Branchial carbonic anhydrase (CA) of gills of Chasmagnathus granulata (Crustacea Decapoda)

The occurrence, localization and response to environmental salinity of carbonic anhydrase (CA) activity were studied in all of the gills of the euryhaline crab Chasmagnathus granulata from Mar Chiquita coastal lagoon (Buenos Aires Province, Argentina). CA activity in all gills appeared to be dependent on salinity. The pattern of distribution of CA activity among gills was different upon transition of C. granulata from osmoionoconformity (more uniform distribution) to hyperregulation (highest activity in posterior gills 6-8). Upon abrupt salinity change a differential response of CA activity occurred among gills which could suggest a differential role of CA in ion transport process in different gills of this crab. Furthermore, CA activity in anterior and posterior gills was found in cytosolic and microsomal fractions, although highest activity appeared to be membrane-associated. Both pools of CA were also strongly influenced by salinity and very sensitive to sulfonamide acetazolamide. The results suggest a differential participation of branchial CA in ionoregulatory mechanisms of C. granulata.     

Effects of environmental anoxia and different periods of reoxygenation on oxidative balance in gills of the estuarine crab Chasmagnathus granulata

We investigated the effects of anoxia (8 h) and different periods of reoxygenation (20 and 40 min) on the oxidative balance in anterior and posterior gills of the crab Chasmagnathus granulata. Enzyme activity of catalase and GST was increased in the gills of the animals submitted to anoxia, and SOD activity was decreased. These enzymes returned approximately to control levels during the anoxia recovery time. These results demonstrated enzyme activities change with variations in environmental oxygen levels. The posterior gills showed a higher antioxidant enzyme activity than anterior gills. In the gills, there were no changes in the non-enzymatic antioxidant system (TRAP) during anoxia. On the other hand, during anoxia recovery, an increase of TRAP in both gills was observed. Anoxia does not change lipid peroxidation (TBARS) in the gills. During anoxia recuperation, an increase in levels of TBARS was observed. Thus the results demonstrate that C. granulata has a similar strategy of preparation for oxidative stress as observed in other intertidal species, enabling the crabs to survive in an environment with extreme variations in physical and chemical characteristics, such as salt marshes.     

Salinity dependent Na+-K+ATPase activity in gills of the euryhaline crab Chasmagnathus granulata

The occurrence and response of Na+-K+ATPase specific activity to environmental salinity changes were studied in gill extracts of all of the gills of the euryhaline crab Chasmagnathus granulata from Mar Chiquita coastal lagoon (Buenos Aires Province, Argentina). All of the gills exhibited a salinity dependent Na+-K+ATPase activity, although the pattern of response to environmental salinity was different among gills. As described in other euryhaline crabs highest Na+-K+ATPase specific activity was found in posterior gills (6 to 8), which, with exception of gill 6, increased upon acclimation to reduced salinity. However, a high increase of activity also occurred in anterior gills (1 to 5) in diluted media. Furthermore, both short and long term differential changes of Na+-K+ATPase activity occurred among the gills after the transfer of crabs to reduced salinity. The fact that variations of Na+-K+ATPase activity in the gills were concomitant with the transition from osmoconformity to ionoregulation suggests that this enzyme is a component of the branchial ionoregulatory mechanisms at the biochemical level in this crab.     

Lipids as energy source during salinity acclimation in the euryhaline crab Chasmagnathus granulata dana, 1851 (crustacea-grapsidae)

Lipids seem to be the major energy store in crustaceans. Moreover, they are extremely important in maintaining structural and physiological integrity of cellular and sub cellular membranes. During salinity adaptation, energy-demanding mechanisms for hemolymph osmotic and ionic regulation are activated. Thus, the main goal of this work was to verify the possible involvement of lipids as an energy source in the osmotic adaptation process. The estuarine crab Chasmagnathus granulata was captured and acclimated to salt water at 20 per thousand salinity and 20 +/- 2 degrees C, for 30 days. After acclimation, crabs were divided into groups of ten and transferred to fresh water (0 per thousand ), salt water at 40 per thousand salinity, or maintained in salt water at 20 per thousand salinity (control group), without feeding. Before and seven days after the salinity change, wet weight and lipid concentration in gills, muscle, hepatopancreas, and hemolymph were determined according to the colorimetric assay of sulphophosphovanilin. Results show that hepatopancreas lipids were not mobilized during osmotic stress regulation. Gill and muscle lipids were significantly lower in crabs subjected to hypo-osmotic stress than those subjected to the hyper-osmotic stress or maintained at the control salinity. Our results point to the occurrence of lipid mobilization and involvement of these compounds in the osmotic acclimation process in C. granulata, but with differences between tissues and the osmotic shock (hypo or hyper) considered.     

