Effects of various salinities on Na(+)-K(+)-ATPase, Hsp70 and Hsp90 expression profiles in juvenile mitten crabs, Eriocheir sinensis

Eriocheir sinensis is a euryhaline crab migrating from sea to freshwater habitats during the juvenile stage. We used quantitative real-time polymerase chain reaction (qRT-PCR) to investigate the gene expression profile of Na(+)-K(+)-ATPase, Hsp70 (heat shock protein 70) and Hsp90 in megalopa exposed to salinities of 0, 2, 5, 10, and 15 parts per thousand. Both low and high salinities markedly stimulated expression of Na(+)-K(+)-ATPase, Hsp70 and Hsp90 genes of Chinese mitten crab megalopa; salinity had different effects on Na(+)-K(+)-ATPase, Hsp70 and Hsp90 levels depending on the duration of salinity stress, implying that Na(+)-K(+)-ATPase, Hsp70 and Hsp90 may play an important role in salinity tolerance in this crab species.     

Salinity tolerance of adult and juvenile red king crabs Paralithodes camtschatica

• 1.1. Juvenile king crabs were more tolerant of reduced salinities than adult crab; juvenile crab were better volume regulators at reduced salinities than adult crab.
• 2.2. Adult female king crab hemolymph was hyperosmotic to full seawater (30 ppt) and isosmotic to dilute seawater. Juvenile king crab (2 years old) were hypoosmotic at the same concentrations.
• 3.3. Lower osmotic concentration of juvenile hemolymph is at least partially due to lower sodium concentration.
• 4.4. Juvenile king crab can tolerate some dilution and survive for short periods in the reduced salinity of the lower intertidal zone.

     

The Na+, K+, 2Cl- cotransporter of estuarine pufferfishes (Sphoeroides testudineus and S. greeleyi) in hypo- and hyper-regulation of plasma osmolality

The pufferfishes Sphoeroides testudineus and Sphoeroides greeleyi are estuarine species that osmoregulate efficiently, but S. testudineus tolerates seawater dilution to a much higher degree than S. greeleyi. This study aimed at testing whether NKCC is involved with their differential tolerance of seawater dilution, through the analysis of in vivo furosemide (NKCC inhibitor) injection both on hypo-regulation (in 35 per thousand salinity) and hyper-regulation (in 5 per thousand salinity). After exposure for 6 h or 5 days to both salinities, blood samples were obtained for determination of plasma osmolality, chloride, sodium and hematocrit, and muscle samples for determination of water content. Furosemide injection led to increased plasma osmolality and sodium in 35 per thousand and decreased osmolality and chloride in 5 per thousand, when compared to saline-injected controls. Furosemide injection led to hematocrit reduction in both salinities, and muscle water content increase in 5 per thousand and decrease in 35 per thousand in S. testudineus. The results are compatible with NKCC working in branchial NaCl secretion in 35 per thousand, in both species, and a higher role in cell volume regulation in blood and muscle cells of S. testudineus, in both salinities, which could partially explain the stronger capacity of S. testudineus to tolerate seawater dilution during low tide.     

Physiological response in the European flounder (Platichthys flesus) to variable salinity and oxygen conditions

Physiological mechanisms involved in acclimation to variable salinity and oxygen levels and their interaction were studied in European flounder. The fish were acclimated for 2 weeks to freshwater (1 per thousand salinity), brackish water (11 per thousand) or full strength seawater (35 per thousand) under normoxic conditions (water Po(2) = 158 mmHg) and then subjected to 48 h of continued normoxia or hypoxia at a level (Po(2) = 54 mmHg) close to but above the critical Po(2). Plasma osmolality, [Na(+)] and [Cl(-)] increased with increasing salinity, but the rises were limited, reflecting an effective extracellular osmoregulation. Muscle water content was the same at all three salinities, indicating complete cell volume regulation. Gill Na(+)/K(+)-ATPase activity did not change with salinity, but hypoxia caused a 25% decrease in branchial Na(+)/K(+)-ATPase activity at all three salinities. Furthermore, hypoxia induced a significant decrease in mRNA levels of the Na(+)/K(+)-ATPase alpha1-subunit, signifying a reduced expression of the transporter gene. The reduced ATPase activity did not influence extracellular ionic concentrations. Blood [Hb] was stable with salinity, and it was not increased by hypoxia. Instead, hypoxia decreased the erythrocytic nucleoside triphosphate content, a common mechanism for increasing blood O(2) affinity. It is concluded that moderate hypoxia induced an energy saving decrease in branchial Na(+)/K(+)-ATPase activity, which did not compromise extracellular osmoregulation.     

