained at near‐maximum postprandial glycaemic response and incubated with radioactive labelled substrates. Glycogen synthesis from [¹⁴C]glucose was not different among treatments and was less than ¹⁴CO₂ formation. ¹⁴CO₂ production increased as a function of carbohydrate level but was unrelated to carbohydrate complexity. There was no detectable conversion of [¹⁴C]glucose to lactate for any treatment. Rates of de novo lipogenesis from [1‐¹⁴C]acetate were high in comparison to [U‐¹⁴C]glucose or [9,10‐³H]palmitate incorporation into liver lipids and differed in response to carbohydrate level and complexity. [9,10‐³H]palmitate esterification was an order of magnitude less than glycogen and CO₂ production but 4–10 times greater than [¹⁴C]glucose incorporation into liver lipids. Palmitate incorporation did not differ among treatments. Incorporation of [¹⁴C]glucose into liver lipids was higher in fish fed diets containing 400 g kg^?1 carbohydrate. These data support the idea that glucose is not a major oxidative substrate in hybrid striped bass and indicate that the level of soluble carbohydrate should be limited to 200 g kg^?1 diet or less for hybrid striped bass.     

Cortisol treatment improves the development of hypoosmoregulatory mechanisms in the euryhaline rainbow trout,Salmo gairdneri

The effect of cortisol on osmoregulatory parameters was studied in rainbow trout, (Salmo gairdneri), kept in freshwater (FW) and/or transferred to seawater (SW). Repeated injections of 20 μg cortisol/g fish stimulated gill and gut Na⁺/K⁺-ATPase activity and reduced plasma Na⁺ and Cl⁻ levels after 2 weeks of treatment in FW-adapted fish. Cortisol doses of 0.05 and 1.0 μg/g were without effect. Repeated injections of 10 μg cortisol/g stimulated gill Na⁺/K⁺-ATPase activity and reduced plasma Na+ and Cl⁻ levels in fish in FW, and significantly improved ion regulation after their transfer to 28SW. Higher doses of cortisol (10 and 20 μg/g) induced hyperglycemia, whereas low doses (0.05 and 1.0 μg/g were without effect or induced hypoglycemia. Plasma glucose levels decreased in cortisol-treated fish transferred to SW, whereas transient hyperglycemia was seen in the control fish.     

Effects of pH on contractility of rainbow trout (Oncorhynchus mykiss) intestinal muscle in vitro

Extreme acid-base imbalances in salmonids have been shown to impact on gastrointestinal motility. Changes in pH are known to affect cardiac and skeletal muscle in fish and mammalian intestinal smooth muscle preparations. This study was conducted to determine if the contractility of rainbow trout intestinal muscle was sensitive to pH fluctuations and to begin the investigation into the mechanisms by which pH affected contractility. Isolated duplicate or triplicate segments of rainbow trout intestine were suspended in organ baths containing modified Krebs-Henseleit solution. The pH was adjusted by varying the concentration of CO₂ aerating the solution and with HCl or lactic acid. Contractility was determined as response to the administration of 5-hydroxytryptamine (5-HT, serotonin), KCl, and transmural stimulation within the pH range of 6.3 to 8.5. Optimum pH for proximal segments was 7.85 while the range was much wider for distal segments. Decreasing the pH with CO₂ resulted in a greater inhibition of smooth muscle contractility than when the pH was decreased in 100% O₂ with either HCl or lactic acid, particularly when the tissues are stimulated electrically. This effect was ameliorated as the fish attained sexual maturity, although the mechanisms involved were not clear. The Cl^-/HCO ₃ ^- exchanger appeared to be involved in recovery from acidotic stress, particularly at pH levels below 6.5. The role played by the Na⁺/H⁺ exchanger is still unclear. Although contractility was inhibited in the presence of the amiloride analog EIPA (5-(N-ethyl-N-isopropyl)amiloride), whether the inhibition was due to blockage of ion transport or to cytosolic effects is yet to be determined.     

