Tolerance and ultrastructural responses of branchial chloride cells to salinity changes in the euryhaline teleost Oreochromis mossambicus

The changes of intercellular organization and junctional structures in branchial chloride cells reflect respective functions in different salinities. Under TEM, leaky junctions and intercellular digitations occurred between branchial chloride cells of Oreochromis mossambicus Peters adapted to seawater, but not in those adapted to freshwater. The fish transferred directly to 30 S seawater from freshwater died within 6 h, and their chloride cells developed neither leaky junctions nor interdigitations. The fishes acclimated to 20 S seawater for 12 h did not develop the characteristics of seawater-adapted chloride cells and died after transfer to 30 S seawater. The fish acclimated to 20 S seawater for 24 h started to develop seawater-adapted chloride cells, and were able to survive when transferred to 30 S seawater. Thus, the development of leaky junctions and interdigitations in branchial chloride cells appears to correlate to seawater adaptation in O. mossambicus. These changes of seawater-adapted chloride cells seem to be associated with the increase of ion permeability in the gill of teleosts adapted to seawater rather than those adapted to freshwater.     

Branchial Na+−K+-ATPase activity during osmotic adjustments in two freshwater euryhaline teleosts,tilapia (Sarotherodon mossambicus) and orange chromid (Etroplus maculatus)

Effects of osmotic stress on the branchial Na⁺–K⁺-ATPase activity in two freshwater euryhaline teleosts (the tilapia Sarotherodon mossambicus Peters and the orange chromid Etroplus maculatus Bleeker) were studied. Direct transfer from fresh water to salt water with a salinity of 35 S caused extensive mortality in tilapia. Of the remaining four saltwater concentrations (4.375, 8.75, 17.5, and 26.25 S), the higher two increased the enzyme activity significantly in the first week without affecting it further during the remaining seven weeks of acclimation. The lower two concentrations failed to elicit any significant change. In the case of orange chromid, all three salt-water concentrations (4.375, 8.75, and 17.5 S) which the fish survived produced a more significant and extensive change in the enzyme activity, which continued to rise during the entire acclimation period. Employing a procedure of gradual transfer, tilapia and orange chromid were successfully adapted to saltwater concentrations of up to 61.25 and 35 S, respectively. In this experiment, almost each successive increase in salinity elevated the enzyme activity further, but the total change in both species was considerably less extensive than might be expected from the direct transfer study. When the freshwater fish were transferred to identical concentrations of pure NaCl or whole ocean salt, the former produced a relatively greater change in the enzyme activity in tilapia. No such difference was seen in orange chromid. During reacclimation of the saltwater-adapted fish to fresh water, the enzyme activity began to decrease immediately in both species, but remained above the freshwater control levels at the end of eight weeks.     

Prey selection by the hogchoker, Trinectes maculatus (Pisces: Soleidae), along summer salinity gradients in Chesapeake Bay, USA

We investigated feeding by the hogchoker, Trinectes maculatus (Bloch and Schneider), in freshwater, oligohaline, mesohaline, and polyhaline regions of Chesapeake Bay, USA, and examined prey selection in relation to food availability. Otter trawling for fish and Van Veen grab sampling for benthic macrofauna occurred in July and August 1992 and August and September 1993. Hogchokers exhibited both opportunistic and selective feeding patterns along the estuarine salinity gradient in four tributaries (Potomac, Rappahannock, York, and James Rivers) and in the mainstem Chesapeake Bay. Major prey taxa included annelids, arthropods, and tellinid siphons. In polyhaline habitat, polychaetes dominated both the benthos and gut contents numerically and gravimetrically. On the other hand, oligochaetes were numerically dominant in freshwater/oligohaline areas but were rarely eaten, perhaps because of their burial depth. Arthropods (mostly amphipods) occurred at most salinities, were common in gut contents in low-salinity areas, and were replaced as prey by larger proportions of polychaetes in polyhaline regimes. Although hogchokers ate tellinid siphons, they rarely consumed whole bivalves or gastropods. These diet patterns (and especially the importance of siphon nipping) are similar to those of juvenile or small flatfish elsewhere in Europe, Africa, and North America. A size–salinity relationship for hogchokers occurred along the summer salinity gradient, with smaller fish predominating upstream and larger fish downstream. It was not clear from our data if variation in diet composition reflected changes in prey composition along the salinity gradient rather than changes in fish size. Received: 14 June 1997 / Accepted: 27 June 1997     