Oxygen consumption and osmoregulatory capacity in Neomysis integer reduce competition for resources among mysid shrimp in a temperate estuary

Results of field surveys and laboratory measurements of oxygen consumption and body fluid osmolality at different salinities in the mysids Neomysis integer, Mesopodopsis slabberi, and Rhopalophthalmus mediterraneus from the Guadalquivir estuary (southwest Spain) were used to test the hypothesis that osmotic stress (oxygen consumption vs. isosmotic points) was lowest at salinities that field distributions suggest are optimal. The three species showed overlapping spatial distributions within the estuary but clear segregation along the salinity gradient: N. integer, M. slabberi, and R. mediterraneus displayed maximal densities at lower, intermediate, and higher salinities, respectively. Adults of N. integer were extremely efficient hyperregulators (isosmotic point 30 per thousand) over the full salinity range tested (3 per thousand-32 per thousand), and their oxygen consumption rates were independent of salinity; adults of M. slabberi were strong hyper- and hyporegulators at salinities between 7 per thousand and 29 per thousand (isosmotic point, 21 per thousand) and showed higher oxygen consumptions at the lowest salinity (6 per thousand); adults of R. mediterraneus hyperregulated at salinities between 19 per thousand and seawater (isosmotic point, 36 per thousand), with the lowest oxygen consumption at salinity around their isosmotic point (35 per thousand). Thus, the osmoregulation capabilities of M. slabberi and R. mediterraneus seem to determine the salinity ranges in which most of their adults live, but this is not so for adults of N. integer. Moreover, maximal field densities of M. slabberi (males and females) and R. mediterraneus (males) occur at the same salinities as the lowest oxygen consumption. In contrast, field distribution of N. integer was clearly biased toward the lower end of the salinity ranges within which it osmoregulated. We hypothesize that the greater euryhalinity of N. integer makes it possible for this species to avoid competition with R. mediterraneus by inhabiting the more stressful oligohaline zone.     

Effects of long-term exposure to different salinities on the location and activity of Na+-K+-ATPase in the gills of juvenile mitten crab, Eriocheir sinensis

The euryhalinity of mitten crab, Eriocheir sinensis, is based on osmoregulation, and thus on the activity of Na(+)-K(+)-ATPase. We studied location and activity of this enzyme in gills of juvenile crabs exposed to 5 per thousand, 25 per thousand, and 40 per thousand salinity. The posterior gills showed always a high number of immunopositive cells (IPC), staining with fluorescent antibody against Na(+)-K(+)-ATPase, covering at 5 per thousand the entire lamellae. At 25 per thousand, they showed fewer IPC which occurred only at the bases of the lamellae. Enzyme activity was consistently higher in posterior than in anterior gills. Low salinity stimulated the activity only in posterior gills. Both histochemical and enzymatic results are consistent with previous ultrastructural observations showing that the epithelial cells of the posterior, but not the anterior gills exhibit typical traits of ionocytes. While an increase in Na(+)-K(+)-ATPase activity at a reduced salinity is consistent with a strong hyper-osmoregulatory capacity in juvenile crabs, a low activity at an enhanced salinity suggests a physiological response, directed towards a reduction of Na(+) uptake. The activity increase of ion-transporting enzymes is directly related to spatial changes in their distribution along the osmoregulatory tissue, i.e. an enhanced number of IPC scattered along the entire lamellae. In juveniles, this allows for successful development and growth at reduced salinities.     