The role of glycogen phosphorylase in the regulation of glycogenolysis by insulin and glucagon in isolated eel (Anguilla rostrata) hepatocytes

The effects of porcine, scombroid, and salmon insulins, and bovine and anglerfish glucagons on glycogen depletion and glycogen phosphorylase (GPase) activities were examined in freshly isolated American eel (Anguilla rostrata) hepatocytes. Eel liver GPase in crude homogenates was activated (increase in % GPase a) by phosphorylating conditions and was rapidly inactivated (less than 1 h) when a phosphatase inhibitor (fluoride) was absent. Caffeine inhibits, and AMP activates, the b form of GPase consistent with their effects on rat liver GPase. Both mammalian and fish glucagons increased glucose production in eel hepatocytes, but had more ambiguous effects on glycogen levels and GPase activities. The magnitude of bovine glucagon effects were dependent on the initial glycogen content of the cells; only at glycogen concentrations less than approximately 70 μmoles.g⁻¹ did glucagon significantly increase % GPase a. Anglerfish glucagon significantly increased cyclic AMP (cAMP) concentrations by 90% at 10⁻⁷ M, but had no effects at 10⁻⁹ M and 10⁻⁸ M. Scombroid and salmon insulins maintained hepatocyte glycogen concentrations and decreased glucose production, with these effects more pronounced at low (10⁻⁹ to 10⁻⁸ M) rather than high (10⁻⁷ M) hormone concentrations. Porcine and salmon insulins decreased total GPase and % GPase a activities, and salmon insulin decreased CAMP levels, but only at 10⁻⁸ M (by 44%). Glycogen is, therefore, depleted by glucagon and maintained by insulin in freshly isolated American eel hepatocytes, and these changes are accomplished, at least in part, by changes in the activities of GPase. Changes in cAMP do not explain all of the observed hormone effects.     

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.     

Interrelationships between gill chloride cell morphology and calcium uptake in freshwater teleosts

The involvement of the freshwater fish gill chloride cells (CCs) in trans-branchial calcium uptake (JinCa²⁺) was investigated. This was accomplished by assessing the interspecific relationships between the apical surface area of CCs exposed to the external environment and JinCa²⁺. Three species of freshwater teleosts, the rainbow trout (Oncorhynchus mykiss), the American eel (Anguilla rostrata) and the brown bullhead catfish (Ictalurus nebulosus), were used. Chronic (ten-day) treatment with cortisol in each species was used as a tool to evoke variations in both JinCa²⁺ and gill CC morphology in order to assess intraspecific relationships between CC surface area and JinCa²⁺. The results of quantitative morphometry, based on analysis of scanning electron micrographs, demonstrated that catfish possessed the lowest fractional area of exposed CC (CCFA) on the gill filament epithelium (12,744 ± 2248 m²/mm²) and was followed, in increasing order, by American eel (21,355 ± 981 m²/mm²) and rainbow trout (149,928 ± 26,545 m²/mm²). With the exception of catfish, chronic treatment with cortisol caused significant increases in CCFA owing to proliferation of CCs and/or enlargement of individual CCs (eel only). The rates of JinCa²⁺ closely reflected the CC fractional area in each species. The results of correlation analysis revealed significant correlations between CC fractional area and JinCa²⁺ in trout and eel. Owing to the absence of an effect of cortisol treatment, there was no significant correlation in catfish because of insufficient variation in CC fractional area in this species. CC fractional area was significantly correlated with JinCa²⁺ among the three species examined. These results suggest that CC is involved in calcium uptake in freshwater teleosts and that both intra- and interspecific differences in the rates of calcium uptake can be accounted for by variability in the surface area of exposed CCs on the gill epithelia.     

Insulin-receptor binding in skeletal muscle of trout

Two hundred rainbow trout (Oncorhynchus mykiss) age 0 +, weight range 11.3 – 11.5 g, were distributed randomly in two groups and maintained for five weeks on either 10% dextrin, or 20% dextrin diet. The fish were sampled 3–5 h and 18–20 h after the last feeding and insulin binding to partially purified insulin receptors in white and red skeletal muscles and to liver plasma membranes was assessed. Plasma insulin, plasma glucose, and liver glycogen content were analyzed in the same fish.Fish fed a diet with higher carbohydrate content (HC) had elevated insulin and glucose levels in peripheral blood, but lower liver glycogen contents compared to the fish fed a diet with lower carbohydrate content (LC). No growth retardation was observed in the fish from HC group.Three to five hours after the last feeding, insulin-receptor binding in white skeletal muscles was higher in HC group of fish, mostly because of an increase in number of high affinity binding sites. Eighteen to twenty hours after the last feeding this difference disappeared. In contrast, the specific binding of insulin to the liver plasma membranes appeared to be lower in the HC group of fish. The lower insulin binding to the liver plasma membranes observed 3–5 h after feeding, could be attributed to the lower quantity of binding sites, while the same phenomenon 18 h after feeding was likely a result of affinity changes. We conclude that higher glycemic levels observed in trout fed a HC diet as compared to LC group of fish, are not a consequence of impaired binding of insulin to its receptors in skeletal muscles.Presented in part at the Annual Meeting of American Society of Zoologists, December 26–30, 1989, Boston, U.S.A. Supported by a grant (Personal investigador en el extranjero) of Ministerio de Educacion y Ciencia, (CYCIT) Spain to J.G. and by grants of the U.S. National Science Foundation (DCB 8615551 and DCB 8915935 to E.P.) and North Atlantic Treaty Organization (NATO) 0926/87 to C. Ottolenghi and E.P.     