Adaptations to osmotic stress in the fresh-water euryhaline teleost Tilapia mossambica. IV. Changes in blood glucose,liver glycogen and muscle glycogen levels

Some aspects of osmoregulation energetics have been studied in the euryhaline teleost Tilapia mossambica (Peters) acclimated to media of different salinities. In stress media (75 and 100% sea water) the blood glucose of the fish increases significantly, accompanied by a corresponding increase in oxygen consumption and cytochrome-oxidase activity, suggesting that oxidative degradation of blood glucose is the predominant energy source for osmotic work in these stress media. It is likely that the variations in the blood-glucose level as a function of acclimation to the heterosmotic media —except the natural fresh-water medium — are governed by the combined effects of salinity of the medium and blood-medium osmotic gradient, rather than by the effect of any one of them separately. Perhaps, metabolic homoeostasis is in operation in the natural fresh-water medium. Depletion of muscle glycogen at significant levels is noticed only in stress media. Presumably, there is an augmentation of oxidative metabolism with glycolysis to meet the exacting energy demands for heavy osmotic work in high-stress media. Prior acclimation to 75% sea water (24.375 S) facilitates subsequent acclimation to 100% sea water (32.50 S) with less energy cost —an instance of facilitation acclimation. Smaller individuals of T. mossambica osmoregulate with less energy expenditure than larger ones. Thus, smaller individuals are osmotically more efficient.     

Normoxic and anoxic energy metabolism of the southern oyster drillThais haemastoma during salinity acclimation

The energetic cost associated with salinity acclimation was determined in the marine gastropodThais haemastoma by direct calorimetry under normoxic and anoxic conditions. Snails were collected from Caminada Pass near Grand Isle, Louisiana (Longitude 90°2W; Latitude 29°2N) in September 1987. Metabolic heat flux of snails acclimated to and measured at 10 or 30 S was similar at 15.06 or 16.39 J g^–1 dry wt h^–1, respectively, (corresponding to 0.76 or 0.83 ml O₂ g^–1 dry flesh wt h^–1) under normoxic conditions, and 2.39 or 2.53 J g^–1 dry wt h^–1 under anoxic conditions. Inter-individual variability was high, obscuring the effect of salinity gradient on heat flux. When standardized to the pre-transfer control level of each individual under anoxic conditions, a significant increase (55%) of energy expenditure was observed for snails transferred to hyperosmotic conditions. In contrast, heat flux varied insignificantly in individuals in the anoxic 30 to 10 S transfer. After transfer of individuals from 10 to 30 S under normoxic conditions, heat flux was depressed initially to 38% of the control rate, but recovered after 14 h to a higher metabolic rate (56%) than the pre-transfer control rate. After transfer of individuals from 30 to 10 S under normoxic conditions, the standardized heat flux decreased to 28% of the control rate, followed by a 20 h period of recovery to the control rate. The energy cost of intracellular hypoosmotic regulation was less than hyperosmotic regulation under anoxic conditions. The retraction of the foot ofT. haemastoma after normoxic salinity transfers did not generally correlate with the time course of metabolic heat flux.     