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.     

Ontogeny of osmoregulation and salinity tolerance in a mangrove crab, Sesarma curacaoense (Decapoda: Grapsidae)

The grapsid crab Sesarma curacaoense is believed to represent the closest saltwater relative to the ancestor which gave rise to an adaptive radiation of endemic freshwater and terrestrial species on the island of Jamaica. Living in mangrove swamps with variable salinity conditions and showing semiterrestrial behaviour, S. curacaoense exhibits ecological adaptations to non-marine conditions. In laboratory experiments, we studied the salinity tolerance during development from hatching to the end of the first juvenile stage. Successful development through metamorphosis occurred in the full salinity range tested (15-32 per thousand), although mortality was significantly enhanced and development delayed at 15 per thousand. In another series of experiments, we studied the ontogeny of the capability for osmoregulation, which is considered as the physiological basis of osmotic stress tolerance. Our results show that S. curacaoense is from hatching a fairly strong hyperosmoregulator in dilute media. This capability increased gradually from hatching throughout the larval and juvenile development. In seawater (32 per thousand) and at an enhanced salt concentration (44 per thousand), the zoeal stages remained hyperosmoconformers. The capability for hypoosmoregulation in concentrated media appeared first in the megalopa stage and increased thereafter. Adult crabs were observed to be strong hyper-hypo-osmoregulators in a salinity range from at least 1 per thousand to 44 per thousand. The unusually early appearance of strong regulatory capabilities, particularly in dilute media, is interpreted as a physiological preadaptation that should have facilitated the evolutionary process of adaptive radiation in non-marine environments on Jamaica.     

Behavioral influences on the physiological responses of Cancer gracilis, the graceful crab, during hyposaline exposure

The relationship between the behavioral and physiological responses to hyposaline exposure was investigated in Cancer gracilis, the graceful crab. The status of C. gracilis as an osmoconformer was confirmed. Survival decreased with salinity: the LT(50) in 50% seawater (a practical salinity of 16, or 16 per thousand) was 31.5 +/- 22.7 h and in 25% seawater (a salinity of 8) was 8.0 +/- 0.7 h. When exposed to a salinity gradient, most crabs moved towards the highest salinity. However, in the salinity range of 55% to 65% seawater, they became quiescent. This "closure response" was also evident at low salinities: the mouthparts were tightly closed and animals remained motionless for 2 to 2.5 h. During closure, crabs were able to maintain the salinity of water within the branchial chambers at a level that was about 30% higher than that of the surrounding medium. The closure response was closely linked to a short-term decrease in oxygen uptake. During closure, oxygen within the branchial chamber was rapidly depleted, with oxygen uptake returning to pretreatment levels upon the resumption of activity. In addition to the short-term decrease in oxygen uptake, there was a longer-term bradycardia, which may serve to further reduce diffusive ion loss across the gills. By exhibiting a closure response during acute hyposaline exposure and an avoidance reaction during prolonged or severe hyposaline exposure, C. gracilis is able to use behavior to exploit areas prone to frequent episodes of low salinity.     

Branchial osmoregulatory response to salinity in the gilthead sea bream, Sparus auratus

