Influence of environmental salinity on oxygen consumption and ammonia excretion of the arctic under-ice amphipodOnisimus glacialis

Changes in salinity affect the metabolic rate of the sympagic amphipodOnisimus glacialis collected from the Barents Sea in 1986 and 1988. When transferred from 35 to 5 ppt S, oxygen consumption and ammonia excretion both increase three-fold during the first 5 h of exposure, and they remain high throughout the rest of the experimental period (26 h). During 24-h acclimation to various salinities (5 to 45 ppt), the amphipods exhibit a respiratory and excretory response to hyper- and hypoosmotic stress; however, a rather constant O:N atomic ratio (around 15) was obtained at the experimental salinities, indicating protein/lipids as metabolic substrate. Both rates of oxygen consumption and ammonia excretion increased with an increasing osmotic difference (0 to 650 mOsm) between the haemolymph and the environmental medium, indicating higher energy requirements for osmotic and ionic regulation at low salinities. In amphipods abruptly transferred from 35 to 5 ppt, a minor decrease of the haemolymph sodium concentrations together with an increased ammonia excretion output indicate a counter-ion regulation of NH ₄ ⁺ and Na⁺ during hyposmotic stress.     

Tolerance of Mediterranean seagrasses (<Emphasis Type="Italic">Posidonia oceanica</Emphasis> and <Emphasis Type="Italic">Cymodocea nodosa</Emphasis>) to hypersaline stress: water relations and osmolyte concentrations

The present study examines for the first time the effects of increased salinity on water relations and osmolyte (carbohydrates and amino acids) concentrations in two Mediterranean seagrass species, Posidonia oceanica and Cymodocea nodosa, which are adapted to growth in environments with contrasting salinity and have a known differential sensitivity to alterations in ambient salinity. The specific aim was to obtain insights into their respective capacities to cope with natural or anthropogenically induced (e.g. desalination plants) hypersaline stress and its ecological implications. To this end, large plant fragments of both seagrass species were maintained for 47 days in a laboratory mesocosm system under ambient salinity (37 psu; control) and three chronic hypersaline conditions (39, 41 and 43 psu). Analyses of leaf-tissue osmolality indicated that both species followed a dehydration avoidance strategy, decreasing their leaf water potential (Ψ_w) as the external salinity increased, but using different physiological mechanisms: whereas P. oceanica leaves exhibited a reduction in osmotic potential (Ψ_π), C. nodosa leaves maintained osmotic stability through a decrease in turgor pressure (Ψ_p) probably mediated through cell-hardening processes. Accordingly, the concentrations of soluble sugars and some amino acids (mainly Pro and Gly) suggested the activation of osmoregulatory processes in P. oceanica leaves, but not in C. nodosa leaves. Osmotic adjustments probably interfered with leaf growth and shoot survival of P. oceanica under hypersaline stress, whereas C. nodosa showed a more efficient physiological capacity to maintain plant performance under the same experimental conditions. These results are consistent with the more euryhaline ecological behaviour of C. nodosa and contribute to understanding the high vulnerability shown by P. oceanica to even mild increments in seawater salinity.     

Upper temperature tolerances of some European molluscs. I Tellina fabula and T. tenuis

The upper temperature tolerances of two European species of Tellina from populations near the northern and southern limits of their geographical range are compared by means of median lethal temperature (LT₅₀), and median burial temperature (BT₅₀) determination for periods of exposure up to 96 h. The influence of previous acclimation temperature, and salinity within a limited range, on these determinations is also considered. T. fabula Gmelin, which is found in deeper water throughout its range, shows a lower thermal tolerance than T. tenuis da Costa. Both species display a significant effect of previous acclimation temperatures, the effect on the burrowing responses being greater than that shown by LT₅₀, and in both species the thermal tolerance of individuals from Mediterranean populations is greater than that of individuals from North Atlantic populations.     

Pre-immersion salinity-choice behaviour in Porcellana platycheles

The ability of the littoral porcelain crab Porcellana platycheles (Pennant) to recognise and avoid low-salinity levels when immersed in media has already been demonstrated in previous experiments. It was thought that the ability to identify and avoid diluted sea water before complete immersion would also be of survival value, since this would prevent exposure of the permeable body surfaces (especially the gills) to the swelling effects of the osmotic uptake of water. The experiments reported here demonstrate that such pre-immersion salinity choice behaviour may be elicited from P. platycheles in the laboratory, and that salinity reception is apparently sited upon the walking limbs. Choice between salinity levels is shown to be based on the osmotic concentration of media rather than on the concentration of a particular ion or group of ions.     