Long-term acclimation of the teleost Oreochromis mossambicus to various salinities: two different strategies in mastering hypertonic stress

Laboratory-reared tilapia (Oreochromis mossambicus) were long-term acclimated to freshwater (FW), brackish water (BW, 10 salinity), seawater (SW, 35 salinity) and two hypersaline media (45 and 60 salinity). We examined the influence of these ambient salinities on the density (D _cc) and diameter (d _cc) of DASPMI-stained chloride cells and on the capacity for electrogenic Cl^- secretion of the in vitro opercular epithelium. To provide a characterisation of Cl^- secretion, transepithelial potential difference (PD _te), conductance (G _te) and short-circuit current (I _sc) were measured after mounting the respective epithelium in an Ussing-chamber. The cellular electromotive forces (E _c) and conductances (G _c) as well as the leak conductances (G _l) were obtained from G _te: I _sc plots. In the salinity range between FW and SW both D _cc and d _cc increased. All electrophysiological parameters recorded increased in parallel, indicating a strong enhancement of the capacity for Cl^- secretion on the cellular and epithelial level. In the salinity range above SW a further increase of D _cc was observed. However, despite a higher concentration gradient across the body surface of the tilapia during acclimation to hypersaline media, the short-circuit current (I _sc) was not significantly different compared to SW preparations. This reflects proportional decreases of G _c and increases of E _c, respectively. Of particular interest, we found a strong decrease of the leak conductance (G _l) in preparations from tilapia acclimated to hypersaline media compared to those from SW fish, indicating that the tight junctions become less permeable.     

Phenotypic flexibility in response to environmental salinity in the euryhaline crab Neohelice granulata from the mudflat and the saltmarsh of a SW coastal lagoon

This study constitutes a first attempt to investigate intraspecific differences in osmoregulatory capacity and digestive and metabolic responses at the biochemical level in relation to hyper- and hypo-regulation in a single species of estuarine crab inhabiting contrasting habitats within a same intertidal area. We compared hemolymph osmolality, key digestive enzymes, glycemia and energy reserves in Neohelice granulata (Dana in Proc Acad Nat Sci Philadelphia 5:247–254, 1851) from the mudflat and saltmarsh of Mar Chiquita coastal lagoon (37°32′/37°45′S-57°19′/57°26′W) under a wide range of salinities (6–60 psu). Individuals from both sites exhibited high and similar osmoregulatory capacity, but while in individuals from mudflat low and high salinities affected lipase activity in hepatopancreas and triglycerides in muscle, in crabs from saltmarsh, high salinities affected glycogen in anterior gills. Low salinity differentially affected free glucose in anterior gills. The results suggest the occurrence of intraspecific distinct digestive and metabolic adjustments in relation to osmoregulatory responses and habitat.     

Effects of temperature and salinity acclimation of adults on larval survival,physiology, and early development of Lytechinus variegatus (Echinodermata: Echinoidea)

Larval survival and developmental rates of Lytechinus variegatus (Lamarck) were determined as a function of temperature and salinity in two experiments by: (1) directly transferring fertilized eggs to 35, 30, 27.5, 25, 20, 15, and 10S seawater at 18 and 23°C, and (2) acclimation of adult sea urchins to the conditions described above for 1 to 4 wk prior to spawning. Developmental rates and percent survival of larvae prior to metamorphosis decreased at salinities below 35 (Q₁₀ values for metamorphosis=0.380 to 0.384). Temperature and salinity significantly (P<0.05) affected metabolic rates of L. variegatus plutei. These results show that L. variegatus larvae are stenohaline when compared to larvae of other echinoderm species. LC₅₀ values (S), developmental rates, and survival to metamorphosis indicate that acclimation of adult sea urchins to lower salinity prior to spawing and fertilization does not enhance development or survival of embryos exposed to low salinity.     

Effect of salinity on oxygen consumption in postlarvae of the penaeid shrimps Penaeus monodon and P. stylirostris without and with acclimation

The effects of salinity on oxygen consumption was investigated in cultured Penaeus monodon Fabricius and P. stylirostris Stimpson postlarvae (32 and 35 d after metamorphosis, respectively). In both species, no difference appeared between non-acclimated and fully-acclimated individuals. The metabolic rate was unaffected by salinity variations, but P. stylirostris displayed a tendency (non-significant at P<0.05) to increase respiration at lower salinities. These observations are discussed in reference to the ecology of the young prawns.     

