Salinity effects on oxygen consumption, gill Na+, K+-ATPase and ion regulation in juvenile coho salmon

The metabolic response of juvenile coho salmon Oncorhynchus kisutch to different salinities was examined, using whole-animal oxygen consumption rates and gill Na⁺, K⁺-ATPase activities as indicators of osmoregulatory energetics. Coho salmon smolts were acclimated to fresh water (FW), isosmotic salinity (ISO, 10‰) and sea water (SW, 28‰) and were sampled for up to 6 weeks for plasma levels of cortisol, glucose and ions (Na⁺, K⁺, Cl⁻), gill Na⁺, K⁺-ATPase activity and oxygen consumption rates. Following an initial adjustment period, plasma constituents in SW fish returned to near-FW values, indicating that the fish were acclimated to SW by day 21. Gill Na⁺, K⁺-ATPase activities on days 21 and 42 were lowest in ISO, higher in FW and highest in SW. This result is consistent with the idea that less energy would be required to maintain ion balance in an isosmotic environment, where the ionic gradients between extracellular fluid and water would be minimal. Oxygen consumption rates of swimming fish (1 body length s⁻¹), however, did not differ significantly between the three test salinities after 6 weeks. The results of this study suggest that the metabolic response of juvenile salmonids to changes in salinity is dependent on life-history stage (e.g. fry v . smolt), and that oxygen consumption rates do not necessarily reflect osmoregulatory costs.     

Ionic regulation and Na+,K+-ATPase activity in gills and kidney of the Antarctic aglomerular cod icefish exposed to dilute sea water

When Notothenia neglecta was exposed to diluted, half strength, sea water for 6 h or 10 days, serum concentrations of Cl^-, Na⁺, K⁺ and Mg²⁺ did not differ from those of sea water controls. This indicates that the fish were capable of both short- and long-term regulation. Renal Na⁺,K⁺-ATPase activity decreased after a 6 h exposure to diluted sea water, but there were no differences between diluted sea water and controls after 10 days of exposure.     

Salinity adaptation in the snakehead, Ophiocephalus maculatus Lacépède: changes in oxygen consumption, branchial Na+-K+-ATPase and body composition

Snakeheads were adapted to fresh water (1 mOsm kg⁻¹), 25% sea water (230 mOsm kg⁻¹), 33% sea water (320 mOsm kg⁻¹) and 40% sea water (380 mOsm kg⁻¹) for 20 days. Exposure to salt water resulted in tissue dehydration, elevations of plasma osmolality, Na⁺, Mg²⁺, Cl⁻and protein concentrations and stimulation of branchial Na⁺-K⁺-ATPase activity. These changes were accompanied by concomitant decline of the hepatosomatic index and liver glycogen concentration. The routine rate of oxygen consumption was increased in snake-heads adapted to 33% sea water. These data were taken to indicate a stressful effect of salinity to the snakehead despite documentation of its ability to penetrate into brackish waters.     

Plasma ionic regulation and gill Na+/K+-ATPase changes during rapid transfer to sea water of yearling rainbow trout, Salmo gairdneri: time course and seasonal variation

Danish rainbow trout, Salmo gairdneri Richardson, (40–65 g) were transferred to 28%o sea water at intervals throughout the early spring and summer. Gill Na⁺/K⁺-ATPase of fish kept in fresh water surged distinctly during May. Simultaneously, a body silvering occurred and plasma concentrations of Cl⁻, Na⁺ and total thyroxine (T4) decreased. The seawater transfer-induced adaptive response in plasma electrolytes comprised a biphasic change, i.e., an adjustive peak phase and a regulatory phase lasting for 2 days and 1 week after transfer, respectively. Further, gill Na⁺/K⁺-ATPase activity increased to a new level after an initial lag phase of 2–3 days, but electrolyte regulation was mostly initiated prior to the adaptive change in ATPase activity. In spite of increasing pre-transfer freshwater Na⁺/K⁺-ATPase activity during the spring, the electrolyte peak level, the degree of muscle dehydration and the mortality of fish transferred to sea water increased from April to July. The apparent uncoupling of freshwater Na⁺/K⁺-ATPase activity and plasma electrolyte regulation in sea water is discussed in relation to smelting and prediction of hypo-osmoregulatory performance.     