The branchial osmoregulatory response of gilthead sea bream (Sparus auratus L.) to short-term (2-192 hr) and long-term (2 weeks) exposure to different environmental salinities (5 per thousand, 15 per thousand, 25 per thousand, 38 per thousand and 60 per thousand) was investigated. A "U-shaped" relationship was observed between environmental salinity and gill Na+,K+ -ATPase activity in both long- and short-term exposure to altered salinity, with the increase in activity occurring between 24 and 96 hr after the onset of exposure. Plasma osmolality and plasma ions (sodium, chloride, calcium and potassium) showed a tendency to increase in parallel with salinity. These variables only differed significantly (P<0.05) in fish adapted to 60 per thousand salinity with respect to fish adapted to full-strength sea-water (SW). Plasma glucose remained unchanged whereas plasma lactate was elevated at 5 per thousand and 60 per thousand. Muscle water content (MWC) was significantly lower in fish adapted to 60 per thousand. Chloride cells (CC) were only present on the surface of the gill filaments and absent from the secondary lamellae. CC distribution was not altered by external salinity. However, the number and size of CC were significantly increased at salinity extremes (5 per thousand and 60 per thousand), whereas fish exposed to intermediate salinities (15 per thousand and 25 per thousand) had fewer and smaller cells. Furthermore, the CC of fish exposed to diluted SW became rounder whereas they were more elongated in fish in full-strength and hypersaline SW. This is consistent with previous reports indicating the existence of two CC types in euryhaline fish. At likely environmental salinities, gilthead sea bream show minor changes in plasma variables and the effective regulation of gill Na+,K+ -ATPase. However, at very low salinities both haemodilution and up-regulation of gill Na+,K+ -ATPase predict a poor adaptation most likely related to deficiency or absence of specific components of the CC important for ion xuptake.     

Effect of salinity changes on the bacterial diversity, photosynthesis and oxygen consumption of cyanobacterial mats from an intertidal flat of the Arabian Gulf

The effects of salinity fluctuation on bacterial diversity, rates of gross photosynthesis (GP) and oxygen consumption in the light (OCL) and in the dark (OCD) were investigated in three submerged cyanobacterial mats from a transect on an intertidal flat. The transect ran 1 km inland from the low water mark along an increasingly extreme habitat with respect to salinity. The response of GP, OCL and OCD in each sample to various salinities (65 per thousand, 100 per thousand, 150 per thousand and 200 per thousand) were compared. The obtained sequences and the number of unique operational taxonomic units showed clear differences in the mats' bacterial composition. While cyanobacteria decreased from the lower to the upper tidal mat, other bacterial groups such as Chloroflexus and Cytophaga/Flavobacteria/Bacteriodetes showed an opposite pattern with the highest dominance in the middle and upper tidal mats respectively. Gross photosynthesis and OCL at the ambient salinities of the mats decreased from the lower to the upper tidal zone. All mats, regardless of their tidal location, exhibited a decrease in areal GP, OCL and OCD rates at salinities > 100 per thousand. The extent of inhibition of these processes at higher salinities suggests an increase in salt adaptation of the mats microorganisms with distance from the low water line. We conclude that the resilience of microbial mats towards different salinity regimes on intertidal flats is accompanied by adjustment of the diversity and function of their microbial communities.     

Possible role of carbonic anhydrase, V-H(+)-ATPase, and Cl(-)/HCO3- exchanger in electrogenic ion transport across the gills of the euryhaline crab Chasmagnathus granulatus

We studied the participation of carbonic anhydrase (CA), V-H(+)-ATPase, and Cl(-)/HCO3- exchanger in electrogenic ion absorption through the gills of Chasmagnathus granulatus. CA activity was measured in anterior gills and posterior gills after acclimation to 2 per thousand, 10 per thousand, 30 per thousand (about seawater), and 45 per thousand salinity. The highest CA specific activity was detected in the microsomal fraction in anterior gills, and in the cytosolic fraction, in posterior ones. Both fractions were strongly induced by decreasing salinity only in posterior gills. Perfusion of posterior gills from crabs acclimated to either 2 per thousand or 10 per thousand with acetazolamide inhibited CA activity almost completely. In posterior gills from crabs acclimated to 2 per thousand and perfused with 20 per thousand saline (iso-osmotic for these crabs), acetazolamide reduced transepithelial potential difference (V(te)) by 47%, further addition of ouabain enhanced the effect to 88%. Acetazolamide had no effect in the same gills perfused with 30 per thousand saline (iso-osmotic for seawater acclimated crabs). Bafilomycin A1 and SITS (inhibitors of V-H(+)-ATPase and Cl(-)/HCO3-) reduced V(te) by 15-16% in gills perfused with normal 20 per thousand saline, and by 77% and 45%, respectively when they were applied in Na-free 20 per thousand saline, suggesting the participation of those transporters and cytosolic CA in electrogenic ion absorption.     