Some observations on the influence of salinity on growth and photosynthesis inPorphyra umbilicalis

Notwithstanding the great importance of the salinity factor in the marine environment, the knowledge of influence of salinity on growth of marine benthic algae is very limited. Rate of growth (mg, cm²) and O₂ output of the intertidal red algaPorphyra umbilicalis from Helgoland, North Sea, were measured during a 3 week culture in 3 different salinities (1/2-, 1- and 2-concentrated artificial sea water; Table 1). Under hypertonic conditions (2-concentrated sea water) growth rate and photosynthesis rate were depressed, compared to values obtained in normal concentrated sea water. Under hypotonic conditions (1/2-concentrated sea water), growth expressed in mg was the same as in normal concentrated sea water, or higher when expressed in cm³. Rate of O₂ output was almost unaltered in one of the two experiments, lowered in the other. Cell size increased at higher salinity, while swelling of cell walls and intercellular substances as well as the intensity of colouring decreased with salinity. The discrepancies between growth and photosynthesis under hypotonic conditions cannot be completely explained by the observed influences of salinity on morphological structures (cell size, swelling of cell substances). Detailed studies on the time course of photosynthesis and respiration rates, and preparation of a metabolic balance for the algae are necessary.     

Osmoregulation in the euryhaline flagellate Brachiomonas submarina (Chlorophyceae)

Brachiomonas submarina Bohlin (Chlorophyceae), a euryhaline marine flagellate, can osmoregulate over a wide range of external salinity. The alga exhibits maximum water content at 100% artificial seawater (ASW), and shows only a small water loss (<15%) when salinity is increased to 300% ASW. The non-aqueous volume of the cells is increased at salinities higher than 100% ASW. This is partially attributable to the accumulation of glycerol. Glycerol is the major osmoregulatory organic solute in this flagellate. The alga also shows an accumulation of amino acids in response to increased salinity. The contribution of glycerol and amino acids to intracellular osmolarity is only 9% at 10% ASW, but accounts for 49% at 300% ASW. The remainder of the osmotic balance is due to uptake and accumulation of inorganic ions, particularly sodium, potassium and chloride.     

Time-course of digestive-enzyme acclimation in the cockle Cerastoderma edule

We determined the temporal evolution of amylase, cellulase, laminarinase and protease in the digestive gland and crystalline style of cockles Cerastoderma edule held over 9 to 12 d in the presence and absence of food. Cockles were fed a constant diet of 1.5 mm³ l⁻¹ of Tetraselmis suecica for 9 to 12 d and were then starved for 6 to 8 d in late summer (September 1992) and in winter (January 1993). Feeding increased the dry weight and total cellulase, laminarinase and protease activities of the digestive gland irrespective of season, whereas amylase activity remained unchanged. In winter (i.e. when cockles are metabolically weak) the response was faster and stronger, especially for protease. An additional experiment in September starved cockles for 20 d before resuming feeding. In agreement with the seasonal differences, the presence of food after prolonged starvation induced a rapid and marked increase in protease in the digestive gland of the cockles. In winter, the possible effects of the biochemical composition of food on their enzymatic response were tested by feeding two groups of cockles with the same ration of T. suecica but harvested at different growth phases. A compensatory induction of cellulases occurred in cockles fed on T. suecica with a lower carbohydrate content. In the crystalline style, the protein level and carbohydrase fell during the first day of feeding and increased during the first day of subsequent starvation. These results indicate that the release of enzymes from the style prevails over the incorporation of enzymes during the early stages of feeding, whereas the opposite occurs during starvation. Received: 15 February 1998 / Accepted: 22 February 1999     

Carbofuran induced impairment in the hypothalamo-neurohypophyseal-gonadal complex in the teleost,Channa punctalus (Bloch)

