Salt uptake and shoot water relations in mangroves

The effects of salinity on water relations and ion concentrations were investigated in seedlings of the mangroves Avicennia alba, Bruguiera gymnorrhiza, Heritiera littoralis and Xylocarpus granatum grown at salinities of 0, 10, 20, 30, 40 and 60‰. All four species survived and grew at salinities ranging from 0 to 40‰, but none survived at a salinity of 60‰. The concentration of sodium and chloride in the xylem sap increased with increasing salinity in both A. alba and B. gymnorrhiza. Sodium and chloride concentrations in the xylem sap of A. alba grown at 40‰ salinity both reached 114 mol m⁻³, about 15% of the external concentration around the roots. The xylem sap of B. gymnorrhiza grown at 40‰ salinity, by contrast, contained only 7.0 mol m⁻³ sodium and 4.1 mol m⁻³ chloride, about 1% of their concentrations in the external solution around the roots. The results indicated that B. gymnorrhiza, which does not have salt-secreting glands, was more effective at excluding salt than A. alba, which has salt-secreting glands.Analysis of pressure–volume curves showed that the bulk modulus of elasticity increased with increasing salinity. This was accompanied by a decrease in shoot water potential, mainly associated with a reduction in shoot osmotic potentials with increasing salinity. The decrease in osmotic potential was attributed to increasing solute concentrations, particularly sodium and chloride, in the leaves of all species except H. littoralis, which had little sodium and chloride in the leaves.     

The rôle of ninhydrin-positive substances in osmoregulatton in the western rock lobster,Panuliris longipes (Milne Edwards)

Free leucine, valine, proline, tyrosine, alanine, taurine, glutamic acid, and glycine are present in both blood and muscle of Panulirus longipes (Milne Edw.). Arginine is present in muscle only. Five unidentified ninhydrin-positive spots appeared in thin-layer chromatograms of blood, and four in chromatograms of muscle.Non-protein, ninhydrin-positive substances (NPS) in muscle do not appear to function in the moulting process. Concentrations averaged 278 mM/kg wet weight of muscle and NPS were the most abundant of organic osmotically active substances (total ions were 325 mM/kg). Trimethylamine oxide and glycine betaine together amounted to a further 121 mM.With lowered salinity, NPS concentrations in muscle were sharply reduced within 24 h, but with increased salinity, concentrations rose slowly over 7 days suggesting that NPS are produced by intracellular processes. When rock lobsters were fully adapted to salinities ranging from 25 to 45‰ concentrations of muscle NPS were linearly related to external salinity. When the salinity was lowered the blood was initially flooded with NPS, concentrations reaching a maximum at 12 h, and returning to normal after 72 h. Gastric fluid concentrations also rose, and evidence indicates that a large part of the NPS lost from muscle is excreted into the external water via the gastric fluid. At a salinity of 30‰, 56 % of the NPS lost from muscle appears in the external water, the remainder being excreted as ammonia; at 27‰, 82 % of NPS was excreted into the water, the remainder as ammonia.It is concluded that a changing external salinity would cause a continual loss of NPS from the body of a decapod crustacean. Although wasteful of nitrogenous compounds, such a process is of high survival value in permitting osmotic adjustment in the absence of more efficient mechanisms, such as those present in teleost fishes.     

Mechanism of salt tolerance in wild rice (Oryza coarctata Roxb)

Summary Oryza coarctata, a highly salt-resistant wild rice species, is commonly found on the banks of coastal rivers in India. This species can also withstand saline water (20 to 40 dSm⁻¹ E.C) submergence for quite a long period. It was revealed thatO. coarctata has some special unicellular salt hairs (trichomes) on the adaxial surface of the leaves, by which they efficiently maintain a low concentration of toxic salts in the plant tissue. Sodium and chloride were the dominant ions in the excreted material but they also excrete potassium, magnesium and calcium. With the increase in soil salinity sodium, magnesium and chloride excretion increased.O. coarctata maintained the optimum mineral concentration in its tissues. Maximum accumulation of potassium was observed in the leaves. With the increase in salt stress total biomass production and osmotic potential increased over control but there was no change in the moisture percentage of leaves.     

