不同盐度对斜带石斑鱼幼鱼血清离子浓度和激素水平的影响及其与鳃MRCs渗透调节功能的关系

为探究不同盐度对斜带石斑鱼幼鱼血清离子浓度和激素水平的影响及其与鳃线粒体丰富细胞(MRCs)渗透调节功能关系,实验将暂养于盐度为30的水体中的斜带石斑鱼幼鱼直接转移至盐度分别为5、10、20和30(对照组)的水体中,于7和15 d分别检测血清Na~+、K~+、Cl~–浓度和血清皮质醇(COR)水平,并于第15天观察幼鱼鳃MRCs分布和结构的变化。结果显示,随盐度升高,幼鱼血清Na~+、Cl~–浓度显著上升,而K~+浓度无显著变化;COR水平在盐度5和10实验组显著高于盐度20和对照组;随盐度上升,鳃MRCs体积增大,数量增多,且盐度20和对照组鳃MRCs体积显著大于盐度5和10实验组;鳃MRCs表面存在一个特殊的顶膜结构,在不同盐度下呈现不同形态:盐度5和10实验组中鳃MRCs顶膜开口较大,且其表面存在大量的微绒,而盐度20实验组和对照组中鳃MRCs的顶膜向内深陷形成了一个顶隐窝,开口较小,且其表面没有微绒毛。研究表明,斜带石斑鱼幼鱼鳃MRCs在盐度5的水体中可以很好地发挥渗透调节功能,使幼鱼在盐度5的水体中存活。     

Non-linear release of Bohr protons with haemoglobin-oxygenation in the blood of two teleost fishes; carp (Cyprinus carpio) and tambaqui (Colossoma macropomum)

Some teleost fishes exhibit a non-linear release of H⁺ with haemoglobin oxygen saturation (SO₂) in whole blood that may be related to the Root effect and a low apparent cooperativity in oxygen binding. To further investigate this correlation, the relationship between red cell pH (pH_i) and SO₂ was evaluated in two teleost fishes, the carp (Cyprinus carpio) and tambaqui (Colossoma macropomum). Carp possess a relatively small Root effect within the physiological pH range, while that in tambaqui is relatively large. It was therefore hypothesized that both species would possess a non-linear release of H⁺ with SO₂ under in vivo conditions, but that the degree of non-linearity would be less pronounced in carp than in tambaqui. Red cell suspensions of tambaqui showed a marked non-linear relationship between pH_i and SO₂ at in vivo pH values (pH_e=7.66), where the majority of Bohr protons were released between 50 and 100% SO₂. In whole blood of carp, the relationship between pH_i and SO₂ was almost linear at normal resting extracellular pH values (pH_e=8.10) where Hills n ₅₀ (cooperativity) was 1.3. Under acidic conditions (pH_e=7.11), n ₅₀ decreased to 0.9 and the release of H⁺ with SO₂ became slightly non-linear. The reduction in pH in carp blood was associated with the onset of the Root effect, and the oxygen tension at half saturation (P₅₀) increased from 3.8 to 38.2 mmHg. The relationship between total CO₂ and SO₂ in carp whole blood changed from being almost linear at pH_e 8.1 to being non-linear at pH_e 7.1, consistent with the relationship between pH_i and SO₂. Thus, possession of a Root effect could be a prerequisite for a non-linear release of Bohr protons with oxygenation, but the expression in whole blood may depend upon a species-specific cooperativity threshold. The non-linear release of Bohr protons with oxygenation may be a general phenomenon in teleosts with important implications for gas transport and acid-base homeostasis.     

Survival, growth and osmolality of greenlip abalone Haliotis laevigata (Donovan 1808) when exposed to different ionic profiles of inland saline water