Distinct patterns of water and osmolyte control between intertidal (Bunodosoma caissarum) and subtidal (Anemonia sargassensis) sea anemones

Anemones are frequently found in rocky intertidal coasts. As they have highly permeable body surfaces, exposure to the air or to salinity variations inside tidal pools can represent intense osmotic and ionic challenges. The intertidal Bunodosoma caissarum has been compared with the subtidal Anemonia sargassensis concerning their response to air exposure or salinity changes. B. caissarum maintains tissue hydration through mucus production and dome-shape formation when challenged with air exposure or extreme salinities (fresh water or hypersaline seawater, 45 psu) for 1-2h. Upon exposure to mild osmotic shocks for 6h (hyposmotic: 25 psu, or hyperosmotic: 37 psu), B. caissarum was able to maintain its coelenteron fluid (CF) osmolality stable, but only in 25 psu. A. sargassensis CF osmolality followed the external medium in both salinities. Isolated cells of the pedal disc of B. caissarum showed full capacity for calcium-dependent regulatory volume decrease (RVD) upon 20% hyposmotic shock, at least partially involving the release of KCl via K(+)-Cl(-) cotransport, and also of organic osmolytes. Aquaporins (HgCl(2)-inhibited) likely participate in this process. Cells of A. sargassensis showed partial RVD, after 20 min. Cells from both species were not capable of regulatory volume increase upon hyperosmotic shock (20%). Whole organism and cellular mechanisms allow B. caissarum to live in the challenging intertidal habitat, frequently facing air exposure and seawater dilution.     

Osmo and ionic regulation of black tiger prawn (Penaeus monodon Fabricius 1798) juveniles exposed to K(+) deficient inland saline water at different salinities

An 11-day trial was conducted to investigate the osmoregulatory capacity (OC) and regulation of K(+), Na(+), Ca(2+) and Mg(2+) of Penaeus monodon juveniles when exposed to K(+) deficient inland saline water (ISW) of four different salinities (5, 15, 25 and 35 ppt). The survival of juveniles showed a positive linear relationship (R(2) ranging from 0.72 to 0.98) with salinity. At the end of the trial, juveniles were able to survive only in 5 ppt of ISW and showed no changes in OC when transferred from ocean water (OW) to ISW. Further, the OC of juveniles in 5 ppt of ISW was significantly different (P<0.05) from the OC of juveniles exposed to 15, 25 and 35 ppt and exhibited strong serum K(+), Na(+), Ca(2+) and Mg(2+) regulation monitored over 16 h. In contrast, at 35 ppt, significant decrease (P<0.05) in serum K(+) and Mg(2+) concentrations and accumulation of serum Na(+) concentration occurred after 16 h of exposure to ISW. At higher salinity, an increase in serum Na(+) concentration leads to an increase in the serum osmolality of the juveniles, which in turn causes decrease in the OC of the juveniles. The results of this study suggest that K(+) deficiency in ISW has a negative effect on survival, OC and the ability of P. monodon juveniles to regulate serum Na(+), K(+), Ca(2+) and Mg(2+) concentrations. These effects are compounded as salinity increases.     

Effect of salinity on oxygen consumption in fishes: a review

The effect of salinity on resting oxygen uptake was measured in the perch Perca fluviatilis and available information on oxygen uptake in teleost species at a variety of salinities was reviewed. Trans‐epithelial ion transport against a concentration gradient requires energy and exposure to salinities osmotically different from the body fluids therefore imposes an energetic demand that is expected to be lowest in brackish water compared to fresh and sea water. Across species, there is no clear trend between oxygen uptake and salinity, and estimates of cost of osmotic and ionic regulation vary from a few per cent to >30% of standard metabolism.     