Gill Na(+),K(+)-ATPase and osmoregulation in the estuarine crab, Chasmagnathus granulata Dana, 1851 (Decapoda, Grapsidae)

Some kinetic properties of gill Na(+),K(+)-ATPase of the estuarine crab, Chasmagnathus granulata, and its involvement in osmotic adaptation were analyzed. Results suggest the presence of different Na(+),K(+)-ATPase isoforms in anterior and posterior gills. They have different affinities for Na(+), but similar affinity values for K(+), Mg(2+), ATP and similar enzymatic profiles as a function of temperature of the incubation medium. Ouabain concentrations which inhibit 50% of enzyme activity were also similar in the two types of gills. Enzyme activity and affinity for Na(+) are higher in posterior gills than in anterior ones. Furthermore, affinities of Na(+),K(+)-ATPase of posterior gills for Na(+) and K(+) were similar to or higher than those of gills or other structures involved in the osmoregulation in several euryaline decapod crustaceans. Acclimation to low salinity was related to a significant increase in the maximum Na(+), K(+)-ATPase activity, mainly in posterior gills. On the other hand, crab acclimation to high salinity induced a significant decrease in maximum enzyme activity, both in anterior and posterior gills. These results are in accordance to the osmoregulatory performance showed by C. granulata in diluted media, and point out the major role of posterior gills in the osmoregulation of this species.     

Between-habitat comparison of digestive enzymes activities and energy reserves in the SW Atlantic euryhaline burrowing crab Neohelice granulata

The digestive and metabolic characteristics at the biochemical level underlying between-habitat dietary shift of the SW Atlantic euryhaline burrowing crab Neohelice granulata under natural conditions are unknown. We made studies on adult males of N. granulata from the open mudflat and the vegetated saltmarsh in a SW Atlantic costal lagoon (Mar Chiquita, 37°32'-37°45'S; 57°19'-57°26'W, Argentina). We determined and compared amylase, maltase, sucrase, proteolytic, lipase and alkaline phosphatases activities in the hepatopancreas; glycemia, and glycogen, free glucose, triglycerides and protein concentrations in hepatopancreas, chela muscle, and anterior and posterior gills. The results show that N. granulata exhibits characteristics and between-habitat differences at the biochemical level (i.e. high amylase and disaccharidase activities, differences in total proteolytic, lipase and levamisole-insensitive AP activities in the hepatopancreas, and in the concentrations of glycogen in the gills, triglycerides in the hepatopancreas and of protein in the chela muscle) which could represent adaptive digestive and metabolic strategies to face the differences in environmental conditions (i.e. food availability). The possible relationship between digestive and metabolic characteristics and feeding patterns, type of food available and environmental conditions in each habitat is discussed.     

Testing the effect–response framework: key response and effect traits determining above‐ground biomass of salt marshes

Question: How do species traits respond to environmental conditions and what is their effect on ecosystem properties? Location: Salt marshes, Northwest Germany. Methods: On 113 plots along the German mainland coast and on one island, we measured environmental parameters (soil nutrient content, inundation frequency, groundwater level and salinity), collected traits from 242 individuals (specific leaf area [SLA], whole plant C:N ratio, and dry weights of plant organs) and sampled above‐ground biomass as an ecosystem property. We constructed a path model combining environmental parameters, functional traits at community level and above‐ground biomass, which was tested against a dependence model using path analysis; model fit was evaluated by structural equation modelling (SEM). Results: The final model showed good consistency with the data and highlights the major role of groundwater level, salinity and nutrient availability as the most important factors influencing biomass allocation in salt marshes. Above‐ground living biomass was mostly determined by stem biomass, which was mediated through an allometric allocation of biomass to all other plant organs, including leaf mass. C:N ratio and SLA were the major drivers for dead biomass. Conclusion: We emphasize an indirect link between standing biomass and environmental conditions and recognize stem biomass, plant C:N ratio and SLA as keystone markers of species functioning in determining the relationship between environment and ecosystem properties.     

