Conductive sodium entry in gill cells of the shore crab,Carcinus maenas

Isolated posterior gills of shore crabs, Carcinus maenas, collected from the Baltic Sea, were perfused and bathed with sea water and solutions of alkali chlorides. The preparation was used to measure fluxes of sodium from the external medium across the gills into the hemolymph and to determine transepithelial potential differences (PDs). Internally negative active transport PDs resulted from perfusion and bathing the gills symmetrically with the same medium (50% sea water). Passive (asymmetry) PDs following employment of 100% sea water as internal and 50% sea water as external medium were — in contrast to symmetry PDs — insensitive to cyanide and ouabain. This result indicates that the gill recognized the desired hyperosmotic state and responded by switching off the active transport component observed under symmetry conditions. Diffusional potential differences and fluxes of Na⁺ were inhibited by the externally applied diuretic amiloride. Gradients of pure alkali chlorides between medium and blood were accompanied by cation specific PDs. These PDs and their amiloride sensitivity were inversely related to the diameter of the unhydrated cation and allowed the calculation of the permeability sequence P: Li⁺>Na⁺>K⁺>Rb⁺>Cs⁺>Cl^-. The results obtained show that the permeability of the gills to cations greatly exceeds that to anions. In addition, these findings indicate that the initial amiloride-sensitive step in Na⁺ transport across the gill is not represented by an electroneutral sodium/proton exchange but by a conductive mechanism.     

Mechanisms of copper toxicity in the shore crab,Carcinus maenas

The influence of copper(II)chloride on selected key enzymes and metabolites in intermediary metabolism and the energy charge potential of the hyperregulating shore crab, Carcinus maenas (L.) were investigated. Crabs, collected in Odense Fjord, Denmark between September 1989 and May 1990, were exposed to 10 ppm copper(II)chloride, in 10 ppt salinity seawater at 15°C for up to 1 wk. Hexokinase activity was 77% lower in the posterior gills and 60% lower in the midgut gland than in control crabs. Phosphofructokinase activity was reduced in the midgut gland by 82% but was unaffected in the gills. Pyruvate kinase activity was undetectable in the gills of exposed crabs and reduced by 47% in the midgut gland. Citrate synthase activity was depressed by 30% in the posterior gills and unaffected in other tissues. Copper exposure had no apparent effect on tissue cytochrome c oxidase activity. Chela muscle was the only tissue in which copper exposure did not bring about reductions in the enzyme activities tested. As a result of 1 wk of copper exposure the average lactate level increased by 3.5-fold in haemolymph, 9.3-fold in posterior gills and 6.5-fold in midgut gland. The average glucose level was raised by a factor of 6 in the haemolymph, 3.4 in the posterior gills and 1.2 in the midgut gland. Energy charge potential was at a very high level (ca. 0.9) and was unaffected by copper exposure.     

Influence of cadmium accumulation and dietary status on fatty acid composition in two colour forms of shore crabs, Carcinus maenas

 The effects of cadmium exposure and dietary status on cadmium accumulation, fatty acid (FA) content and profiles were investigated in two colour forms of the shore crab Carcinus maenas. Groups of shore crabs were either starved or fed with blue mussels, Mytilus edulis, during a 40 d exposure period to 2 or 6 μM Cd²⁺ (as CdCl₂). Starved green individuals accumulated more cadmium in haemolymph and hepatopancreas than did red crabs and green crabs fed during the experiments. In the red colour form, no difference in cadmium accumulation was observed between starved and fed individuals. In both colour forms, hepatopancreas contained more FA than gills and muscle. The FAs often present in the largest amounts in the tissues were 16:0, 16:1ω7, 18:1ω7, 18:1ω9, 20:4ω6, 20:5ω3 and 22:6ω3. However, saturated (SAFA) and mono-unsaturated fatty acids (MUFAs) were dominant in hepatopancreas, whereas poly-unsaturated fatty acids (PUFAs) were dominant in gills and muscles. At the beginning of the experiment, the total FA content in the hepatopancreas was 111.6 mg g⁻¹ (dry weight) for red crabs and 78.4 mg g⁻¹ for green shore crabs. During the experiment, however, the FA content decreased in red crabs. This decrease was more pronounced for starved individuals than for fed individuals. Also, the decrease in FA content was more pronounced in crabs exposed to 6 μM cadmium compared to crabs exposed to 2 μM or crabs not exposed to cadmium. No change in FA content was observed in green shore crabs, irrespective of diet and cadmium exposure. For both colour forms, no change in FA content was observed for gills and muscle. In red crabs, a decrease was observed for all FAs in the hepatopancreas. This decrease, however, was more pronounced for SAFAs and MUFAs than for PUFAs, indicating that the metabolism of FAs during starvation and cadmium exposure is selective. The experiments indicate that green colour forms of shore crabs are more tolerant of natural stress such as starvation and anthropogenic stress, e.g. cadmium exposure, than are red colour forms of shore crabs. Received: 23 September 1999 / Accepted: 29 April 2000     