Interaction of short-term testosterone treatment with osmotic acclimation in the gilthead sea bream <Emphasis Type="Italic">Sparus auratus</Emphasis>

To assess the interaction between testosterone (T) treatment and acclimation to different salinities, seawater-acclimated gilthead sea bream (Sparus auratus) were implanted with slow-release coconut oil implants alone (control) or containing T (5 μg/g body mass). After 5 days, eight fish of control and T-treated groups were sampled. The same day, eight fish of each group were transferred to low salinity water (LSW, 6 ppt, hypoosmotic test), seawater (SW, 38 ppt, control test) and high salinity water (HSW, 55 ppt, hyperosmotic test) and sampled 9 days later. Gill Na⁺, K⁺-ATPase activity increased in HSW-acclimated fish with respect to SW- and LSW-acclimated fish in both control and T-treated groups. Kidney Na⁺, K⁺-ATPase activity was also enhanced in HSW-acclimated fish, but only in T-treated group. From a metabolic point of view, most of the changes observed can be attributed to the action of salinity and T treatment alone, since few interactions between T treatment and osmotic acclimation to different salinities were observed. Those interactions included in treated fish: in the liver, decreased capacity in using glucose in fish acclimated to extreme salinities; in the gills, decreased capacity in using amino acids in HSW; in the kidneys increased capacity in using amino acids in extreme salinities; and in the brain, decreased glycogen and acetoacetate levels of fish in LSW.     

Effect of environmental parameters on lysosomal marker enzymes in the tropical blood clam Anadara granosa

The lysosomal marker enzymes, arylsulfatase and acid phosphatase, in a tropical burrowing arcid clam Anadara granosa L. have been found to exhibit seasonal variations. The activity of both enzymes decreased with increase in ambient temperature and fell with increase in salinity. Lysosomal latency for these enzymes, however, was not significantly affected by environmental parameters, including salinity, temperature, nutritional status, breeding season, etc. The physico-chemical characteristics of the habitat were found to induce reversible changes in the lysosomal latency and enzyme activity. Exposure to elevated temperature (T=7° to 8°C), towards the upper limit of its physiological tolerance, was found to labilize the lysosomes. The biochemical methods optimized for the demonstration of lysosomal stability under exposure to environmental stressors could also be extended to quantify the impact of various pollutants, including heavy metals, radionuclides, etc. at the subcellular level.     

Synergistic effects of temperature,salinity and light on the hermatypic coral Montipora verrucosa

Temperature tolerance in the reef coral Montipora verrucosa (Lamarck) is affected by salinity and light. Low salinity reduces ability of the coral to survive shortterm exposure to elevated temperature. High natural light intensity aggravates damage sustained by corals at high temperature. In long-term growth experiments, high light intensity caused substantial loss of zooxanthellar pigment, higher mortality rates, reduced carbon fixation and lowered growth rate at both upper and lower sublethal temperatures Effects of light at optimal temperature were less dramatic. Interactions between physical environmental factors appear to be most important near the limits of tolerance for a given factor. Acclimation capability was indicated, and was influenced by both thermal history and pigmentation state of stressed corals.Contribution No. 543 of the Hawaii Institute of Marine Biology.     

Temporal and spatial distribution of three supralittoral amphipod species on a sandy beach of central Italy

The distribution of three talitrid species—Talitrus saltator (Montagu, 1808), Orchestia gammarella (Pallas, 1766), Platorchestia platensis (Kroyer, 1845)—in the beach-dune system at the mouth of the Mignone River (central Italy) was analysed. It was related to the variations of the following abiotic factors: temperature, penetrability, pH, conductivity and moisture of the sediment. The beach-dune system is influenced by human impact and natural erosion. All species showed a maximum capture frequency in November, while the abundance decreased to a minimum in July. T. saltator was dominant on the beach, the other two along the riverbank. Juveniles were abundant along the riverbank; they were most abundant in November and almost disappeared in summer. T. saltator was more abundant near the waterline during the hottest months and occupied the inner beach in winter, with occasional presences on the dune. Regression analysis between the abiotic factors and species abundance showed a positive relationship with pH and temperature for T. saltator, while O. gammarella was negatively related to pH. Canonical correspondence analysis (CCA) showed that penetrability, moisture and temperature had the greatest influence on the species. T. saltator was almost entirely confined to the beach transects and strongly associated with penetrability, temperature and pH values. O. gammarella was mostly associated with the riverbank and P. platensis only found there and on a nearby pool. The two species were mainly influenced by moisture and variations in grain sizes of the sediment.     