Ionic and osmotic regulation of the haemolymph of the dragonfly, Libellula quadrimaculata (Odonata : Libellulidae)

Larvae of the widespread dragonfly, Libellula quadrimaculata, were adapted to a series of salt solutions, and the osmotic pressure, and sodium, potassium and chloride concentrations in the haemolymph measured. The regulation of potassium is extremely efficient over the range 0–50 m-mole/l. external concentration. Above this, larvae die. Sodium and chloride are regulated to a lesser extent, the larvae being able to withstand considerable changes in the concentration of these ions in the haemolymph. However, at higher external concentrations, the haemolymph concentration of these ions is maintained below that of the external medium. The osmotic pressure is regulated in parallel with sodium concentration over most of the range tested. However, in higher salinities, the osmotic pressure of the haemolymph does not fall below that of the external medium. This is seen as a strategy to limit the amount of drinking in saline media. Overall, the osmoregulatory system of L. quadrimaculata resembles that of brackish-water insects, rather than that of the more strictly freshwater dragonflies that have been studied.     

Osmoregulatory strategies in natural populations of the black-chinned tilapia Sarotherodon melanotheron exposed to extreme salinities in West African estuaries

The effect of salinity was studied in natural populations of the black-chinned tilapia (Sarotherodon melanotheron) from West Africa. This euryhaline species colonizes nearly all coastal environments from bays to lagoons characterized by salinities ranging from fresh water to hypersaline water over 100 ‰. Individuals were sampled during the dry season at several locations characterized by different levels of salinity (3-102 ‰). Their osmotic status and their gills were analyzed. The branchial mitochondria-rich cells (MRC), localized at the basis of the filaments and along the lamellae in fish taken from the saline stations, showed a wide plasticity with significant differences in their number and size. The most striking results were a significant larger area (≈3x) and a higher number (≈55x) of MRC at high salinity (102 ‰) compared to low salinity (3 ‰). The major ion transporters and channels were localized by immunocytochemistry and different expression patterns have been recorded between stations. Despite an increased Na?/K?-ATPase (NKA) α-subunit expression and NKA activity, pointing to an increased monovalent ion excretion, a severe osmotic imbalance was recorded in animals living in hypersaline environments.     

Ecophysiology of Salt Excretion in Aeluropus litoralis (Graminae)

Various aspects of salt excretion from leaves of Aeluropus were investigated. Salt excretion exhibited an optimum-type of curve when measured against external salt concentration, while sodium content of the leaves increased linearly. The ‘relative excretion’, i.e. rate of excreted ions: change in leaf ion content, was maximal in the low salt concentration range, and decreased when external sodium chloride concentration increased. Concentration of the excreted droplets was higher than the external concentration when the leaves were exposed to low salt concentrations in the medium, but the reverse occurred when the external salt concentrations were high. The excretion process was sensitive to water-stress conditions, caused either by high external salt concentrations or by exposure to dry atmosphere. A considerable fraction of the leaf sodium content in salt-treated leaves was only slightly available for excretion. Salt excretion in Aeluropus was enhanced by light. Such enhancement was indirect and is attributed to the increase of salt transport via transpiration stream. Selectivity of the salt-excretion mechanism is in favour of sodium and against potassium. On the other hand, potassium has a high affinity for the accumulation systems within the leaves. The ecological significance of the results is discussed.     

Dog Red Blood Cells : Adjustment of salt and water content in vitro

Dog red blood cells (RBC) lack a ouabain-sensitive sodium pump, and yet they are capable of volume regulation in vivo. The present study was designed to find in vitro conditions under which dog RBC could transport sodium outward, against an electrochemical gradient. Cells were first loaded with sodium chloride and water by preincubation in hypertonic saline. They were then incubated at 37°C in media containing physiologic concentrations of sodium, potassium, chloride, bicarbonate, glucose, and calcium. The cells returned to a normal salt and water content in 16–20 h. Without calcium in the medium the cells continued slowly to accumulate sodium. Removal of glucose caused rapid swelling and lysis, whether or not calcium was present. The net efflux of sodium showed a close relationship to medium calcium over a concentration range from 0 to 5 mM. Extrusion of salt and water was also demonstrated in fresh RBC (no hypertonic preincubation) when calcium levels in the media were sufficiently raised. The ion and water movements in these experiments were not influenced by ouabain or by removal of extracellular potassium. Magnesium could not substitute for calcium. It is concluded that dog RBC have an energy-dependent mechanism for extruding sodium chloride which requires external calcium and is quite distinct from the sodium-potassium exchange pump.     