The effect of chronic exposure to carbofuran (4.5 ppm in static water) for six months on the gonadal histophysiology and hypothalamo-neurohypophyseal complex was studied in Channa punctatus. Experimental observations revealed significant inhibition of gonadal development with associated degenerative abnormalities as evidenced by ovarian and testicular histology and reduced gonadosomatic index. Degenerative changes in ovary were exihibited by stage I (oogonium) and stage II (immature/non-vitellogenic) oocytes as marked by perinuclear ooplasmic lysis, clumping and dissolution resulting in disintigration of nuclear material altogether attributed to complete degeneration of such oocytes. Testicular deleterious changes included degeneration of spermatogenic elements and necrosis of interstitial cells of Leydig. Correlative histophysiological changes were also observed in the pituitary gonadotrophs and hypothalamic, nucleus pre-opticus, neurons that were smaller, inactive and less in number with associated necrosis. Corresponding to the changes in nucleus pre-opticus neurons, significant inhibition of brain monoamine oxidase enzyme activity was also recorded in treated group. These observations suggest that carbofuran even at low concentration level under long-term exposure is capable of inducing retardation of gonadal development which might have been mediated through the impairment of the hypothalamo-neurohypophyseal-gonadal axis in this species.     

Assimilation,inter-organ transfer and excretion of americium in two teleost fish

Radiotracer experiments were performed (February–April, 1982) to study the assimilation and metabolism of the transuranium nuclide americium-241 in the marine teleosts Serranus scriba (Linnaeus, 1758) and Scorpaena notata Rafinesque, 1810, caught off the Monaco coast. Fish fed with ²⁴¹Am-labelled food showed that assimilation of this radionuclide takes place through the gastrointestinal walls and that the small fraction accumulated is incorporated mainly in the skin, muscle and skeleton. Gut-transfer coefficients were similar in both species and averaged 0.7% (range 0.1 to 1.7%) of the ingested activity. The calculated biological half-lives for loss of the absorbed fraction ranged between 49 and 61 d for Serranus scriba and 12 and 117 d for Scorpaena notata. Results from an intramuscular injection experiment indicated that ²⁴¹Am was retained mainly in the liver, skin and skeleton; the fraction accumulated by muscle was very low. Liver displayed a relatively short biological half-time for ²⁴¹Am loss of roughly 24 d. Routes of ²⁴¹Am excretion from the teleosts appear to be through the kidneys, gills and feces with bile serving as a possible excretion route from the liver. From the limited amount of published information available for comparison, experimental evidence is presented which suggests that ²⁴¹Am taken up via the food chain is more biologically available to marine fish than is plutonium.     

Red blood cell osmotic fragility in the Spanish mackerel Scomberomorus maculatus

Red cell osmotic fragility was studied in 50 blood samples of the fish Scomberomorus maculatus (Mitchill). High levels of mean corpuscular fragility were found and hypotheses are presented to explain this phenomenon.     

Light acclimation states of phytoplankton in the Southern Ocean, determined using photosynthetic pigment distribution

In high-latitude waters such as the Southern Ocean, the primary production of phytoplankton supports the ecosystem. To understand the photo-acclimation strategy of such phytoplankton within cold environments, the vertical distribution profile of photosynthetic pigments was analyzed in the Southern Ocean. Samples were taken along 110°E during the austral summer, and along 150°E and around the edge of the seasonal sea ice of the Antarctic Continent during the austral autumn. Pigment extraction methods were optimized for these samples. The standing crop of chlorophyll a was larger in the region along the edge of the seasonal sea ice than at sampling stations in open ocean areas. Chlorophyll concentration seemed to be dependent on the formation of thermo- and haloclines along the edge of the seasonal sea ice, but not in the open ocean where such clines are less pronounced. The marker pigments fucoxanthin and/or 19′-hexanoyloxyfucoxanthin were dominant at most sampling stations throughout the water column, while other marker pigments such as alloxanthin were quite low. This indicated that diatoms and/or haptophytes were the major phytoplankton in this area. Comparison of the relative ratio of fucoxanthin with that of 19′-hexanoyloxyfucoxanthin allowed some stations to be characterized as either diatom-dominant or haptophyte-dominant. The relative ratio of xanthophyll-cycle pigments (diadinoxanthin plus diatoxanthin) to chlorophyll a was high in surface waters and decreased gradually with depth. This suggests that near the ice edge during summer in the Southern Ocean, both diatoms and haptophytes acclimate to their light environments to protect their photosystems under high-light conditions.     