Drinking and osmoregulation in the mangrove crab Ucides cordatus following exposure to benzene

In benzene-exposed Ucides cordatus acclimated for 96 h to 9 and 34 per thousand SW, haemolymph, urine and gastric juice are isosmotic with each other, but differ significantly in osmolality from external media. In both salinities, under benzene action, urine K+ excretion and calcium absorption are increased significantly, whereas Na+ absorption and Mg2+ excretion show U/B ratios similar to control values. In 9 per thousand SW, some ionic exchanges via benzene-exposed gills are possibly hastened. Benzene exposure decreases significantly branchial chamber water osmolality, [Na+] and [K+], whereas [Ca2+] and [Mg2+] are unaffected. However, faster medium exchange presumably occurs in 34 per thousand SW, both crab groups show branchial chamber water osmotic and ionic concentrations similar to surrounding medium. Benzene exposure unaffected gastric juice composition. In both media, [Ca2+] and [Mg2] accumulate several times higher than surrounding media, and [Na+] and [K+] are significantly hypo-ionic to haemolymph. Na+ and K+ G/H ratios are lower in crabs acclimated to 34 per thousand SW than in crabs acclimated to 9 per thousand SW. Drinking rates are enhanced by benzene exposure and are higher at 34 per thousand SW than in seawater isosmotic with the haemolymph (26 per thousand SW). Benzene exposure affects significantly osmoregulatory capability, slowing haemolymph dilution after transfer to clean 9 per thousand SW. Lower haemolymph dilution rate accounts for higher osmolality, but 48 h after transfer there is no recovery like in control crabs. Haemolymph transfusion experiments suggest an interaction among effects of benzene and hormonal factors, possibly on water influx.     

Water Balance in the Land Crab, Gecarcinus lateralis, During the Intermolt Cycle

Gecarcinus lateralis can take moisture from a clamp substratumin amounts adequate for the needs of the entire intermolt cycle.It can also rehydrate in this way, even after severe dehydration. This crab is able to survive for many months when free waterof a wide range of salinities (0.30; = parts per thousand)is made available in a shallow dish. The crab dies within sevenweeks when the salinity of this water is 35. During proecdysisthe pericardial sacs of eyestalkless crabs become most swollenwhen the salinity of the available water is 15 or 23, and survivalduring and after ecdysis is greatest with water of 15. A crab in proecdysis shows no increase in the rate at whichwater enters following dehydration. Yet large amounts of waterare retained, particularly at the intermediate salinities. Maximalswelling of the pericardial sacs just prior to ecdysis is essentiallyequivalent in crabs with eyestalks, in eyestalkless crabs, andin eyestalkless crabs that have received an implant of centralnervous tissue. Hence, we conclude that a hormone causing theretention of water exists, but not in the eyestalks, in thebrain, or in the thoracic ganglionic mass. At ecdysis eyestalkless crabs show large increases in the dimensionsof the carapace, while crabs with eyestalks and eyestalklesscrabs that have received an implant of certain central nervoustissues show much less increase and may even show a decrease.Thus, we conclude that a hormone causing a release of waterat ecdysis is produced in the central nervous system. The advantages to the crab of a dual hormonal control of itswater balance are discussed.     

Salt and water balance in the sipunculan phascolopsis gouldi: is any animal a “simple osmometer”?

• 1.1. Phascolopsis gouldi, the commonly studied sipunculan of the Woods Hole area, Massachusetts, can tolerate a salinity range from about 45% seawater to at least 100% SW, with literature records extending to about 160% SW. There was no survival at 40% SW.
• 2.2. Over this salinity range, P. gouldi is an osmotic and ionic conformer.
• 3.3. The osmotic and sodium concentrations of the coelomic fluid are the same as that of the medium; the chloride concentration is about 9% less than that of the medium; the potassium concentration is about 30% higher.
• 4.4. Analysis of water content regulation by two different approaches shows that P. gouldi does have a limited ability for volume regulation, restricted to salinities higher than 58% SW.
• 5.5. P. gouldi is not a “simple osmometer”.