Chronic exposure to soil salinity in terrestrial species: Does plasticity and underlying physiology differ among specialized ground-dwelling spiders?

In salt marshes, the alternation of low and high tides entails rapid shifts of submersion and aerial exposure for terrestrial communities. In these intertidal environments, terrestrial species have to deal with an osmotic loss in body water content and an increase in sodium chloride concentration when salt load increases. In salt marshes, spiders represent an abundant arthropod group, whose physiological ecology in response to variations of soil salinity must be further investigated. In this study, we compared the effect of salinity on the survival and physiology of three species of Lycosidae; two salt marsh species (Arctosa fulvolineata and Pardosa purbeckensis) and one forest species (P. saltans). Spiders were individually exposed at three salinity conditions (0‰, 35‰ and 70‰) and survival, changes in body water content, hemolymph ions (Na⁺, Ca²⁺, Mg²⁺, K⁺; ICP-MS technique) and metabolites (mainly amino acids, polyols, sugars; LC and GC techniques) were assessed. The survival of the forest species P. saltans was very quickly hampered at moderate and high salinities. In this spider, variations of hemolymph ions and metabolites revealed a quick loss of physiological homeostasis and a rapid salt-induced dehydration of the specimens. Conversely, high survival durations were measured in the two salt-marsh spiders, and more particularly in A. fulvolineata. In both P. purbeckensis and A. fulvolineata, the proportion of Na⁺, Ca²⁺, Mg²⁺, K⁺ remained constant at the three experimental conditions. Accumulation of hemolymph Na⁺ and amino acids (mainly glutamine and proline) demonstrated stronger osmoregulatory capacities in these salt-marsh resident spiders. To conclude, even if phylogenetically close (belonging to the same, monophyletic, family), we found different physiological capacities to cope with salt load among the three tested spider species. Nevertheless, physiological responses to salinity were highly consistent with the realized ecological niches of the spiders.     

Intra- and inter-cultivar impacts of salinity stress on leaf photosynthetic performance,carbohydrates and nutrient content of nine indigenous Greek olive cultivars

Nine indigenous Greek olive cultivars (‘Aetonicholia Kynourias’, ‘Arvanitolia Serron’, ‘Ntopia Atsicholou’, ‘Koroneiki’, ‘Lefkolia Serron’, ‘Ntopia Pierias’, ‘Petrolia Serron’, ‘Smertolia’ and ‘Chryssophylli’) were evaluated for their tolerance to salinity stress (four levels of sodium chloride salt, i.e., 0, 50, 100 and 200 mM) under hydroponic conditions. Their photosynthetic performance, leaf carbohydrates (mannitol, glucose, fructose and sucrose) and nutrients (nitrogen, potassium, calcium, sodium and chloride) were assessed. Photosynthetic performance was reduced under salt stress and this was mostly evident in ‘Koroneiki’ and ‘Ntopia Atsicholou’ (approximately 20% of the corresponding controls), while ‘Ntopia Pierias’, ‘Smertolia’ and ‘Petrolia Serron’ did not exhibit significant changes with salinity level. Photosynthesis (A) was reduced mainly due to severe stomatal limitations. A weak correlation was detected between A and intercellular CO₂ (Ci) indicating a minor role of non-stomatal limitations. Carbohydrates in the leaves did not seem to undergo significant changes. Mannitol accumulated in ‘Chryssophylli’ leaves and glucose in ‘Arvanitolia Serron’ leaves under the highest salinity level. Potassium concentration per leaf water volume was significantly reduced (especially under the highest salinity level ?45 to 60% of control). Calcium was not significantly affected although Ca/Na ratio was reduced, due to the great increase of sodium concentration. ‘Lefkolia Serron’ and ‘Arvanitolia Serron’ accumulated the least sodium in their leaves, exhibiting high K/Na ratio under the highest salinity level, indicating a better regulation of potassium influx under high sodium concentration. Based on the present data and on previous research ‘Lefkolia Serron’ and ‘Arvanitolia Serron’ are the two cultivars with the highest tolerance against salinity stress.     