Effects of dietary glucose and starch levels on the growth,haematological indices and hepatic hexokinase and glucokinase mRNA expression of juvenile mirror carp (Cyprinus carpio)

The effect of the different dietary carbohydrate types and levels on growth performance, haematological indices and hepatic hexokinase (HK) and glucokinase (GK) genes expression involved in control of glucose metabolism, was studied in juvenile mirror carp (Cyprinus carpio). Two carbohydrates (glucose and starch) diets with two levels (250 and 500 g kg^?1) were fed to triplicate groups of 35 fish for 60 days. The best weight gain rate and specific growth rate were observed in fish fed with 250 g kg^?1 glucose diet and 500 g kg^?1 starch diet (P < 0.05). Fish fed with 500 g kg^?1 glucose showed low feed utilization, with the highest food conversion ratio and the lowest protein efficiency ratio (P < 0.05). Hepatosomatic index was significantly higher in fish fed with glucose diets and the 500 g kg^?1 starch diet compared to 250 g kg^?1 starch. CHOL, HDL‐C and LDL‐C were significantly highest in fish fed with 500 g kg^?1 starch than all other diets (P < 0.05). Hepatic GK mRNA expression level and activity were positively related to glucose and starch levels (P < 0.05). Correlation analysis showed that hepatic glycogen concentration was increased by dietary carbohydrate content (P < 0.05). These results suggest that GK may play a major role in the postprandial glucose utilization in juvenile mirror carp.     

Recruitment and effects of Discocotyle sagittata (Monogenea) infection on farmed trout

Farmed rainbow trout Oncorhynchus mykiss and brown trout Salmo trutta were monitored over 3 years for infection with the blood-feeding gill fluke Discocotyle sagittata. Parasite transmission is seasonal: new infections take place during summer/autumn, and transmission is generally negligible during winter/spring. There are 2 sources of infection for naïve fish-of-the-year: limited invasion when fish are in the raceways by riverborne larvae originating external to the farm; and internally, within the farm, when 0+ fish are transferred to ponds previously occupied by older cohorts of infected fish. Thereafter, infection levels continue to increase in rainbow trout primarily through transmission within the farm. Prevalence rose to 100% in 1+ fish by the end of their second summer. In O. mykiss, mean abundance reached 194 worms/host for 1+ fish (up to 489 worms/host) and 160 worms/host for 2+ fish. By contrast, in S. trutta, parasite prevalence never exceeded 85% and, after the first year's invasions, infection levels decreased over time: in 1+ and 2+ brown trout, parasite mean abundance was < 4 (maximum 15) worms/host. We present evidence of the detrimental effects of D. sagittata on the host: high burdens are associated with pale gills, decreased body condition and host mortality. Parasite burdens become overdispersed during the warmer part of the year, as prevalence and mean abundance increase. However, the degree of parasite overdispersion decreases over winter; we cannot distinguish whether decreased aggregation is due to parasite losses from infected fish (including immune-mediated parasite mortality) or parasite-induced host mortality.     

花鲈ncc、nkcc基因在海、淡水适应中鳃组织的表达与定位分析

为探究ncc、nkcc基因在花鲈渗透调节中发挥的作用,实验通过全基因组鉴定、多重序列比对、系统进化树构建以及蛋白结构预测对花鲈ncc进行了鉴定及序列分析,利用实时荧光定量PCR (qRT-PCR)检测ncc和nkcc在海水、淡水花鲈鳃组织中的表达水平,利用原位杂交技术确定ncc2和nkcc1a在海水及淡水花鲈鳃中的表达位置。结果显示,从花鲈中鉴定出2个ncc基因,即ncc1和ncc2,其编码序列(CDS)长度分别为2 691和3 120bp,编码896和1 039个氨基酸,在进化上具有保守性。ncc2在淡水花鲈鳃组织中的表达量显著高于海水,而nkcc1a在海水花鲈鳃组织中的表达量显著高于淡水,ncc1、nkcc1b、nkcc2在海淡水中的表达量则无显著差异。淡水适应过程中花鲈鳃组织中的ncc2的表达量逐渐上调,而nkcc1a的表达量逐渐下调;海水适应过程则呈现相反的表达趋势。此外,原位杂交结果显示,ncc2和nkcc1a基因分别位于淡水与海水中鳃组织的相邻鳃小片间的鳃丝上皮。以上结果表明,ncc2和nkcc1a基因分别编码淡水及海水花鲈鳃中重要的Na⁺及Cl     

Lipid composition and microsomal ATPase activities in gills and kidneys of warm- and cold-acclimated sea bass (Dicentrarchus labrax L.)