Effect of Fish Oil Diet on Fatty Acid Composition of Phospholipids of Brain Membranes and on Kinetic Properties of Na+, K+-ATPase Isoenzymes of Weaned and Adult Rats

Abstract: The influence of dietary (n-3) fatty acids (such as eicosapentaenoic and docosahexaenoic acids) as found in fish oil on Na⁺ sensitivity and ouabain affinity of Na⁺, K⁺-ATPase isoenzymes (α1, α2, α3) was studied in whole brain membranes from weaned and adult rats fed diets for two generations. The long chain (n-3) fatty acids supplied by fish oil decreased the fatty acids of the (n-6) series compared with the standard diet, resulting in a decrease in the (n-6)/(n-3) molar ratio in both 21 - and 60-day- old rats. On the basis of ouabain titration, three inhibitory processes with markedly different affinities were associated with isoenzymes, i.e., low affinity (α1), high affinity (α2), and very high affinity (α3). It appears that the fish oil diet, in part via the modification of membrane fatty acid composition, altered the proportion and ouabain affinity of isoenzymes. Na⁺ sensitivity is the best criterion of physiologic change induced by fish oil diet. We calculated the Na⁺ activation for each isoenzyme and found one Na⁺ sensitivity and two Na⁺ sensitivities per isoenzyme in weanling and adult rats fed different diets, respectively. In contrast to α2 and α3, α1 appears insensitive to membrane change induced by fish oil diet. Fish oil diet, which is known to confer cardioprotection, induced significant modulation of Na⁺, K⁺-ATPase isoenzymes at the brain level.     

Development of Sea Urchin Embryos in Artificial Sea Water Containing Br−in Place of Cl−

In artificial sea water in which the Cl⁻concentration was reduced to less than 10% of that in normal sea water by its replacement with Br⁻, sea urchin eggs were fertilized and developed into abnormal plutei following almost the same time schedule as in natural sea water. These embryos had poorly developed spicules, short pluteus arms, somewhat jagged embryo-walls and quasi-normal archenterons. Similar embryos were obtained in another artificial sea water in which 90% of the Cl⁻concentration in normal sea water was reduced by Br⁻and 10% by acetate. In artificial sea water, in which either 90% of the Cl⁻was replaced by Br⁻or 10% was replaced by acetate, embryos developed into plutei with quasi-normal spicules, pluteus arms and archenterons. These findings indicate that deficiency of Cl⁻results in somewhat abnormal sea urchin embryos. When cells derived from isolated micromeres, were cultured in these Cl⁻-deficient artificial sea waters, containing Br⁻in place of more than 70% of the normal Cl⁻concentration in sea water, spicule formation was strongly inhibited, but pseudopodial cables were well developed. Thus, external Cl⁻seems to be necessary for at least normal formation of spicule rods.     

Control of Na+ and K+ transport in Spergularia marina I. Transpiration effects

In this paper we begin our study of factors controlling Na⁺ and K⁺ uptake in the halophyte Spergularia marina (L.) Griseb., with emphasis on plants growing at moderate salinity (0.2x sea water). The involvement of transpiration was considered first because of its potential to account for much or all of the transport of ions, and particularly of Na⁺, to the shoot under these growth conditions. Transpiration was constant with time through most of the light period, quickly dropping to 6% of the day time rate at night. ²²Na⁺ uptake, on the other hand, showed much less day/night variation, and relative transport to the shoot was constant. After establishing that transpiration was linearly related to leaf weight, possible transpiration effects were further considered as correlations between leaf weight and transport to the shoot. Under constant, day-time conditions, with linear effects of time and plant size removed, total transport of ²²Na⁺ to the shoot (per plant) was not correlated to leaf weight. A similar result was found when transport was expressed per gram of root, and when partitioning of total label to the shoot was considered. Finally, the correlation was considered between leaf weight and a Na⁺/K⁺ enrichment factor defined as the Na⁺/K⁺ ratio in the leaves divided by that in the roots. This correlation was also insignificant. The results indicate that analysis of control of Na⁺ and K⁺ uptake and transport in this experimental system need not consider effects of transpiration.     

Relationships between Na+, K+-ATPase, sialic acid content and fluidity in rainbow trout density-separated erythrocytes

Smolt of anadromous Arctic charr Sahelinus alpinus (L.) migrating from Storvatn, northern Norway, had a similar feeding intensity as resident Arctic charr from the benthic- and pelagic zone in early summer in fresh water. While smolts fed mostly on surface insects, however, resident Arctic charr had a more broader diet consisting mostly of chironomids, Bosmina sp., benthic prey and surface insects. This indicates that Arctic charr smolts have adopted a surface oriented feeding behaviour prior to their sea migration.     