Plasma thyroxine and cortisol profiles and gill and kidney Na+/K+-ATPase and SDH activities during acclimation of the catfish Heteropneustes fossilis (bloch) to higher salinity, with special reference to the effects of exogenous cortisol on hypo-osmoregulatory ability of the catfish

When the stenohaline catfish Heteropneustes fossilis was transferred from fresh water (FW) to 30% seawater (SW), the Na(+)/K(+)-ATPase activity significantly increased in the kidney, while in gills it remained more or less constant. A reverse pattern was observed for succinic dehydrogenase (SDH) activity inasmuch as it significantly increased in gills and remained unchanged in the kidney. Plasma osmolality significantly increased within 3 days of transfer to 30% SW and remained significantly higher throughout the duration of experiment. These results suggest that catfish gills may not be able to reverse their function from salt uptake in FW to salt excretion at higher salinity, and that the elimination of monovalent as well as divalent ions is performed by the kidney but not the gills. The significant decline in plasma cortisol (F) levels following transfer to higher salinity may not be due to reduced production but rather to an enhanced utilization and clearance rate, a conclusion supported by the fact that exogenous administration of cortisol acetate (FA) resulted in significant increases in branchial and renal Na(+)/K(+)-ATPase in FW and 30% SW. FA also improved the plasma osmotic regulatory ability of the catfish, possibly due to a change in branchial function from salt-absorption to salt excretion, as was evident from a significant increase in branchial Na(+)/K(+)-ATPase activity in the fish in 30% SW pretreated with FA for 5 days. Consistently higher levels of plasma thyroxine (T4) following transfer to higher salinity suggest the involvement of this hormone at higher salinity.     

Respiratory and osmoregulatory responses of the hermit crab, Clibanarius vittatus (Bosc), to salinity changes

Intertidal hermit crabs were stepwise acclimated to 10, 20, and 30‰ salinity (S) and 21 ± 1 °C. Hemolymph osmolality, sodium, chloride, and magnesium were isosmotic (isoionic) to ambient sea water at 30‰ and hyperosmotic (hyperionic) at 20 and 10‰ S, while hemolymph potassium was significantly hyperionic in all acclimation salinities. Total body water did not differ significantly at any acclimation salinity. Oxygen uptake rates were higher in summer-than winter-adapted crabs. No salinity effect on oxygen consumption occurred in winter-adapted individuals. Summer-adapted, 30‰ acclimated crabs had a significantly lower oxygen consumption rate than those acclimated 10 and 20‰ S. Crabs exposed to 30 10 30‰ and 10 30 10‰ semidiurnal (12 h) and diurnal (24.8 h) fluctuating salinity regimes showed variable osmoregulatory and respiratory responses. Hemolymph osmolality followed the osmolality of the fluctuating ambient sea water in all cases, but was regulated hyperosmotically. Hemolymph sodium, chloride, and magnesium concentrations were similar to hemolymph osmolality changes. Sodium levels fluctuated the least. Hemolymph potassium was regulated hyperionically during all fluctuation patters, but corresponded to sea water potassium only under diurnal conditions. The osmoregulatory ability of Clibanarius vittatus (Bosc) resembles that reported for several euryhaline brachyuran species. The time course of normalized oxygen consumption rate changed inversely with salinity under semidiurnal and diurnal 10 30 10‰ S fluctuations. Patterns of 30 10 30‰ S cycles had no effect on oxygen consumption rate time course changes. The average hourly oxygen consumption rates during both semidiurnal fluctuations were significantly lower than respective control rates, but no statistical difference was observed under diurnal conditions.     

Teleost chloride cell. I. Response of pupfish Cyprinodon variegatus gill Na,K-ATPase and chloride cell fine structure to various high salinity environments

Certain euryhaline teleosts can tolerate media of very high salinity, i.e. greater than that of seawater itself. The osmotic gradient across the integument of these fish is very high and the key to their survival appears to be the enhanced ability of the gill to excrete excess NaCl. These fish provide an opportunity to study morphological and biochemical aspects of transepithelial salt secretion under conditions of vastly different transport rates. Since the cellular site of gill salt excretion is believed to be the "chloride cell" of the branchial epithelium and since the enzyme Na,K-ATPase has been implicated in salt transport in this and other secretory tissues, we have focused our attention on the differences in chloride cell structure and gill ATPase activity in the variegated pupfish Cyprinodon variegatus adapted to half-strength seawater (50% SW), seawater (100% SW), or double-stregth seawater (200% SW). The Na,K-ATPase activity in gill homogenates was 1.6 times greater in 100% SW. When 50% SW gills were compared to 100% SW gills, differences in chloride cell morphology were minimal. However, chloride cells from 200% SW displayed a marked hypertrophy and a striking increase in basal-lateral cell surface area. These results suggest that there are correlations among higher levels of osmotic stress, basal-lateral extensions of the cell surface, and the activity of the enzyme Na,K-ATPase.     