Two experiments over 21 days were conducted to assess the effect of different ionic profiles (mainly K⁺) of inland saline water (ISW) on the survival, growth, osmoregulation and moisture content of juvenile greenlip abalone Haliotis laevigata. In experiment I, the juveniles were cultured in five different water types: ocean water (OW) as a control, inland saline water (ISW or ISW K⁺₀), ISW fortified with 33%, 66% and 100% K⁺ as in OW (ISW K⁺₃₃, ISW K⁺₆₆ and ISW K⁺₁₀₀ respectively). In experiment II, five different culture media, 100% OW(control), 75% OW mixed with 25% ISW (ISW₂₅), 50% OW mixed with 50% ISW (ISW₅₀), 25% OW mixed with 75% ISW (ISW₇₅) and 100% ISW (ISW or ISW₁₀₀), were tested. Within 21 days, all of the abalone in ISW (ISW K⁺₀ and ISW₁₀₀; Na⁺/K⁺ ratio of 111 L^?1) died compared with 100% survival in OW. One‐hundred per cent K⁺ fortification of ISW increased the specific growth rate (SGR) and serum osmolality of the surviving juveniles. The 96 h acute LC₅₀ value was between ISW₅₀ and ISW₇₅. The increased proportion of ISW in OW resulted in significantly (P<0.05) lower survival, SGR and serum osmolality of juveniles. The moisture content of the juveniles was not influenced by any water type. The results suggested that abalone cannot survive in raw ISW, but survival can increase when it is fortified with K⁺ and ionic ratios of ISW are brought closer to OW by mixing it with OW. Furthermore, abalone survival is influenced by other ions in ISW besides K⁺.     

Reabsorption of urea by the kidney of the freshwater rainbow trout

In light of recent evidence that carrier-mediated transport of urea occurs in the mammalian kidney, this study examined the renal handling of urea in freshwater rainbow trout (Oncorhynchus mykiss). Fish were fitted with indwelling arterial and urinary bladder catheters for the measurement of plasma and urine composition (urea, Na⁺, Cl^-, glucose, H₂O), glomerular filtration rate ([³H]PEG-4000), and urine flow rate, thereby allowing quantification of tubular reabsorption rates. The fractional reabsorption of urea (72%) was greater than that of H₂O (50%) but less than that of Na⁺, Cl^-, or glucose (95–100%) and occurred against an apparent concentration gradient, suggesting active reabsorptive transport; [urea] in the urine was only 59% of that in blood plasma. When fish were infused with exogenous urea loads, these patterns remained largely unchanged, and urea reabsorption increased in direct proportion to the filtered urea load. There was no evidence for saturation of the reabsorptive transport mechanism at urea filtration rates up to 4-fold above the normal range, representing urea loading rates that proved toxic. Extra-renal excretion, presumably through the gills, increased markedly, almost keeping pace with urea loading. This evidence suggests that carrier-mediated reabsorptive transport of urea occurs in the kidney and that plasma urea levels are normally subject to tight homeostatic control in freshwater trout.     

Growth and survival of juvenile lined seahorse,Hippocampus erectus (Perry), at different stocking densities

The lined seahorse, Hippocampus erectus (Perry), is an important species in both medicinal and aquarium trades. The aim of this study was to evaluate the effects of stocking density (1, 3 and 5 individuals L⁻¹) on the growth performance and survival of the early-stage juvenile H. erectus. The height (HT), wet weight, weight gain (WG) and specific growth rate (SGR) were affected significantly by the stocking density during the 40-day study. The HT, WG and SGR of the seahorse at 1 and 3 juveniles L⁻¹ were significantly higher than that at 5 juveniles L⁻¹. The survival of juveniles at the three stocking densities was not significantly different at day 25 (90.3 ± 4.5%, 86.7 ± 4.2% and 86.2 ± 3.8% for 1, 3 and 5 juveniles L⁻¹ respectively), but was significantly different at day 40 (87.8 ± 3.9%, 69.6 ± 4.2% and 52.9 ± 2.8% for 1, 3 and 5 juveniles L⁻¹ respectively). For the early-stage juvenile H. erectus, we recommend a stocking density of 3 juveniles L⁻¹, but the density should be reduced to 1–2 juveniles L⁻¹ to avoid reduced and variable growth and high mortality after 25 days.     