Effects of photoperiod on gluconeogenic activity and total lipid concentration in organs of crabs, Neohelice granulata, challenged by salinity changes

Abstract. This study assessed the effects of long (LD) or short (SD) days on the conversion of [¹⁴C]-glycerol to [¹⁴C]-glucose and total lipid concentration in organs of the crab Neohelice granulata challenged by a change in external salinity. In the 20‰-acclimated crabs, no difference was found in the concentration of total lipids in the muscle, hepatopancreas, gills, or hemolymph between crabs acclimated to SD or LD. In SD crabs, the total lipid levels in the anterior and posterior gills did not decrease during an osmotic challenge. Only in the posterior gills did the total lipid levels decrease during acclimation to the 34‰ medium in LD animals. The total lipid concentration in the hemolymph decreased after 1 d of osmotic stress in SD, and increased in the hepatopancreas. In LD crabs, the lipid contents decreased gradually in muscle, and in the hepatopancreas on day 3 after transfer to 34‰ medium. In 20‰-acclimated crabs, the gluconeogenesis activity in both sets of gills was higher in LD than in SD animals. The gluconeogenesis capacity decreased in both sets of gills on the first day of osmotic challenge in SD, and in the posterior gills on the third day in LD crabs. These results suggest that in organs of N. granulata , photoperiod affects the metabolic adjustments to an osmotic challenge.     

Copper effects on key metabolic enzymes and mitochondrial membrane potential in gills of the estuarine crab Neohelice granulata at different salinities

The estuarine crab Neohelice granulata was exposed (96h) to a sublethal copper concentration under two different physiological conditions (hyperosmoregulating crabs: 2ppt salinity, 1mg Cu/L; isosmotic crabs: 30ppt salinity, 5mg Cu/L). After exposure, gills (anterior and posterior) were dissected and activities of enzymes involved in glycolysis (hexokinase, phosphofructokinase, pyruvate kinase, lactate dehydrogenase), Krebs cycle (citrate synthase), and mitochondrial electron transport chain (cytochrome c oxidase) were analyzed. Membrane potential of mitochondria isolated from anterior and posterior gill cells was also evaluated. In anterior gills of crabs acclimated to 2ppt salinity, copper exposure inhibited hexokinase, phosphofructokinase, pyruvate kinase, and citrate synthase activity, increased lactate dehydrogenase activity, and reduced the mitochondrial membrane potential. In posterior gills, copper inhibited hexokinase and pyruvate kinase activity, and increased citrate synthase activity. In anterior gills of crabs acclimated to 30ppt salinity, copper exposure inhibited phosphofructokinase and citrate synthase activity, and increased hexokinase activity. In posterior gills, copper inhibited phosphofructokinase and pyruvate kinase activity, and increased hexokinase and lactate dehydrogenase activity. Copper did not affect cytochrome c oxidase activity in either anterior or posterior gills of crabs acclimated to 2 and 30ppt salinity. These findings indicate that exposure to a sublethal copper concentration affects the activity of enzymes involved in glycolysis and Krebs cycle, especially in anterior (respiratory) gills of hyperosmoregulating crabs. Changes observed indicate a switch from aerobic to anaerobic metabolism, characterizing a situation of functional hypoxia. In this case, reduced mitochondrial membrane potential would suggest a decrease in ATP production. Although gills of isosmotic crabs were also affected by copper exposure, changes observed suggest no impact in the overall tissue ATP production. Also, findings suggest that copper exposure would stimulate the pentose phosphate pathway to support the antioxidant system requirements. Although N. granulata is very tolerant to copper, acute exposure to this metal can disrupt the energy balance by affecting biochemical systems involved in carbohydrate metabolism.     