Genetic differentiation between populations of Talitrus saltator and Talorchestia deshayesii (Crustacea: Amphipoda) from coastal areas of the north-western European continent

Electrophoretic studies of gene-enzyme variation in the littoral talitrids Talitrus saltator (Montagu) and Talorchestia deshayesii (Audouin) were undertaken to estimate the amount of divergence among geographically separated populations. Samples of both species were taken from sandy beaches over a transect of approximately 3 500 km along the coast of the European continent including Baltic, North Sea and Atlantic locations. A total of 22 T. saltator and 15 T. deshayesii populations were analysed for genetic variation at various enzyme loci. Both amphipods revealed relatively low levels of polymorphism and heterozygosity. Among the loci studied, phosphoglucose isomerase (Pgi) and phosphoglucomutase (Pgm) were highly polymorphic. Patterns of micro- and macrogeographic variation in terms of distributions of allele frequencies at these particular loci are compared. Interpopulation allozymic variation was shown to be lower in T. deshayesii than in T. saltator. As demonstrated by T. saltator populations sampled in coastal sites ranging from Denmark to western France, clinal variation in frequencies of two alleles became evident at the PGI locus, exhibiting a steady increase in the level of polymorphism with decreasing latitudes. It is argued that limited gene flow and, to some extent, random genetic drift may account for the gene pool structure of the talitrid species investigated.     

Dopamine,noradrenaline and serotonin during hypo-osmotic stress of Carcinus maenas

Dopamine, noradrenaline and serotonin were measured in the haemolymph and gills of Carcinus maenas before and after transferring them into diluted seawater. A remarkable variation of the above-mentioned compounds was observed as a function of time of osmotic stress. C. maenas were captured in the Venetian lagoon near Chioggia, Italy in 1986.     

Effects of salinity on the clam <Emphasis Type="Italic">Chamelea gallina</Emphasis> haemocytes. Part II: Superoxide dismutase response

The effects of different salinity levels (28, 34 and 40‰) on functional responses of Chamelea gallina haemocytes were evaluated in a two part study dealing with modulations of immune parameters. This part (Part II) of the study was focused on the superoxide dismutase (SODs) activity and expression in haemocyte lysate and cell-free haemolymph. Results of this study established that the exposure of C. gallina specimens at 40‰ salinity provoked a decrease in Mn-SOD and Cu/Zn-SOD activities in haemocyte lysate suggesting a declining superoxide anion generation at the highest salinity tested. Expression of MnSOD was coherent with activity values, while Cu/ZnSOD showed two immunoreactive bands. The former corresponds to the cytosolic Cu/Zn-SOD (16 kDa) was not coherent with the enzyme activity and the second (28–30 kDa) probably attributed to EC-SOD. In cell-free haemolymph, Mn-SOD activity decrease and Cu/Zn-SOD activity increase at 40‰ were observed, likely due to EC-SOD contribution, strongly induced at the same salinity. After EC-SOD detection with two different antibody, we postulate that the EC-SOD like-protein band (29 kDa) may be constitute partly by EC-SOD and probably by Cu/Zn-SOD dimeric form not completely dissociated under reducing condition or a Cu/Zn-SOD degenerated but still recognized by antibody. The cell-free haemolymph increase of EC-SOD at high salinity values plays an important role in immune defence of C. gallina. According to the conclusion of Part I of this study, our data confirmed the destabilizing effect of 40‰ salinity on haemocyte functionality, while to 28‰ exposure, data don’t confirm its stressful action as instead stated by results of Part I. Further studies are necessary to clear up this discrepancy.     

Metabolic response under different salinity and temperature conditions for glass eel Anguilla japonica

Diadromous fish often enter freshwater directly from seawater via fish ladders or channels built in estuarine dams. The oxygen consumption rates (OCR) of glass eel, Anguilla japonica, were determined using an automatic intermittent flow respirometer under various salinity and temperature regimes to physiologically explain this direct movement. The endogenous rhythm of the OCR in wild glass eels, freshly collected from estuaries, was nearly synchronous with the tidal pattern at the estuarine collection site. When the salinity was changed from 20 psu (12°C) at a constant temperature to that of freshwater, the OCR of the glass eels decreased by 21.6±7.0% (mean ± SD) (P<0.05), showing a dampened rhythm for about 48 h. After this period of impediment, the glass eels resumed normal metabolic activity. Direct migration from seawater to freshwater under constant temperature would result in a severe physiological stress for these glass eels for about two days. When the glass eels were exposed to a cyclic change in water temperature of 2°C 26 h⁻¹, as they encounter in estuaries, and then were introduced to freshwater abruptly, the OCR rhythm corresponded to the cyclic changes in water temperature after exposure to freshwater. Under these conditions, the mean OCR of the glass eels had a small difference before and after exposure to freshwater. These data explained how glass eels can directly move from sea water into the freshwater without any apparent metabolic stress in the estuaries showing cyclic change in water temperature (Δt=2°C).     