The influence of culture density and enriched environments on the first stage culture of young cuttlefish, Sepia officinalis (Linnaeus, 1758)

The culture of Sepia officinalis hatchlings and juveniles at different densities and enriched environments was investigated. Experiments were conducted to determine effects of culture density and the use of a substrate on growth and survival. Experiment I studied the effect of three different densities (52, 515 and 1544 hatchlings m⁻²). Experiment II tested the effects of the enriched environment, using a sandy bottom with pvc shelters. Experiment III tested the effects of density on growth, survival, feeding rates and food conversions. Cuttlefish were fed live grass shrimp at rates of 20% body weight per day (BW d⁻¹). Grass shrimp (Palaemonetes varians) was supplied ad libitum as food in all experiments. In experiment I, growth was different between the three densities, with highest growth for density of 515 hatchlings m⁻². IGR was of 8.8, 9.6 and 9.2% BW d⁻¹ for the three densities tested, respectively. Both groups of experiment II had similar growth. IGR was of 10.1 and 9.7% BW d⁻¹ for enriched and non-enriched environments, respectively. Densities of 10, 45 and 120 juvenile m⁻² were used in experiment III. Significant differences in feeding rates were only found between densities of 10 and 120 cuttlefish m⁻² during the last week. Results indicate that culture of cuttlefish hatchlings could be done in a non-enriched environment, with densities not exceeding 500 hatchlings m⁻² and minimum bottom areas of about 600 cm². Densities of 120 juveniles m⁻² in a minimum area of about 1083 cm² should be considered for juveniles between 5 and 25 g. This revised version was published online in August 2006 with corrections to the Cover Date.     

Effect of dietary protein and energy levels on growth,oxygen consumption,haemolymph and digestive gland carbohydrates,nitrogen excretion and osmotic pressure of Litopenaeus vannamei (Boone) and L. setiferus (Linne) juveniles (Crustacea,Decapoda; Penaeidae)

The influence of protein and energy levels on growth rate, survival, pre- and post-prandial oxygen consumption, ammonia excretion, haemolymph glucose (HG), glycogen in digestive gland and osmotic pressure (OP) in white shrimp Litopenaeus vannamei (Boone) and L. setiferus (Linne) juveniles was studied. Diets containing a high-quality protein at a protein/energy (P/E) ratio of 16, 26, 31 and 36 mg kJ⁻¹ were fed at 20% of shrimp body weight of two sizes: < 1 g and > 1 g. Both species showed a optimum P/E ratio of 36 mg kJ⁻¹ (33–44% protein and 6–23% carbohydrate) in juveniles < 1 g. For shrimp > 1 g, L. setiferus showed a higher growth rate in the diet with 16 mg kJ⁻¹ (27% protein; 32% carbohydrate) and L. vannamei between 26 and 36 mg kJ⁻¹ (33–44% protein and 6–23% carbohydrate). In both experiments, the growth rate of L. vannamei was 2–3 times that observed in L. setiferus. Routine oxygen consumption and apparent heat increment (AHI) of L. setiferus juveniles was two times higher than that observed in L. vannamei juveniles, which could indicate that L. setiferus has a higher metabolic rate. The O/N ratio varied according to protein level, with higher values (O/N = 180) with a 16-mg kJ⁻¹ diet and lower values (O/N = 73) with a 36-mg kJ⁻¹ diet in L. setiferus juveniles. A similar variation in O/N ratio was obtained in L. vannamei fed with all diets with an interval between 22 and 50. An inverse relation between ammonia excretion and HG, and digestive gland glycogen (DGG) in relation to an increase in the P/E ratio indicate that both shrimp species are well adapted to use carbohydrates and/or proteins from their diet. The higher values of hyper-osmotic capacity (hyper-OC) were observed in L. setiferus < 1 g fed with 36 mg kJ⁻¹ and the lowest in L. vannamei < 1 g fed with 31 mg kJ⁻¹. Intermediate values of hyper-OC were observed in both species fed all diets indicating that osmotic factors of juveniles < 1 g of both species are more affected by the P/E ratio than juveniles > 1 g. All results showed that juveniles > 1 g of both species are less dependent of P/E ratio than juveniles < 1 g. Litopenaeus vannamei is a most tolerant shrimp species with a high capacity to use a wide range of dietary P/E ratios for growth, which may be due to its lower energy requirements. Litopenaeus setiferus showed a lower capacity to accept different P/E ratios but the optimum P/E ratio obtained with this species shows that L. setiferus accept diets with a high carbohydrate level as well. These results demonstrate that there are nutritional and physiological differences that explain the differences that have been observed when both species were cultured in commercial ponds.     