Recognition of co-existence pattern of salt marshes and mangroves for littoral forest restoration

Climate-change driven sea level rise causes a increase in salinity in coastal wetlands accelerating the alteration of the species composition. It triggers the gradual extinction of species, particularly the mangrove population which is intolerant of excessive salinity. Thus despite being crucial to a wide range of ecosystem services, mangroves have been identified as a vulnerable coastal biome. Hence restoration strategy of mangroves is undergoing rigorous research and experiments in literature at an interdisciplinary level. From a data-driven perspective, analysis of mangrove occurrence data could be the key to comprehend and predict mangrove behavior along different environmental parameters, and it could be important in formulating management strategy for mangrove rehabilitation and restoration. As salt marshes are the natural salt-accumulating halophytes, mitigating excessive salinity could be achieved by incorporating salt-marshes in mangrove restoration activities. This study intends to find a novel restoration strategy by assessing the frequent co-existence status of salt marshes, with the mangroves, and mangrove associates in different zones of degraded mangrove patches for species-rich plantation. To achieve this, we primarily design a novel methodological framework for the practice of knowledge discovery concerning the coexistence pattern of salt marshes, mangroves, and mangrove associates along with environmental parameters using a data mining paradigm of association rule mining. The proposed approach has the capability to uncover underlying facts and forecast likely facts that could automate the study in the field of ecological research to comprehend the occurrence of inter-species relationships. Our findings are based on published data gathered on the Sundarban Mangrove Forest, one of the world’s most important littoral forests. The existing literature reinforces the findings that include all the sets of frequently co-occurring mangroves, their associates, and salt marshes along the salinity gradient of coastal Sundarbans. A detailed understanding of the occurrence patterns of all these, along with the environmental variables, would be able to promote decision-making strategy. This framework is effective for both academia and stakeholders, especially the foresters/ conservation planners, to regulate the spread of salt marshes and the restoration of mangroves as well.     

Influence of abiotic factors on spider and ground beetle communities in different salt-marsh systems

Salt marshes are interesting and endangered ecosystems in West-Europe. Nevertheless, their arthropod fauna remains largely unknown and the factors determining assemblages at micro-habitat scale are poorly understood. Few data are also available about the effects of management measures in salt marshes and how to monitor them. The aim of the present study is to determine the major factors structuring two dominant communities of arthropods, spider and ground beetles, in natural, managed (cutting and sheep grazing) and invaded (by the grass Elymus athericus) salt marshes. The two taxa were studied during 2002 and 2003 in different salt marshes of the Mont Saint-Michel Bay (NW France) by pitfall traps and hand-collecting. A total of 12 350 spiders (57 species) and 16 355 ground beetles (34 species) were caught during the study and analysed with respect to effects of the salinity gradient and of habitat structure characteristics. Spiders and ground beetles reacted differently to environmental factors in salt marshes. Spiders could more easily cope with salinity and their presence/absence was less related to the salinity than that of ground beetles. For ground beetles, there were few other community-structuring environmental factors and these were only related to the edaphic environment: species restricted to open habitats, significant effects of moisture content and salinity revealed by CCA. Because they are likely to bring complementary information on abiotic factors, we finally suggest using both spiders and ground beetles for monitoring the effects of management practices in salt marshes.     

Responses of Salt Marsh Ecosystems to Mosquito Control Management Practices along the Atlantic Coast (U.S.A.)

Open marsh water management (OMWM) of salt marshes modifies grid‐ditched marshes by creating permanent ponds and radial ditches in the high marsh that reduce mosquito production and enhance fish predation on mosquitoes. It is preferable to using pesticides to control salt marsh mosquito production and is commonly presented as a restoration or habitat enhancement tool for grid‐ditched salt marshes. Monitoring of nekton, vegetation, groundwater level, soil salinity, and bird communities before and after OMWM at 11 (six treatment and five reference sites) Atlantic Coast (U.S.A.) salt marshes revealed high variability within and among differing OMWM techniques (ditch‐plugging, reengineering of sill ditches, and the creation of ponds and radial ditches). At three marshes, the dominant nekton shifted from fish (primarily Fundulidae species) to shrimp (Palaemonidae species) after manipulations and shrimp density increased at other treatment sites. Vegetation changed at only two sites, one with construction equipment impacts (not desired) and one with a decrease in woody vegetation along existing ditches (desired). One marsh had lower groundwater level and soil salinity, and bird use, although variable, was often unrelated to OMWM manipulations. The potential effects of OMWM manipulations on non‐target salt marsh resources need to be carefully considered by resource planners when managing marshes for mosquito control.     