A comparison of transport-related gill enzyme activities and tissue-specific free amino acid concentrations of Baltic Sea (brackish water) and freshwater threespine sticklebacks, Gasterosteus aculeatus, after salinity and temperature acclimation

The osmoregulatory abilities of one freshwater and two brackish water (Baltic Sea) populations of the euryhaline teleost fish Gasterosteus aculeatus were studied with respect to evolutionary physiology. Plasma osmolality, activities of Na⁺K⁺-ATPase, citrate synthase, creatine kinase in the gill and free amino acids in liver, axial muscle and pectoral fin muscle were measured. After transfer from 10 to 35 ppt at 15 °C, time-course changes of plasma osmolality and gill Na⁺K⁺-ATPase showed no significant fundamental differences between the freshwater and one of the Baltic Sea populations. In a multi-factorial experiment, each population was exposed to four different abiotic regimes. Both brackish water populations had high mortality in freshwater at 4 °C, which is discussed as a failure of osmotic regulation (reduced taurine concentrations). Freshwater specimens had higher levels of glycine in the axial and pectoral fin muscles compared to the brackish water populations. This is interpreted as a genetically based effect. In brackish (20 ppt) water of 15 °C, the freshwater population had high activities of Na⁺K⁺-ATPase, but low activities of creatine kinase, whereas both brackish water populations behaved in the opposite way. A fundamental difference between the freshwater and brackish water populations on the level of the osmoregulatory machinery was not observed. Received: 10 December 1998 / Accepted: 22 September 1999     

Different salinity tolerance mechanisms in Atlantic and Chesapeake Bay conspecific oysters: glycine betaine and amino acid pool variations

Crassostrea virginica (Gmelin) collected in 1989 from several sites within the Chesapeake Bay have narrower salinity tolerances than conspecific oysters collected in 1989 from several Atlantic coast sites (Georgia to Cape Cod). The basis of this physiological difference appears to be the biochemical mechanisms that control cellular osmolality following salinity stress. When adapted to the same salinity, the amino acid pools of both gill and adductor muscles of Atlantic oysters are larger than those of Bay oysters and different in composition. The Atlantic oyster tissues rely primarily on taurine for salinity tolerance, while the Bay oyster tissues have relatively less taurine, depending instead upon alanine, glycine and proline to adapt to high salinity. In addition, Atlantic oyster gill and adductor have 10 to 25 times the glycine betaine concentrations of these tissues from Bay oysters, depending upor the salinity of acclimation. The betaine concentration varies with salinity in Atlantic oysters, but does not change in Bay oysters. The results suggest that these biochemical differences are the basis of the narrower salinity tolerance in Bay oysters. The biochemical differences may reflect genetic differences between Bay and Atlantic oysters.     

Metabolic constraints on burst-swimming in the Antarctic teleost Notothenia neglecta

Immediate-energy-supply pathways were analysed during burst-swimming in white muscle of the Antarctic fish Notothenia neglecta, (collected during Austral summer 1984 near Signy Island, South Orkneys). Selected enzyme activities were determined, and tissue metabolites were measured, before and after 3 min of burst-work. The activity of glycolytic enzymes is very low compared to that in other fish, while the activities of creatine phosphokinase, adenylate kinase and AMPdeaminase are relatively high. Concentrations of metabolites associated with cellular energy status declined with exercise. At the same time, there was no change in the concentrations of most of the glycolytic metabolites measured (including glycogen, glucose and lactate), while [creatine phosphate] declined and [inosine monophosphate] increased. These observations were used to propose that the potential for ATP generation via creatine-phosphate hydrolysis is enhanced in N. neglecta, while glycolytic capacity is relatively reduced. This is an unusual pattern of ATP production and places severe constraints upon the behavior of the fish. While the capacity for very short-term, high-power-output swimming is increased, the capacity for medium-duration swimming is reduced. Ecological repercussions are discussed.