Fluxes of Na+, K+ and Cl-, and osmotic adjustment in Lycopersicon pimpinellifolium and L. esculentum during short- and long-term exposures to NaCl

NaCl (140 m M ) was applied to 14-day-old plants of salt-sensitive Lycopersicon esculentum Mill. cv. Volgogradskij and its wild relative L. pimpinellifolium Mill. accession PE-2. Changes in the relative growth rate of whole plant, and in the levels of inorganic and organic solutes in leaves, stems and roots were followed for 15 days after the application. Short-term salt exposure (4–6 days of salinization) resulted in enhanced relative growth rates for L. pimpinellifolium , but did not affect growth of L. esculentum , After 6 days of salinization, the relative growth rates of both species decreased significantly; leading to practically comparable growth rates for them by day 15. In all parts of both species, the contribution of organic solutes to the osmotic potential (Ψ_s) gradually decreased from 30% on day 0 to a value lower than 5% on day 4. In L. pimpinellifolium , compared to L. esculentum , short-term salt exposure resulted in (1) a higher percentage of adjustment of Ψ_s; and (2) increases in Na⁺ and K⁺ uptake rates, and in the levels of organic acids and proline (the level of which reached that of sugars, i.e., 10 μmol g^-1 dry weight. Conversely, in L. esculentum , drastic reductions of K⁺ uptake rates and organic acid levels occurred already on day 1. During long-term salt exposure, both species were able to adjust osmotically and both exhibited decreases in organic acid levels as well as in K⁺ uptake and accumulation rates in all parts. The results are discussed in an attempt to explain the adaptive responses during short-term salt exposure and the metabolic dysfunctions that lead to growth decrease after long-term exposure to salt.     

Interactions between growth, uptake of Cl− and Na+, and water relations of plants in saline environments. II. Highly vacuolated cells

Abstract. In a highly saline environment high rates of ion uptake are required to generate sufficient osmotic pressure to maintain the turgor that is needed for the continued growth of plants. We estimate the rates of net uptake of Cl⁻ and Na⁺ required by growing cells to sustain cell expansion at an external NaCl concentration of 500 mol m⁻³. We also estimate the ion fluxes required to regulate turgor of expanding and fully expanded cells during diurnal changes in transpiration. Passive fluxes could contribute significantly to osmotic regulation, but active fluxes are still essential and would consume a substantial amount of energy. We discuss whether a limitation to growth at high salinity would arise from lack of energy, or from insufficient capacity for ion uptake. There is insufficient evidence to choose between these possibilities.     

The influence of salinity on oxygen consumption and plasma electrolytes in grass carp, Ctenopharyngodon idella Val.

Fingerling grass carp, Ctenopharyngodon idella , 90–160 mm t.l. , were acclimated to experimental salinities of < 1 (fresh water), 91, 195 and 317 mOsm/kg. Oxygen consumption rates of fish declined, as salinity increased, from 0.16 mg O₂/g-h in fresh water to 0.11 mg O₂/g-h at an ambient concentration of 317 mOsm/kg. Plasma electrolytes (Na⁺ and Cl⁻¹) and total plasma ionic concentrations increased slightly following 10 days of exposure to ambient salinities greater than 195 mOsm/kg. At 317 mOsm/kg fish appeared to lose control of plasma electrolyte concentrations. As the osmotic gradient was reduced between the fish and the external medium it might have become advantageous to the maintenance of the osmotic equilibrium to reduce blood circulation to the gills, thus imposing a reduction in metabolic rate.     

Plasma levels of C18-, C19- and C21-steroids in captive and feral female sea bass

Morphometric analysis of the gonads of sea bass Dicentrarchus labrax revealed that captive fish matured 1 month later than feral fish, but levels of gonadal steroids were identical in both groups at the same stage of sexual development. 17β-oestradiol (E₂) (up to 3 ng ml-1) and testosterone (T) (up to 4 ng ml-1) were highest during the gametogenetic period while 17,20β,21-trihydroxy-4-pregnen-3-one (17,20β,21-P) (free and sulphated) were maximal during the spawning period. Free 17,20β-dihydroxy-4-pregnen-3-one (17,20β-P) was very low and did not change (c. 0·5 ng ml⁻¹) while 17,20β-P-sulphate increased during the spawning period in both groups (up to 2 ng ml⁻¹). In contrast cortisol levels were higher in captive fish and increased during the spawning period (up to 100 ng ml⁻¹). These results suggest that captivity delays vitellogenesis and spawning in sea bass without affecting the final levels of the gonadal steroids and further indicates a role for cortisol in the latter period. The increased levels during the spawning period suggests a pheromonal role for 17,20β-P-sulphate and 17,20β,21-P-conjugates and the involvement of 17,20β,21-P in final ooccyte maturation.     