A mixed strategy in the annual endemic Aster laurentianus (Asteraceae)--a stress-tolerant, yet opportunistic species

Several environmental factors influence the distribution of plants in coastal salt marshes. Substrate salinity is among the major factors preventing several species from establishing near the water line. However, interspecific competition for light and nutrients is often significant in determining the upper limit of plants along the salt marsh gradient. In this study, we tested the effects of substrate salinity and light and nutrient availability on the performance of the annual Aster laurentianus (Asteraceae), an endangered species of eastern Canadian salt marshes. This species is typically found in a narrow band along the shores of shallow lagoons, cornered between the high water line and the dense, herbaceous community of the upper marsh. Low light availability was the most significant factor limiting plant performance. Salinity had little effect on A. laurentianus as, unexpectedly, did nutrient availability. Yet plants were able to absorb nutrients when these were made more available. Luxury consumption, the uptake of excess nutrients, may make sense for this annual plant because the habitat in which it grows is subject to frequent disturbances (e.g., sand accretion and salinity pulses) that may kill canopy species and release suppressed A. laurentianus individuals. These results suggest that interspecific competition for light may play a significant role in restraining A. laurentianus from the upper part of salt marshes. Luxury consumption may help the species to opportunistically take advantage of release from taller species, particularly towards the upper edge of the salt marsh gradient.     

Are biotic factors significant in influencing the distribution of halophytes in saline habitats?

The influence of biotic factors on the distribution and establishment of halophytes is being considered in this review. Physicochemical factors, such as salinity and flooding, often are considered to be the determining factors controlling the establishment and zonational patterns of species in salt marsh and salt desert environments. Sharp boundaries commonly are found between halophyte communities even though there is a gradual change in the physicochemical environment, which indicates that biotic interactions may play a significant role in deterining the distribution pattern of species and the composition of zonal communities. Competition is hypothesized to play a key role in determining both the upper and lower limits of species distribution along a salinity gradient. Field and laboratory experiments indicate that the upper limits of distribution of halophytes into less saline or nonsaline habitats is often determined by competition. There appears to be a reciprocal relationship between the level of salt tolerance of species and their ability to compete with glycophytes in less saline habitats. Halophytes are not competitive in nonsaline habitats, but their competitive ability increases sharply in saline habitats. Allelopathic effects have been reported in salt desert habitats, but have not been reported along salinity gradients in salt marshes. Some species of halophytes that are salt accumulators have the ability to change soil chemistry. Chemical inhibition of intolerant species occurs when high concentrations of sodium are concentrated in the surface soils of salt desert plant communities that are dominated by salt-accumulating species. Establishment of less salt-tolerant species is inhibited in the vicinity of these salt-accumulating species. Herbivory is reported to cause both an increase and a decrease in plant diversity in salt marsh habitats. Heavy grazing is reported to eliminate sensitive species and produce a dense cover of graminoids in high marsh coastal habitats. However, in other marshes, grazing produced bare patches that allowed annuals and other low marsh species to invade upper marsh zonal communities. A retrogression in plant succession may occur in salt marshes and salt deserts because of heavy grazing. Intermediate levels of grazing by sheep, cattle, and horses could produce communities with the highest species richness and heterogeneity. Grazing by geese produced bare areas that had soils with higher salinity and lower soil moisture than vegetated areas, allowing only the more salt-tolerant species to persist. Removal of geese from areas by use of inclosures caused an increase in species richness in subarctic salt marshes. Invertebrate herbivores could also inhibit the survival of seeds and the ability of plants to establish in marshes. Parasites could play a significant role in determining the species composition of zonal communities, because uninfected rarer species are able to establish in the gaps produced by the death of parasitized species.     

Nitrogen further promotes a dominant salt marsh plant in an increasingly saline environment