FOOD AND FEEDING STRATEGIES IN CULTURE OF LARVAL AND EARLY JUVENILE PURPLE-HINGE ROCK SCALLOPS,Hinnites multirugosus (GALE)

The purple-hinge rock scallop is receiving increasing interest as a candidate for commercial mariculture. Hatchery culture of juveniles is prerequisite since natural recruitment of juveniles is variable and not dependable as a source of stock. Success in carrying larvae through metamorphosis has been limited. Critical to larval culture is a sound understanding of optimal food and feeding strategies. Growth and survival were examined in early larvae fed six algal diets at four separate concentrations. The data clearly illustrate the superiority of both Isochrysis galbana and a Tahitian (T) strain of Isochrysis. Feed concentrations of 1 × 10⁵ cells/ml were found to be optimal for these species of Isochrysis. An I. galbana diet supported 80% survival through metamorphosis. The other algae examined included Mono-chrysis lutheri, Rhodomonas sp., Carteria pallida, and a 1:1:1 mixture of Rhodomonas, Monochrysis, and Isochrysis. All these diets supported 5–6% survival, and the growth was only slightly better than starvation controls. A similar study was done with early juveniles (1 mm dia). The diets included: I. galbana, M. lutheri, Thalassiosira pseudonana (3H), T-Isochrysis sp., and Phaedactylum tricornutum. As with the larvae, superior growth and survival occurred with diets of both species of Isochrysis. Cell concentrations between 1 × 10⁵ and 2 × 10⁵ cells/ml were optimal. The other diets supported minimal growth. In the feeding runs of both larvae and early juveniles, cell densities in excess of 5 × 10⁵ cells/ml were detrimental.     

Volume regulation in glutathione-treated brook trout (Salvelinus fontinalis) erythrocytes

Brook trout erythrocytes that were washed with and suspended in Ringer's solution with reduced glutathione (1.0 mM) maintained steady state cell volume for up to 24h, while those without the thiol-protective agent steadily shrank. Changes in cell volume (measured as packed cell volume, PCV) were evoked by acidic media (Ringer's at pH 6.8), hypoosmotic solutions (or both) and intracellular K⁺ and Cl⁻ concentrations were monitored over 4h. Acid-swollen cells failed to volume regulate or release K⁺ but had significantly elevated intracellular Cl⁻ Osmotically-swollen cells at pH 7.8 but not at pH 6.8 underwent regulatory volume decrease (RVD) and returned to initial levels in 2h, accompanied by release of K⁺ and Cl⁻ In contrast, osmotically-shrunken cells did not show regulatory volume increase. The regulatory volume decrease and concomitant K⁺ release were dependent on Cl⁻ implying a direct or indirect coupling of K⁺ to Cl⁻ transport in volume regulation. RVD was partially blocked by 4,4′-diisothiocyanatostilbene-2,2′-disulfonic acid (DIDS, 0.1 mM), an anion exchange blocker, but was unaffected by amiloride (1.0 mM) which blocks Na⁺/H⁺ exchange. Amiloride and DIDS prevented the swelling response to low pH but had no effect on control cells, suggesting involvement of Na⁺/H⁺ and Cl⁻/HCO₃ ⁻ exchanges in acid-induced cell swelling. Quinine (1.0 mM) a known blocker of K⁺ channels, exacerbated the osmotically-induced swelling but had little effect on the subsequent RVD and release of KCl. The results suggest that low extracellular pH inhibits neutral C⁻-dependent K⁺ release and the resultant regulatory volume decrease in osmotically-swollen cells.     

Applying biofloc technology in the culture of juvenile of Piaractus brachypomus (Cuvier, 1818): Effects on zootechnical performance and water quality

The aim of this study was to assess the zootechnical performance and water quality of a cachama blanca (Piaractus brachypomus) culture using biofloc technology (BFT) versus a system with daily water exchange (DWE). To do this, 180 juveniles (mean initial weight: 5.40 ± 0.19 g) were distributed in 12 circular plastic tanks with stocking densities of 20 or 40 individuals m³; then, they were cultured for 91 days. BFT treatments kept a C:N ratio approximately of 15:1. Temperature, pH and oxygen were monitored daily, while the other variables were measured weekly. Most productive variables were significantly influenced by both culture system and stocking density with significantly higher values of daily weight gain, total weight gain and total length for fish kept in DWE 20. However, only minor differences were observed within the BFT system. With the exception of the toxic nitrogen compounds (NH₄⁺ and NO₂^?), all the other water quality parameters were within the acceptable ranges for the cultivation of tropical fish. Microorganisms started to settle from the first week. A total of 23 genera were present, the most outstanding of which being seven genera of ciliates and three rotifers, rhizopods and chlorophytes. In conclusion, both systems BFT and DWE are useful for increasing the production of P. brachypomus in captivity. Additionally, the BFT system can potentially be applied for growing juveniles of this specie in regions with scarce water resources.     