     

Effect of salinity on osmoregulatory patch epithelia in gills of the blue crab Callinectes sapidus

In euryhaline crabs, ion-transporting cells are clustered into osmoregulatory patches on the lamellae of the posterior gills. To examine changes in the branchial osmoregulatory patch in the blue crab Callinectes sapidus in response to change in salinity and to correlate these changes with other osmoregulatory responses, crabs were acclimated to a range of salinities between 10 and 35 ppt. When crabs that had been acclimated to 35 ppt were subsequently transferred to 10 ppt, both the size of the osmoregulatory patch on individual gill lamellae and the specific activity of Na+, K+-ATPase in whole-gill homogenates increased only after the first 24 h of exposure to dilute seawater. Enzyme activity and size of patch area increased gradually and reached their maxima (increasing by 200% and 60%, respectively) 6 days following transfer to 10 ppt seawater and then remained at these levels. Patch size at acclimation varied inversely with the salinity for seawater dilutions below 26 ppt (the isosmotic point of the crab), although it did not vary in salinities at or above 26 ppt. Thus, the size of the patch clearly is modulated with acclimation salinity, but it increases only in those salinities in which the crab hyperosmoregulates. An increase in the total RNA/DNA ratio in gill homogenates, the lack of mitotic figures in the lamellae, and the lack of incorporation of bromodeoxyuridine into nuclei of lamellar epithelial cells during acclimation to dilute seawater were interpreted as evidence that no cell proliferation had occurred and that increases in the size of the osmoregulatory patch occurred through differentiation of existing gas exchange cells or of undifferentiated epithelial cells into ion-transporting cells.     

Physiological and behavioral responses of the mud snails Hydrobia glyca and Hydrobia ulvae to extreme water temperatures and salinities: implications for their spatial distribution within a system of temperate lagoons

Physiological responses (oxygen consumption) and behavioral responses (feeding and activity) of the mud snails Hydrobia ulvae and Hydrobia glyca at different salinities (20 per thousand-80 per thousand) and temperatures (20 degrees and 30 degrees C) were studied. After 24 h under experimental conditions, both Hydrobia species already showed maximal activities (>90%) for a wide salinity range (30 per thousand-70 per thousand), with significant differences in activity between species only outside the usual salinity range of the studied lagoon. In contrast, egestion rates of H. glyca were significantly higher at the lowest salinities tested (30 per thousand and 40 per thousand) irrespective of water temperature, whereas egestion rates of H. ulvae were always significantly higher (57% on average) at 20 degrees C than at 30 degrees C and at the usual salinities found in the field (40 per thousand and 50 per thousand). Both species showed an oxyregulatory response to dissolved oxygen concentrations ranging from saturation to 1.5 mg O(2) L(-1), although specific oxygen consumption rates were significantly higher at 30 degrees C than at 20 degrees C (Q(10)=1.47+/-0.08 for H. ulvae and Q(10)=12.1+/-0.06 for H. glyca) and at the lowest salinities (30 per thousand-50 per thousand for H. ulvae and 30 per thousand-40 per thousand for H. glyca). On average, specific rates were higher for the smaller-sized H. glyca (1.64+/-0.03 microg O(2) mg(-1) ash-free dry weight [AFDW]) than for H. ulvae (1.35+/-0.03 microg O(2) mg(-1) AFDW). Despite the overlapping of their tolerances to high temperatures and salinities, the observed interspecies differences could play a certain role in the distribution of H. ulvae and H. glyca in the studied habitat. In particular, the decreasing feeding activity but increasing respiration of H. ulvae at 30 degrees C for salinities that usually occur in the studied lagoon could represent disadvantages to H. glyca during the warm period.