Aims Human alterations of the environment are combining in unprecedented ways, making predictions of alterations to natural communities a difficult and pressing challenge. Estuarine systems have been subject to a high degree of modification, including increased nitrogen (N) inputs and altered salinity, factors important in shaping estuarine plant communities. As human populations increase and the climate changes, both N and salinity levels are likely to increase in these coastal marshes. Our objective was to evaluate the interactive effects of N and salinity on US West Coast salt marsh species; in particular, the performance of the dominant species Sarcocornia pacifica (pickleweed) alone and in mixed species assemblages. We expected increased salinity to favor S. pacifica but that N enrichment could help maintain greater species richness through use of N in salinity tolerance mechanisms.Methods We crossed treatments of N (added or not) and salinity (salt added or not) in a field experiment at a salt marsh in the San Francisco Estuary, California, USA, in each of three habitats: (i) monotypic pickleweed on the marsh plain, (ii) monotypic pickleweed along channels and (iii) mixed assemblages along channels. In a greenhouse experiment, we crossed treatments of N (added or not) and salinity (at three levels to simulate brackish to saline conditions) in (i) pots of pickleweed only and (ii) the same species mix as in the field.Important findings N addition doubled S. pacifica biomass and branching in both channel and marsh plain habitats regardless of salinity and greatly increased its dominance over Distichlis spicata and Jaumea carnosa in mixed assemblages along channels. In the greenhouse, S. pacifica biomass increased 6- to 10-fold with N addition over the range of salinities, while D. spicata and J. carnosa biomass increased with N addition only at lower salinity levels. Thus, while localized management could influence outcomes, expected overall increases in both N and salinity with human population growth and climate change are likely to enhance the production of S. pacifica in US West Coast marshes while reducing the diversity of mixed species assemblages. This decline in diversity may have implications for the resilience of marshes already subject to multiple stressors as the climate changes.     

Plathelminth abundance in North Sea salt marshes: environmental instability causes high diversity

Although supralittoral salt marshes are habitats of high environmental instability, the meiofauna is rich in species and abundance is high. The community structure of free-living Plathelminthes (Turbellaria) in these salt marshes is described. On an average, 104 individuals are found below an area of 10 cm². The average species density in ungrazed salt marshes is 11.3 below 10 cm² and 45.2 below 100 cm², indicating strong small-scale heterogenity. The faunal similarity between sediment and the corresponding above-ground vegetation is higher than between adjacent sample sites. Species prefer distinct ranges of salinity. In the lower part of the supralittoral salt marshes, the annual fluctuations of salinity are strongest and highly unpredictable. This region is richest in plathelminth species and abundance; diversity is highest, and the faunal composition of parallel samples is quite similar. In the upper part of the supralittoral salt marshes, the annual variability of salinity is lower, plathelminths are poor in species diversity and abundance. Parallel samples often have no species in common. Thus, those salt marsh regions with the most unstable environment are inhabited by the most diverse species assemblage. Compared to other littoral zones of the North Sea, however, plathelminth diversity in salt marshes is low. The observed plathelminth diversity pattern can apparently be explained by the dynamic equilibrium model (Huston, 1979).     

Anatomical adaptations and ionic homeostasis in aquatic halophyte Cyperus laevigatus L. Under high salinities

Salinity is extremely hazardous to agriculture worldwide and its expanding constantly. Soil of almost 100 countries facing salinity problem including Pakistan. Cyperus laevigatus also act as salinity indicator species is a naturally adapted halophyte dispersed in subtropical regions of world. Six populations of C. laevigatus were collected from different saline habitats to evaluate adaptations regarding anatomical and physiological characteristics. C. laevigatus is perfectly adapted to harsh environmental conditions like dry barren soils, saline lakes, hyper-saline wetlands and salt marshes. Ecological success of this species is due to plasticity in physiological and anatomical characteristics to adapt variable environmental conditions. C. laevigatus is a halophyte, exhibited increased biomass production in moderately saline habitat. Higher uptake of K⁺ occurs to compensate the uptake of Na⁺ ion contents, a striking feature of salt-tolerant and halophytic species. Accumulation of osmoprotectants like proline, free amino acids, soluble sugar and protein contribute significantly to osmotic adjustment. Stem thickness enhanced as salinity level of habitat increased to store water in parenchymatous tissues under physiological drought. Intensive sclerification in root cortex provide mechanical strength to plant as well as prevent the radial leakage of water. Well-developed aerenchyma, increased vascular bundle area, broader vessels, small and dense stomata are critical to cope with environmental hazards. Population of Jahlar lake showing maximum biomass production indicate that this species grows better in moderate salinities. Therefore, this species will prove very useful for revegetation of salt affected rangeland and prairies by direct growth of such halophytic ecotypes.     