Plasma homeostasis and cloacal urine composition inCrocodylus porosus caught along a salinity gradient

Summary Juveniles of the Estuarine or Saltwater Crocodile,Crocodylus porosus, maintain both osmotic pressure and plasma electrolyte homeostasis along a salinity gradient from fresh water to the sea. In fresh water (FW) the cloacal urine is a clear solution rich in ammonium and bicarbonate and containing small amounts of white precipitated solids with high concentrations of calcium and magnesium. In salt water (SW) the cloacal urine has a much higher proportion of solids, cream rather than white in colour, which are the major route for excretion of potassium in addition to calcium and magnesium. Neither liquid nor solid fractions of the cloacal urine represent a major route for excretion of sodium chloride. The solids are urates and uric acid, and their production probably constitutes an important strategy for water conservation byC. porosus in SW. These data, coupled with natural history observations and the recent identification of lingual salt glands, contribute to the conclusion thatC. porosus is able to live and breed in either fresh or salt water and may be as euryhaline as any reptile.     

Effect of elevated temperature on osmotic and ionic regulation in a salt-tolerant caddisfly from Death Valley,California

The effects of temperature (8–10 or 20°C) on regulation of haemolymph osmotic and ionic concentrations were investigated over a range of salinities (0–25‰) in fifth-instar larvae of the Death Valley caddisfly Limnephilus assimilis. At low temperatures, levels of chloride and sodium in the haemolymph are regulated over a wide range of salinities corresponding to the salinities at which larvae occur in nature and at which they can complete development into adults. In contrast, haemolymph osmolality is constant at low salinities (<14‰) but approaches conformity with the medium at higher salinities. High temperature reduces the larva's ability to maintain low chloride concentrations in its haemolymph and also leads to a reduction in haemolymph osmotic pressure; thus, at high temperatures ions account for more of the haemolymph osmotic concentration than at low temperatures. These data suggest that the absence of larvae from thermal pools and from all Death Valley waters in summer can be explained by the effects of high water temperatures on hydromineral regulation.     

Response of Bean Plants to Sodium Chloride and Sodium Sulphate Salinization

Bean plants were grown under constant levels of sodium chlorideand sodium sulphate salinity, and under changing levels of sodiumsulphate salinity. Although growth was suppressed similarly by the two types ofsalinity when expressed on an osmotic basis, other parametersshowed different responses according to salinity type. Chloride-salinatedplants have thicker leaves with higher water content than thesulphate and control plants. The relative water content of thesulphate plants was somewhat lower. Transpiration rates weresuppressed more by chloride salinity. Osmotic adjustment seemsto be faster under chloride salinity and was of a differentnature. Chloride accumulated to much higher levels than sulphate.Increase in potassium ion concentration and decrease in calciumion concentration was more pronounced under sulphate salinity,but the total cation concentration in the sap was similar forall treatments. As a result, the inorganic ions' electricalbalance was more negative under chloride salinity. Leaf expansion in the changing-level treatment reflected rapidlythe variation m substrate salinity. However, total yield wassuppressed by the changing treatments to a similar extent asby constant salinity. According to other parameters the changingregime may be considered as a shorter period of exposure tosalinity.     

The osmotic behaviour of frogs eggs and young tadpoles

Summary Determinations of total osmotic concentration on eggs ofRana temporaria by the vapour pressure method and of chloride by theWigglesworth ultra micro-technique show a rapid fall during the first few hours, from 120 to 95 mM total concentration and, until the blastopore closes, a further slight fall to about 80 mM. Thereafter the total concentration rises rapidly, while the chloride concentration remains unaltered or (later) rises very gradually.The eggs swell considerably at first and then more gradually and the changes in concentration until the budding of the gills are due at least in the main to osmotic uptake of water.The permeability, as determined both by rate of swelling and in special experiments with heavy waterbecomes greatly reduced by fertilization.The initial permeability is calculated to correspond to a minute Number of 140 days, while later it is reduced to about 5 1/2 years.When the gills begin to develop permeability for water again increases and the weight rises by osmotic inflow of water. The kidneys become functional and an active uptake of salt (probably located in the gills) prevents a reduction of the osmotic concentration.The fluid in the chorionic cavity is very slightly hypertonic to the surrounding water.The degree of swelling of the egg mucus depends upon the salt concentration in the surrounding water. The swelling is enormous in distilled water and seems to be specifically inhibited by calcium.     