Extension of nursery culture of Scylla serrata (Forsskål) juveniles in net cages and ponds

To address the preference of mud crab farmers for larger size Scylla serrata juveniles (5.0–10 g body weight or BW; 3.0–5.0 cm internal carapace width or ICW), a study was conducted to compare the growth and survival of crab juveniles (2.0–5.0 g BW; 1.0–3.0 cm ICW) produced a month after stocking of megalopae in net cages when reared further in net cages installed in earthen ponds or when stocked directly in earthen ponds. In a 3 × 2 factorial experiment, three stocking densities (1, 3 and 5 ind m⁻²), two types of rearing units (net cages or earthen pond) were used. Megalopae were grown to juvenile stage for 30 days in net cages set inside a 4000 m² brackishwater pond and fed brown mussel (Modiolus metcalfei). Crab juveniles were then transferred to either net cages (mesh size of 1.0 mm) or earthen ponds at three stocking densities. After 1 month, no interaction between stocking density and rearing unit was detected so data were pooled for each stocking density and rearing unit. There were no significant differences in the growth or survival rate of crab juveniles across stocking density treatments. Regardless of stocking density, survival in net cages was higher (77.11±6.62%) than in ponds (40.41±3.59%). Growth, however, was significantly higher for crab juveniles reared in earthen ponds. The range of mean BW of 10.5–16.0 g and an ICW of 3.78–4.33 cm obtained are within the size range preferred by mud crab operators for stocking grow‐out ponds.     

Nitrite toxicity to Litopenaeus schmitti (Burkenroad, 1936, Crustacea) at different salinity levels

Litopenaeus schmitti juveniles (total length 15 ± 0.4 cm) were exposed to different concentrations of nitrite using the static renewal method at different salinity levels (5‰, 20‰ and 35‰) at pH 8.0 and 20 °C. The 24, 48, 72 and 96 h LC50 values of nitrite in L. schmitti juveniles were 40.72, 32.63, 24.63, and 19.12 mg L⁻¹ at 5‰; 53.52, 38.60, 27.76, 25.55 mg L⁻¹ at 20‰; 54.32, 47.87, 41.67 and 38.88 mg L⁻¹ at 35‰ salinity. As the salinity decreased from 35‰ to 5‰, susceptibility to nitrite increased by 33.4%, 46.7%, 69.2% and 103.3%, after 24, 48, 72 and 96 h of exposure respectively. Furthermore, we found that exposure of shrimp to nitrite caused an increase in oxygen consumption by 137.3%, 99.2% and 81.4% and an increase in the ammonia excretion level by 112.5%, 87% and 64.3% with respect to the control with decreasing salinity levels.     

Observations on induced breeding of the Amazonian fish tambaqui, Colossoma macropomum (Cuvier) using CPE and HCG treatments

Trials on induced breeding of the native Amazonian fish tambaqui, Colossoma macropomum (Cuvier), using CPE and HCG treatments, were carried out during 1992-93 at the Fish Culture Station in Natal, Brazil. Final maturation, ovulation and spermiation of tambaqui broodstock, reared in earthen ponds, were induced by injecting heteroplastic carp pituitary gland extracts (CPE) and human chorionic gonadotrophin (HCG) separately. The female spawners received double injections and the males received a single dose. Females were tranquillized using quinaldine and their genital openings were sutured before the resolving injection was administered. Latent period between the resolving injection and ovulation varied from 6 to 11 h according to the prevailing water temperature. Milt was obtained by extrusion. Stripping and dry fertilization of the eggs produced viable larvae. Relative fecundity was estimated at 140 000 eggs kg^-1. Mean fertilization and hatching rates varied from 50% to 70% and from 60% to 80%, respectively.     