Role of biotic interactions and physical factors in determining the distribution of marsh species along an estuarine salinity gradient

Understanding the mechanisms that shape plant distribution patterns is a major goal in ecology. We investigated the role of biotic interactions (competition and facilitation) and abiotic factors in creating horizontal plant zonation along salinity gradients in the Elbe estuary. We conducted reciprocal transplant experiments with four dominant species from salt and tidal freshwater marshes at two tidal elevations. Ten individuals of each species were transplanted as sods to the opposing marsh type and within their native marsh (two sites each). Transplants were placed at the centre of 9‐m² plots along a line parallel to the river bank. In order to disentangle abiotic and biotic influences, we set up plots with and without neighbouring vegetation, resulting in five replicates per site. Freshwater species (Bolboschoenus maritimus and Phragmites australis) transplanted to salt marshes performed poorly regardless of whether neighbouring vegetation was present or not, although 50–70% of the transplants did survive. Growth of Phragmites transplants was impaired also by competition in freshwater marshes. Salt marsh species (Spartina anglica and Puccinellia maritima) had extremely low biomass when transplanted to freshwater marshes and 80–100% died in the presence of neighbours. Without neighbours, biomass of salt marsh species in freshwater marshes was similar to or higher than that in salt marshes. Our results indicate that salt marsh species are precluded from freshwater marshes by competition, whereas freshwater species are excluded from salt marshes by physical stress. Thus, our study provides the first experimental evidence from a European estuary for the general theory that species boundaries along environmental gradients are determined by physical factors towards the harsh end and by competitive ability towards the benign end of the gradient. We generally found no significant impact of competition in salt marshes, indicating a shift in the importance of competition along the estuarine gradient.     

Neuroendocrine regulation of carbonic anhydrase expression in the gills of the euryhaline green crab, Carcinus maenas

The relationship between branchial carbonic anhydrase (CA) activity, CA gene expression and salinity, and potential mechanisms of regulation, was investigated in the euryhaline green crab, Carcinus maenas, acclimated to 33 ppt and transferred to 10 ppt, and the stenohaline rock crab, Cancer irroratus, acclimated to 32 ppt and transferred to 18 ppt. CA activity in green crabs acclimated to high and low salinity was a function of CA mRNA expression, with low salinity exposure resulting in an increase in both CA expression and activity. Eyestalk ablation (ESA) in green crabs acclimated to high salinity resulted in an increase in CA expression in the posterior, ion-transporting gills, in the absence of the low salinity stimulus. There were no changes in CA activity or expression in the anterior, respiratory gills. ESA also potentiated low salinity-stimulated CA induction, again, only in posterior gills. There were no changes in CA activity in any gills of Cancer irroratus, in response to either ESA or low salinity. These results suggest that CA expression in euryhaline, osmoregulating species, is under inhibitory regulation by a putative repressor found in the eyestalk, and that this mechanism is absent in stenohaline, osmoconforming species. CA expression is maintained at low, baseline levels in crabs acclimated to high salinity by the presence and action of this compound. The effects of the repressor appear to be reduced upon exposure to low salinity, allowing CA induction to occur.     

Salinity-mediated carbonic anhydrase induction in the gills of the euryhaline green crab, Carcinus maenas

The euryhaline green crab, Carcinus maenas, is a relatively strong osmotic and ionic regulator, being able to maintain its hemolymph osmolality as much as 300 mOsm higher than that in the medium when the crab is acclimated to low salinity. It makes the transition from osmoconformity to osmoregulation at a critical salinity of 26 ppt, and new acclimated concentrations of hemolymph osmotic and ionic constituents are reached within 12 h after transfer to low salinity. One of the central features of this transition is an 8-fold induction of the enzyme carbonic anhydrase (CA) in the gills. This induction occurs primarily in the cytoplasmic pool of CA in the posterior, ion-transporting gills, although the membrane-associated fraction of CA also shows some induction in response to low salinity. Inhibition of branchial CA activity with acetazolamide (Az) has no effect in crabs acclimated to 32 ppt but causes a depression in hemolymph osmotic and ionic concentrations in crabs acclimated to 10 ppt. The salinity-sensitive nature of the cytoplasmic CA pool and the sensitivity of hemolymph osmotic/ionic regulation to Az confirm the enzyme's role in ion transport and regulation in this species. CA induction is a result of gene activation, as evidenced by an increase in CA mRNA at 24 h after transfer to low salinity and an increase in protein-specific CA activity immediately following at 48 h post-transfer. CA gene expression appears to be under inhibitory control by an as-yet unidentified repressor substance found in the major endocrine complex of the crab, the eyestalk.