The role of the antennal glands in ion and body volume regulation of cannulated Penaeus monodon reared in various salinity conditions

Urinary production rate and the osmotic and ionic concentrations in both urine and hemolymph were measured in cannulated intermolt Penaeus monodon which were either abruptly transferred from 45 ppt seawater to 15 ppt seawater (Experiment 1) or acclimated to 5, 25 and 45 ppt seawater (Experiment 2). In Experiment 1, urinary magnesium concentration fell dramatically from 228 to 30 mEq/l within 4 h post-transfer, but 8 h after transfer, U/H (urine/hemolymph) ratios stabilized at between 1.0 and 2.5. Sodium was higher in urine than in hemolymph during the first 24 h after transfer, while potassium was lower in urine than in hemolymph until 72 h after transfer, which suggests that sodium and potassium concentrations are regulated by the antennal gland after an abrupt change in media. In Experiment 2, the urinary production rate of P. monodon decreased as salinity increased, suggesting that the antennal glands also regulate body volume. In the acclimated shrimps of Experiment 2, the antennal glands did not appear to regulate osmolarity or the concentration of chloride, sodium, potassium, and calcium ions, but as salinity increased, U/H ratios of magnesium increased from 2.3 to 13.5, and active secretion by the antennal gland accounted for 57 approximately 93% of the total magnesium excretion through urine. These results suggest that active secretion of magnesium by the antennal gland enable this shrimp to maintain hypoionic levels of magnesium in the hemolymph.     

Hydromineral regulation in the saline water corixid Trichocorixa reticulata (Hemiptera: Corixidae)

The aquatic corixid Trichocorixa reticulata (Guerin-Meneville) inhabits coastal marshes, brackish water ponds and salt ponds of high salinity, suggesting the presence of well developed mechanisms for hydromineral regulation.Groups of corixids acclimated in salinities ranging from fresh water to just above 300% sea water (100‰) were analyzed for total body water content, haemolymph ionic and osmotic levels, and haemolymph free amino acids.Results indicate an excellent ability to maintain haemolymph Na⁺, Cl^?, Mg²⁺ and K⁺ hyperosmotic to the medium at low salinities and hyposmotic at high salinities. Calcium appears to conform closely to changes in external medium, becoming hyposmotic at very high salinities (80‰).Total haemolymph osmotic pressure was well regulated, the freezing point depression varying from 0.75°C in distilled water to 1.15°C in salinities of 100‰. Total body water was maintained at approx. 75% of the total animal wet weight at all salinities tested.Free amino acids were maintained between 40–60 mM in all tests and did not appear to change with salinity.     

Metallothionein-like proteins in the blue crab Callinectes sapidus: Effect of water salinity and ions

The effect of water salinity and ions on metallothionein-like proteins (MTLP) concentration was evaluated in the blue crab Callinectes sapidus. MTLP concentration was measured in tissues (hepatopancreas and gills) of crabs acclimated to salinity 30 ppt and abruptly subjected to a hypo-osmotic shock (salinity 2 ppt). It was also measured in isolated gills (anterior and posterior) of crabs acclimated to salinity 30 ppt. Gills were perfused with and incubated in an isosmotic saline solution (ISS) or perfused with ISS and incubated in a hypo-osmotic saline solution (HSS). The effect of each single water ion on gill MTLP concentration was also analyzed in isolated and perfused gills through experiments of ion substitution in the incubation medium. In vivo, MTLP concentration was higher in hepatopancreas than in gills, being not affected by the hypo-osmotic shock. However, MTLP concentration in posterior and anterior gills significantly increased after 2 and 24 h of hypo-osmotic shock, respectively. In vitro, it was also increased when anterior and posterior gills were perfused with ISS and incubated in HSS. In isolated and perfused posterior gills, MTLP concentration was inversely correlated with the calcium concentration in the ISS used to incubate gills. Together, these findings indicate that an increased gill MTLP concentration in low salinity is an adaptive response of the blue crab C. sapidus to the hypo-osmotic stress. This response is mediated, at least in part, by the calcium concentration in the gill bath medium. The data also suggest that the trigger for this increase is purely branchial and not systemic.