The response to cold of gill and kidney membrane lipid composition and microsomal (Na⁺+K⁺)-ATPase, Na⁺-ATPase and Mg²⁺-ATPase activities in reared sea bass (Dicentrarchus labrax L.) was investigated. Fish acclimation was carried out according to the seasonal cycle from August to March. No cold-promoted increase in fatty acid unsaturation was shown in gill and kidney polar lipids and in total lipids of mitochondria and microsomes. In both tissues the (Na⁺+K⁺)-ATPase exhibited positive compensation for cold acclimation whereas the Na⁺-ATPase displayed negative compensation. The Mg²⁺-ATPase showed no compensation in the gills and positive compensation in the kidneys. During cold acclimation the break in the Arrhenius plot of the (Na⁺+K⁺)-ATPase decreased, whereas breaks of both the Na⁺-ATPase and the Mg²⁺-ATPase activities remained unchanged. The results indicate that the sea bass does not adopt membrane unsaturation as a cold-facing strategy. The cold-promoted enhancement of (Na⁺+K⁺)-ATPase activity in osmoregulatory tissues may be advantageous to maintain efficient osmoregulation under thermodynamically unfavourable conditions.     

Response of rainbow trout gill (Na++K+)-ATPase and chloride cells to T3 and NaCl administration

With the aim of comparing the effects of oral T₃ and NaCl administration on trout hypoosmoregulatory mechanisms, three groups of rainbow trout (Oncorhynchus mykiss Walbaum) held in freshwater (FW) were fed a basal diet (C), the same diet containing 8.83 ppm of 3,5,3-triiodo-L-thyronine (T₃) (T) or 10% (w/w) NaCl (N) respectively for 30 d. They were then transferred to brackish water (BW) for 22 d and fed on diet C. Gill (Na⁺+K⁺)-ATPase activity and its dependence on ATP, Na⁺ and pH, number of gill chloride cells (CC), serum T₃ level as well as fish growth, condition factor (K) and mortality were evaluated. During the FW phase, as compared to C trout, T trout showed a two fold higher serum T₃ level, had unchanged gill (Na⁺+K⁺)-ATPase activity and increased CC number, whereas N trout showed higher gill (Na⁺+K⁺)-ATPase activity and CC number. At the end of the experiment the enzyme activity was in the order T>N>C groups and all groups showed similar CC number. Both treatments changed the enzyme activation kinetics by ATP and Na⁺. A transient increase in K value occurred in N group during the period of salt administration. In BW, T and N groups had higher and lower survival than C group respectively. Other parameters were unaffected by the treatments. This trial suggests that T₃ administration promotes the development of hypoosmoregulatory mechanisms of trout but it leaves the (Na⁺+K⁺)-ATPase activity unaltered till the transfer to a hyperosmotic environment.     

Effects of sodium selenite on some biochemical and hematological parameters of rainbow trout (Oncorhynchus mykiss Walbaum, 1792) exposed to Pb2+ and Cu2+

This study was carried out to understand the preventive effect of selenium (Se⁴⁺) on heavy metal stress induced by lead and copper in rainbow trout (Oncorhynchus mykiss). Variation in glutathione peroxidase (Se-GSH-Px) and superoxide dismutase (SOD) activity, and in malondialdehyde (MDA) levels in liver, spleen, heart, and brain tissues of rainbow trout after 72 h of exposure to Pb²⁺ and Cu²⁺ were investigated in the presence and absence of Se⁴⁺. In the presence of Se⁴⁺, Se-GSH-Px activity and SOD activity were found to be higher and MDA levels were lower compared with in its absence. Hematological parameters were also determined and it has been observed that total leukocyte count (WBC), mean cell volume (MCV), and mean cell hemoglobin (MCH) were increased and erythrocyte number (RBC), hemoglobin (Hb), and hematocrit value (Hct; P < 0.05) were decreased in fish exposed to heavy metals in the absence of selenium. Selenium presence recovered hematological parameters to normal levels. In the light of our findings, it could be stated that Pb²⁺ and Cu²⁺ lead to dramatic changes in biochemical and hematological parameters and selenium caused these parameters to converge to control levels when it was administered concurrently with these heavy metals.