Preventing discoloration and lipid oxidation in dark muscle of yellowtail by feeding an extract prepared from mushroom (Flammulina velutipes) cultured medium

Groups of 0+ Atlantic salmon (Salmo salar) smolts were transferred to duplicate seawater tanks, and subjected to five different ration levels, 0% (starved), 25%, 50%, 75% or 100% (full fed). Waste feed was collected after each meal. After six weeks all groups were re-fed in excess. During the trial period body weight and length increased significantly in the 50, 75 and 100% groups, while no significant changes in body weight were observed in the 0% and 25% groups. A significant decrease in SGR was observed in the 0 and 25% groups during the first month in sea water. After re-feeding, SGR increased in all groups. All groups, except the previously starved group, showed peak SGR between weeks 6–8 and 8–12. Food restriction at 0% and 25% of full ration for a period of six weeks resulted in significant osmotic disturbances. After six weeks in sea water, plasma Cl⁻ levels were higher in the 0% group than in the other groups. Branchial Na⁺,K⁺-ATPase activity increased in all groups following exposure to seawater. Re-feeding caused a transient increase in branchial Na⁺,K⁺-ATPase activity after two weeks in the previously starved group, with a concurrent reduction in plasma Cl⁻ levels. Previous exposure to different ration levels significantly influenced growth rate and mean body size. Compensatory growth and partial size compensation was seen in the 0, 25 and 50% feed deprivation groups, whereas full size compensation was found in the 75% group.     

Eugenol as an efficacious anaesthetic for tambaqui, Colossoma macropomum (Cuvier)

Anaesthetics are important in fish culture to reduce handling stress and mortality. Eugenol is a promising anaesthetic because of its low cost, efficacy, safety margin for fish and lack of toxicity to humans. The goal of this study was to establish a protocol using eugenol as a fish anaesthetic for tambaqui Colossoma macropomum (Cuvier), and provide information for regulating authorities on establishing safety dosage protocols for its use. Juvenile and sub‐adult tambaqui were first individually exposed to doses of 35, 50, 65, 85, 100 or 135 mg L^?1 eugenol for 10 min. A second experiment examined the effect of the duration of exposure to eugenol on the time required for recovery and survival of tambaqui. A eugenol dose of 65 mg L^?1 was adequate to induce fish of both sizes into a surgical anaesthetic state, and recovery time was similar for dosages up to 100 mg L^?1. Exposure to the ideal dose (65 mg L^?1) for up to 30 min did not cause fish mortality. Fish blood glucose values were similar for all the tested eugenol doses as well as with the benzocaine control. The results show that eugenol is an efficient and safe anaesthetic for tambaqui.     

Immunolocalization of chloride transporters to gill epithelia of euryhaline teleosts with opposite salinity-induced Na<Superscript>+</Superscript>/K<Superscript>+</Superscript>-ATPase responses

Opposite patterns of branchial Na⁺/K⁺-ATPase (NKA) responses were found in euryhaline milkfish (Chanos chanos) and pufferfish (Tetraodon nigroviridis) upon salinity challenge. Because the electrochemical gradient established by NKA is thought to be the driving force for transcellular Cl⁻ transport in fish gills, the aim of this study was to explore whether the differential patterns of NKA responses found in milkfish and pufferfish would lead to distinct distribution of Cl⁻ transporters in their gill epithelial cells indicating different Cl⁻ transport mechanisms. In this study, immunolocalization of various Cl⁻ transport proteins, including Na⁺/K⁺/2Cl⁻ cotransporter (NKCC), cystic fibrosis transmembrane conductance regulator (CFTR), anion exchanger 1 (AE1), and chloride channel 3 (ClC-3), were double stained with NKA, the basolateral marker of branchial mitochondrion-rich cells (MRCs), to reveal the localization of these transporter proteins in gill MRC of FW- or SW-acclimated milkfish and pufferfish. Confocal microscopic observations showed that the localization of these transport proteins in the gill MRCs of the two studied species were similar. However, the number of gill NKA-immunoreactive (IR) cells in milkfish and pufferfish exhibited to vary with environmental salinities. An increase in the number of NKA-IR cells should lead to the elevation of NKA activity in FW milkfish and SW pufferfish. Taken together, the opposite branchial NKA responses observed in milkfish and pufferfish upon salinity challenge could be attributed to alterations in the number of NKA-IR cells. Furthermore, the localization of these Cl⁻ transporters in gill MRCs of the two studied species was identical. It depicted the two studied euryhaline species possess the similar Cl⁻ transport mechanisms in gills.     