The synergistic effects of increasing temperature and CO2 levels on activity capacity and acid–base balance in the spider crab, Hyas araneus

With global climate change, ocean warming and acidification occur concomitantly. In this study, we tested the hypothesis that increasing CO₂ levels affect the acid–base balance and reduce the activity capacity of the Arctic spider crab Hyas araneus, especially at the limits of thermal tolerance. Crabs were acclimated to projected oceanic CO₂ levels for 12 days (today: 380, towards the year 2100: 750 and 1,120 and beyond: 3,000 μatm) and at two temperatures (1 and 4 °C). Effects of these treatments on the righting response (RR) were determined (1) at acclimation temperatures followed by (2) righting when exposed to an additional acute (15 min) heat stress at 12 °C. Prior to (resting) and after the consecutive stresses of combined righting activity and heat exposure, acid–base status and lactate contents were measured in the haemolymph. Under resting conditions, CO₂ caused a decrease in haemolymph pH and an increase in oxygen partial pressure. Despite some buffering via an accumulation of bicarbonate, the extracellular acidosis remained uncompensated at 1 °C, a trend exacerbated when animals were acclimated to 4 °C. The additional combined exposure to activity and heat had only a slight effect on blood gas and acid–base status. Righting activity in all crabs incubated at 1 and 4 °C was unaffected by elevated CO₂ levels or acute heat stress but was significantly reduced when both stressors acted synergistically. This impact was much stronger in the group acclimated at 1 °C where some individuals acclimated to high CO₂ levels stopped responding. Lactate only accumulated in the haemolymph after combined righting and heat stress. In the group acclimated to 1 °C, lactate content was highest under normocapnia and lowest at the highest CO₂ level in line with the finding that RR was largely reduced. In crabs acclimated to 4 °C, the RR was less affected by CO₂ such that activity caused lactate to increase with rising CO₂ levels. In line with the concept of oxygen and capacity limited thermal tolerance, all animals exposed to temperature extremes displayed a reduction in scope for performance, a trend exacerbated by increasing CO₂ levels. Additionally, the differences seen between cold- and warm-acclimated H. araneus after heat stress indicate that a small shift to higher acclimation temperatures also alleviates the response to temperature extremes, indicating a shift in the thermal tolerance window which reduces susceptibility to additional CO₂ exposure.     

Aspects of nitrogen metabolism of the common mussel Mytilus edulis: Adaptation to abrupt and fluctuating changes in salinity

Mytilus edulis L. were exposed to abrupt (3015 and 1530) and fluctuating (sinusoidal 12 h cycles of 301530) changes in salinity, and the changes in the total osmoconcentration of the haemolymph were recorded. The response of nitrogen metabolism to the altered extracellular osmotic concentrations was investigated in terms of the concentrations of the total NPS (ninhydrin-positive substances) pool and the individual amino acids of the tissues, the concentration of the amino acids of the haemolymph, and the rates of excretion of ammonia and amino acids by whole individuals. The haemolymph became isosmotic with the seawater with abrupt changes in salinity, but with fluctuating salinity was slightly hyperosmotic as the salinity decreased and then slightly hypo-osmotic as the salinity increased. This resulted in a reduction in the extent of the extracellular osmotic change compared to the change in fluctuating salinity to which it was exposed. Total NPS of the tissues decreased with an abrupt decrease in salinity and increased with an abrupt increase in salinity, but a seasonal dependence of the response was indicated. The short-term response of tissue NPS to fluctuating salinity was equivocal, but with long-term exposure the concentration declined. Ammonia and amino acid excretion increased with both an abrupt decrease in salinity and fluctuating salinity and decreased with an abrupt increase in salinity. Haemolymph amino acids increased with an abrupt decrease in salinity. The increased rates of nitrogen excretion accounted for the reductions in the NPS concentrations of the tissues except in the early stages of fluctuating salinity. Taurine, aspartate, threonine, serine, glycine and arginine declined with an abrupt decrease in salinity while alanine and glutamate increased slightly. With an abrupt increase in salinity, alanine and ammonia accumulated in the tissues and then declined while the other amino acids increased slowly over a longer time-course. Similar individual amino acid responses were seen with long-term exposure to fluctuating salinity, except for taurine which did not decrease in concentration. On the basis of the changes in tissue amino acids and ammonia, it is suggested that the alanine dehydrogenase reaction is the primary nitrogen-fixing reaction in marine bivalves such as M. edulis.     