Smolting and seasonal variation in the smolt characteristics of one- and two-year-old Saimaa landlocked salmon under fish farm conditions

Silvering of the skin, reduced condition factor, elevated gill Na⁺, K⁺-ATPase activity and well-developed capacity to regulate the osmotic and ionic balance in sea water were observed in 1 and 2 year old hatchery-reared Saimaa landlocked salmon Salmo salar m. sebago during April-June. Loss of hypoosmoregulatory ability and gill Na⁺K⁺-ATPase activity was observed earlier in 2 year than 1 year old fish. Coincident with changes associated with smolting both age groups showed diminished osmoregulatory capacity in fresh water. Slow growth during May-June may also be attributed to osmoregulatory difficulties in fresh water. These results support the suggestion that the developmental changes at smolting are seasonal and unrelated to any salinity changes and the idea of smolting as evidence of maladaptation of the fish to fresh water.     

The disruption of sodium balance in brook charr, Salvelinus fontinalis (Mitchill), by manganese and iron

A primary toxic action of manganese to brook charr, Sulvelinusjonfinalis, at concentrations near or above the 96 h LC₅₀ was the disruption of sodium regulation. Body and plasma sodium concentrations of brook charr declined by 52 and 40%, respectively, during exposure to 10.9 mM manganese (in 250 PM CaCI), and all fish died within 36 h. Sodium balance was less severely affected by 2.7 and 5.5 mM manganese. An increase in the external calcium concentration from 0.05 to 1.0 mM raised the LC₅₀ for manganese from 4.9 to 5.8 mM, and a further increase to 2.5 mM calcium almost doubled it to 10.2 mM. An examination of stable manganese uptake by the gills revealed that accumulation was inversely correlated with body sodium concentration (r =−0.77). Radioactive J4Mn entered the bloodstream in low levels and accumulated in the liver. Thus manganese may have systemic effects as well as those attributable to surface binding on the gill. Studies of the mechanism ofdissolved iron toxicity were less conclusive, but it did not appear to involve extensive disruption of sodium balance. There was about a 15% drop in body sodium concentration when the trout were exposed for 48 h to the 96 h LC₅₀ level of iron, but plasma sodium was unaffected. Also, an iron concentration at twice the LC₅₀ did not escalate the loss of body sodium, and increasing the water calcium concentration did not raise the LC₅₀.     

Stage Specific Effects on Sea Urchin Embryogenesis of Zn2+, Li+, several Inhibitors of cAMP-Phosphodiesterase and Inhibitors of Protein Synthesis

Treatment of sea urchin embryos for 3 hr starting at the 16-64 cell stage with Li⁺ or 3-isobutyl-1-methylxanthine as well as with other inhibitors of cAMP-phosphodiesterase (PDE) and several inhibitors of protein synthesis, resulted in production of vegetalized embryos with a large exogut. However, the same treatment starting at other stages produced hardly any vegetalized embryos. The specific stage for these substances to cause vegetalization is probably the 16-64 cell stage. Treatment with Zn²⁺ between the times of fertilization and hatching, followed by culture in normal sea water produced animalized embryos with little if any archenteron, but the same treatment followed by culture with ethylenediamine-N, N'-diacetic acid (EDDA), a chelator of Zn²⁺, produced quasi-normal plutei. This chelator did not counteract the animalizing effect of Zn²⁺ when culture with EDDA was started at the post-gastrula stage. Treatment of embryos for a long period (1-3 days) starting at the blastula stage with Li⁺ and the inhibitors of PDE and protein synthesis, as well as with Zn²⁺, produced spherical embryos with little or no archenteron. The stages at which these substances produced abnormal embryos with a poor archenteron are post-hatching stages.     