Influence of salinity on the energetics of gill and kidney of Atlantic salmon (Salmo salar)

The effect of seawater acclimation and adaptation to various salinities on the energetics of gill and kidney of Atlantic salmon (Salmo salar) was examined. Smolts and non-smolts previously reared in fresh water were exposed to a rapid increase in salinity to 30 ppt. Plasma osmolarity, [Na⁺], [Cl^–], [K⁺] and [Mg⁺⁺] increased in both groups but were significantly lower in smolts than non-smolts. Gill Na⁺, K⁺-ATPase specific activity, initially higher in smolts, increased in both groups after 18 days in seawater. Kidney Na⁺, K⁺-ATPase specific activity was not affected by salinity in either group. Gill and kidney citrate synthase specific activity was not affected by seawater exposure in smolts but decreased in non-smolts. In a second experiment, Atlantic salmon smolts reared in fresh water were acclimated to 0, 10 or 30 ppt seawater for 3 months at a temperature of 13–14°C. Gill Na⁺, K⁺-ATPase was positively correlated with salinity, displaying 2.5- and 5-fold higher specific activity at 10 and 30 ppt, respectively, than at 0 ppt. Kidney Na⁺, K⁺-ATPase specific activity was not significantly affected by environmental salinity. Citrate synthase and cytochrome c oxidase specific activities in gill were slightly (6–13%) lower at 10 ppt than at 0 and 30 ppt, whereas kidney activities were lowest at 30 ppt. Oxygen consumption of isolated gill filaments was significantly higher when incubated in isosmotic saline and at 30 ppt than at 0 ppt, but was not affected by the prior acclimation salinity. The results indicate that although high salinity induces increased gill Na⁺, K⁺-ATPase activity, it does not induce substantial increases in metabolic capacity of gill or kidney.     

Physiological characteristics and stress resistance of great sturgeon (<Emphasis Type="Italic">Huso huso</Emphasis>) juveniles fed with vitamins C,E, and HUFA-enriched <Emphasis Type="Italic">Artemia urmiana</Emphasis> nauplii

This study was carried out to examine the effect of Artemia urmiana nauplii enriched with HUFA, and vitamins C and E on stress tolerance, hematocrit, and biochemical parameters of great sturgeon, Huso huso juveniles. Cod liver oil (EPA 18% and DHA 12%), ascorbyl-6-palmitate and α-tocopherol acetate were used as lipid, and vitamin C and E sources, respectively. Beluga juveniles at the stage of first feeding (69.7 ± 5.9 mg body weight) were randomly divided into five treatments and three tanks were assigned to each diet. All fish groups were fed non-enriched Artemia for the initial 5 days and then fed enriched Artemia for 7 days. Juveniles were fed with Artemia enriched with HUFA + 20% vitamin C (C group); HUFA + 20% vitamin E-enriched Artemia nauplii (E group); HUFA + 20% vitamin C + 20% vitamin E (C and E group); HUFA without vitamins (HUFA) and non-enriched Artemia (control). After the period of enrichment, Juveniles were fed with Daphnia sp. from the 13th to the 40th day. At day 40, the fish were transferred directly from fresh water (0.5 ppt) to brackish water (6 ppt for 4 days and 12 ppt for 2 days) and warm water (from 27 to 33°C) to evaluate juvenile resistance to salinity and thermal shocks. Moreover, all treatments were separately exposed to freshwater in tanks with the same capacity as used for osmotic and thermal tests (as fresh water control). The addition of vitamins C, E, and C + E to HUFA significantly increased fish resistance to 12 ppt salinity and temperature stress tests, whereas survival was not significantly different among challenges at 6 ppt. There was no significant difference in the hematocrit index under stress conditions. Enrichment had significant influence on plasma Na⁺ level in the C group on the 4th day at 6 ppt. Na⁺ and Ca²⁺ concentrations in C, E, and C and E groups on the 1st day at 12 ppt, and Ca²⁺ level in E group on the 2nd day at 12 ppt were lower than the other groups. The glucose level in the C and C and E groups was lower than the other treatments on the 1st day at 12 ppt and the 2nd day at 33°C. Regardless of Artemia enrichment, plasma ions (Na⁺, K⁺, Ca²⁺, and Mg²⁺) and glucose concentrations in fish exposed to salinity stress tests were higher than fish in fresh water. Glucose concentration in plasma also increased after 2 days at 33°C. Although most of our results were not significantly different, the use of vitamins C, E, and HUFA in Artemia enrichment can improve Juveniles tolerance under stress conditions, and regardless of enrichment, these data show that beluga juveniles are partly sensitive to high salinity and temperature.     