Heavy metal contamination and physiological variability in the Brazilian mangrove crabs Ucides cordatus and Callinectes danae (Crustacea: Decapoda)

 Physiological studies were made on the crabs Ucides cordatus (L.) and Callinectes danae sampled from populations living in “polluted” mangroves on the southeast littoral of Brazil. Analysis of Cu, Cd, Zn, and Fe of sediments and crab tissues showed interspecific differences in tissue concentrations, and significantly higher levels of Cu, Cd, and Zn in “polluted” populations compared to “unpolluted” crabs living in uncontaminated mangrove in the same geographical area. Individuals of both species from the polluted site showed significantly greater capacities for regulating blood osmotic concentrations at low salinity (9‰). However, U. cordatus showed a reduced hypo-regulatory ability in 34‰S. Differences in ionoregulation were also seen. “Polluted”C. danae showed significantly higher Na/ K-ATPase levels in posterior gills compared to “unpolluted” crabs. Oxygen consumption rates (M˙ _O2) were elevated in U. cordatus, but depressed in C. danae from the “polluted” population. Individuals of both species from this site showed significantly lower O:N ratios, mainly because of an increased net efflux of ammonia. Adenylate energy charge (AEC) values of muscle and hepatopancreas in “unpolluted” and “polluted” populations of both species were not significantly different. These physiological differences are discussed in relation to the known acute physiological and metabolic effects of heavy metals in crustaceans, and interpretated in the light of possible adaptive changes following long-term exposure to contamination. Received: 6 August 1999 / Accepted: 22 June 2000     

Effects of fluctuating salinity on the behaviour of the West African blood clam Anadara senilis and on the osmotic pressure and lonic concentrations of the haemolymph

Specimens of the West African blood clam Anadara senilis were exposed to both sinusoidal and abrupt salinity regimes. Measurements showed that haemolymph osmolality and concentrations of Na, K⁺, Ca²⁺ and Mg²⁺ followed the concentrations of the external medium up to the time of shell valve closure. Shell valves closed when the seawater concentration had fallen to about 15.4% S and reopened at a similar salinity. The closure mechanism was effective in preventing excessive haemolymph dilution.     

A neutral DNA marker suggests that parallel physiological adaptations to open shore and salt marsh habitats have evolved more than once within two different species of gastropods

Local adaptation is an important mechanism generating physiological diversity and can be especially pronounced in species with restricted dispersal and gene flow such as direct developing snails of the genus Littorina. We compared physiological responses to salinity and desiccation stress in two co-occurring species of northeastern Pacific Littorina (L. subrotundata and L. sitkana) with salt marsh and open shore ecotypes. The animals from salt marsh populations were significantly more tolerant to low salinities and significantly less resistant to desiccation stress than their open shore counterparts. The lower resistance to desiccation in salt marsh animals was not associated with a higher rate of water loss during air exposure or with lower body water reserves, but instead reflected a lower tolerance to high salinities. These habitat-related physiological differences occurred in parallel in the two studied species of Littorina and persisted after prolonged laboratory acclimation, suggesting that they may reflect selection for markedly different local optima in the salt marsh habitats than in the open shore habitats. We used a neutral polymorphic nuclear DNA marker (intron of aminopeptidase N) to estimate the level of gene flow between the populations from different habitats and found isolation by distance regardless of the habitat from which the snails were collected. Our molecular data suggest that physiological cohesiveness of ecotypes can arise despite different genetic backgrounds, and could potentially be due to parallel evolution of convergent phenotypes in similar habitats.Communicated by O. Kinne, Oldendorf/Luhe     

Oxygen bioavailability and haemoglobins in the brine shrimp Artemia franciscana

Bioavailability of oxygen for the saline-water invertebrate Artemia franciscana was studied, since both oxygen concentration and oxygen diffusion rate change with salinity. Total haemoglobin concentration and the relative contribution of each of three haemoglobins was measured in specimens acclimated to different salinities and oxygen concentrations. Both haemoglobin concentration and contribution were influenced by salinity and the group observed (males, females with and females without eggs). Multiple regression analysis showed that total haemoglobin concentration was better correlated with oxygen concentration than oxygen partial pressure. The relative proportions of haemoglobins 1, 2 and 3 were better correlated with oxygen partial pressure than oxygen concentration. These results are related to the oxygen carrying capacity and oxygen affinity of the haemolymph. Our results show that oxygen bioavailability in a saline-water environment depends on the response (relative contribution of Hb1, Hb2 and Hb3 or total haemoglobin concentration) that is studied. They also show that oxygen concentration and oxygen partial pressure have a different physiological impact on brine shrimp.