Response of Suaeda aegyptiaca to KCl, NaCl and Na2SO4 treatments

The response of Suaeda aegyptiaca (Hasselq.) Zoh. to various salinity treatments was tested in sand culture. Growth was promoted by NaCl and by Na₂SO₄ at all tested concentrations, but not by KCl. The effect of NaCl on growth was stronger than that of Na₂SO₄ and it increased gradually up to a 125 eq. m⁻³ optimum. Ion uptake was also affected by the different salts. Cl⁻ was taken up in similar quantities from KCl and from NaCl solutions and the content of the respective cations was also similar to one another. The presence of Na⁺ in the medium lowered the content of K⁺ in the plants and at the same time increased growth by as much as 900%. Transpiration was reduced and water use efficiency increased by Na⁺-salts. Highest water use efficiency was exhibited by plants which were treated with 125 eq. m⁻³ NaCl. It is concluded that Na⁺ at the macronutrient level has a specific promotive effect on the physiological processes of S. aegyptiaca. This effect is not due to replacement of K⁺ by Na⁺; neither can it be achieved by increasing the K⁺ concentration. Cl⁻ has an additional positive effect on growth of S. aegyptiaca. This effect is only expressed in the presence of Na⁺.     

Seasonal changes in the hypo-osmoregulatory ability of brook charr: the role of environmental factors

In winter, brook charr Salvelinus fontinalis demonstrated poor osmoregulatory ability. Saltwater (SW) transfer resulted in sharp increases in plasma osmolality and chloride levels, a decrease in white muscle water content and high mortality. A clear improvement of the hypo-osmoregulatory mechanism e.ciency was observed in spring; by May or June, fish achieved full acclimation within 1 week of SW transfer. In early fall, the time needed to restore osmotic and ionic balance increased once again. A high level of stress was detected only in winter and fall, as indicated by plasma cortisol and glucose concentrations. Great improvement in osmotic and ionic regulation was observed when brook charr were transferred to 80 C SW in February and April while impaired SW adaptability was recorded in individuals transferred to 2·50° C SW in June and August. Temperature itself was more important than a thermal gradient between fresh and salt water. Stress indicators substantiate this temperature-dependent improvement of osmotic and ionic regulation in brook charr. The results support the hypothesis of a minor contribution of cortisol to hypo-osmotic regulation in brook charr, as the cortisol response always seemed to be associated with stress in the experiments.     

Oxygen consumption rates of tilapia in fresh water, sea water, and hypersaline sea water

Whole animal oxygen consumption rates and plasma constituents were determined in the tilapia O. mossambicus , acclimated for 1 month in fresh water, sea water, and 1·6 × sea water. Oxygen consumption rates for the three water salinities were: 177·2 ± 16·86, 78·6 ± 2·32, and 195·4 ± 15·39 mg O₂ kg⁻¹ h⁻¹ (means ± 1 s.e.), respectively. Plasma prolactin (tPRL₁₈₈) concentration was significantly lower in 1·6 × sea water compared to fresh and sea water. There were no significant differences among mean plasma cortisol concentration and lysozyme activity. Ventilation was significantly higher in fish in sea water compared to the fish in fresh and 1·6 × sea water. The lowest oxygen consumption rates were found in fish acclimated to sea water. That salinity is probably closest to the brackish waters from which they were captured in the wild, and this agreement likely reflects the selection for optimal morphological and physiological characteristics to live in that environment.     

Effects of salt stress on H+- ATPase and H+-PPase activities of tonoplast-enriched vesicles isolated from sunflower roots

The control of ion concentration in the cytosol and the accumulation of ions in vacuoles are thought to be key factors in salt tolerance. These processes depend on the establishment in vacuolar membranes of an electrochemical H⁺ gradient generated by two distinct H⁺-translocating enzymes: a H⁺-PPase and a H⁺-ATPase. H⁺-lrans locating activities were characterized in tonoplast-enriched membrane fractions isolated by sucrose gradient centrifugation from sunflower ( Helianthus annuus L.) roots exposed for 3 days to different NaCl regimes. The 15/32% sucrose interface was enriched in membrane vesicles possessing a vacuolar-type H⁺-ATPase and a H⁺-PPase, as indicated by inhibitor sensitivity, pH optimum, substrate specificity, ion effects kinetic data and immunolabelling with specific antibodies. Mild and severe stress did not alter the pH profile, ion dependence, apparent K_m nor the amount of antigenic protein of either enzyme. Saline treatments slightly increased K⁺-stimulaied PPase activity with no change in ATPase activity, while both PP_i-dependent and NO₃-sensitive ATP-dependent H⁺ transport activities were strongly stimulated. These results are discussed in terms of an adaptative mechanism of the moderately tolerant sunflower plants to salt stress.