Interactive effects of salinity on oxygen consumption,ammonium excretion,osmoregulation and Na+/K+‐ATPase expression in the bullseye puffer (Sphoeroides annulatus,Jenyns 1842)

A physiological study was performed to explore the effects in Sphoeroides annulatus (Jenyns 1842) juveniles of exposure to different salinities 10, 17, 23, 29, 35 and 41‰ on oxygen consumption rate (OCR), ammonium excretion rate (AER), the oxygen‐nitrogen atomic ratio (O:N), osmoregulation and Na⁺/K⁺‐ATPase expression. The OCR values ranged from 27.9 to 30.9 mg O₂ h^?1 kg^?1, displaying a Type 1 response pattern Kinne (1977). AER ranged from 0.60 to 0.69 mg h^?1 kg^?1, and O:N values were from 53.12 to 59.26, indicating that the puffers use proteins and lipids as an energy‐substrate. Osmoregulation in S. annulatus was hyposmotic in salinities of 23, 29, 35 and 41‰ and hyperosmotic in a salinity of 10‰. The isosmotic point of the bullseye puffer was 356 mmol kg^?1 (10.5‰). The enzyme expression analysis indicates that the fish acclimated to 41‰ had the highest Na⁺/K⁺‐ATPase expression level, whereas, the lowest expression level was found close to the isosmotic point. Na⁺/K⁺‐ATPase expression in the gills was found to have a U‐shaped relationship with environmental salinity. We conclude that the bullseye puffer is strongly euryhaline and can be cultivated in a wide range of environments that has important economic implications.     

Osmoregulation in juvenile Chinese sturgeon (<Emphasis Type="Italic">Acipenser sinensis</Emphasis> Gray) during brackish water adaptation

The osmoregulation capabilities of 7-month-old juvenile Chinese sturgeon (Acipenser sinensis Gray) (128.8 ± 15 g) transferred directly from fresh water (0‰, 46 mOsmol kg⁻¹) to brackish water (10‰, 273 mOsmol kg⁻¹) were studied over a 20-day period. Changes in serum osmolarity, chloride (Cl⁻), sodium (Na⁺), potassium (K⁺) and calcium (Ca²⁺) ion concentrations, as well as gill and spiral valve Na⁺,K⁺-ATPase activities were measured at 3, 12, 24, 72, 216 and 480 h after transfer to BW. The serum osmolarity and ion concentrations (Na⁺, Cl⁻ and Ca²⁺) increased immediately after the transference to BW, reaching maximum at 24 h and returned to a new steady state at 216 h, while the FW control group maintained basal levels which showed lower (P < 0.05) than the BW group. Gill Na⁺,K⁺-ATPase activity of BW group exhibited an abrupt decrease in the first 3 h after transfer, but began to increase at 3 h, reaching a peak value at 24 h, and returned to a new steady state at 216 h. The differences between gill Na⁺,K⁺-ATPase activity of BW and FW fish were significant (P < 0.05) after 12 h. In contrast, Na⁺,K⁺-ATPase activity of the spiral valve showed transient increase after transference from FW to BW, and then decreased rapidly at 3 h, reaching the lowest at 24 h after transference. At 216 h after exposure to BW, Na⁺,K⁺-ATPase activities of the spiral valve increased slowly to the levels of FW control. The results of our study indicate the existence of hyposmoregulatory adaptive mechanisms in 7-month-old juvenile Chinese sturgeon which enable this fish to acclimate itself successfully to brackish water.