Arsenic accumulation in the shore crab Carcinus maenas: the influence of nutritional state,sex and exposure concentration

The accumulation of arsenate from seawater by the shore crab Carcinus maenas L. (collected from Odense Fjord, Denmark in 1991 and from Restronguet Creek, UK in 1991) was investigated in a series of laboratory experiments. A field study was also carried out to determine the effects of raised environmental arsenic concentrations on intra-organismal distribution and tissue concentrations. Studies on the influence of nutritional state and sex on accumulation of As(5) from seawater indicated that most of the arsenic taken up from seawater in laboratory experiments was retained in the gills and the midgut gland. Arsenic accumulation exhibited sex-dependent differences which were also evident in correlation analyses carried out between total lipid contents and total arsenic contents of midgut glands of individual crabs. Arsenic concentrations in the gonads of both sexes were strongly influenced by the nutritional state of the crabs. Elevated arsenic concentrations in seawater and food at an arsenic polluted site (Restronguet Creek) significantly influenced arsenic concentrations and distribution among the tissues of C. maenas. Arsenic concentrations and distribution patterns differed markedly from those crabs from an unpolluted site in Odense Fjord. The gills of the crabs from Restronguet Creek contained extremely high arsenic concentrations ranging from 179 to 483 g As g^-1 dry wt. These values were even higher than those measured in the gills of Odense crabs that had been exposed to 3 mgl^-1 As(5) for 2 wk in the laboratory. Arsenic concentrations in the exoskeleton of Odense Fjord crabs were 15 times lower than those measured in exoskeletons of Restronguet Creek crabs. Approximately 69% of the total body burden of arsenic was located in muscle tissue of crabs from Odense Fjord, whereas the major pool of arsenic (46%) in Restronguet Creek crabs was located in the exoskeleton.     

Ultrastructural changes in the gill epithelium of the crab Chasmagnathus granulatus (Decapoda: Grapsidae) in diluted and concentrated seawater

Chasmagnathus granulatus Dana, 1851 is an intertidal estuarine crab that experiences acute salinity changes ranging from <1‰ to full-strength seawater and even hypersaline waters in tide pools concentrated by evaporation. Ultrastructural changes induced by salinity in the posterior gills were examined in crabs collected from the Rio de la Plata estuary Argentina during March 1999. The posterior gills of C. granulatus are involved both in ion uptake and ion secretion depending on the acclimation medium. These organs are mostly lined with a thick tissue, which presents the characteristics of a typical salt-transporting epithelium. Electron microscopy analysis of gill tissue from crabs acclimated to dilute, full, and concentrated seawater (12‰, 34‰, and 45‰ salinity) showed significant development of basolateral membrane interdigitations, with numerous mitochondria and conspicuous apical membrane infoldings. Morphometrical analysis indicated that the subcuticular space delimited by the infolding of the apical membrane was significantly increased in the gills of high-salinity acclimated crabs. Septate junctions, which are thought to define the paracellular permeability, were significantly shorter in high-salinity acclimated crabs, suggesting a possible role of the paracellular pathway in salt secretion.     

Kinetics of osmoregulation in the crab Carcinus maenas

When shore crabs Carcinus maenas are transferred from 11 to 38 S at 11°C, new constant levels of hemolymph freezing points and of concentrations of Na, K, Ca, and Mg in the hemolymph are accomplished within 24h. From a decrease in serum protein and in serum free amino acids and an increase in the relative amounts of individual essential free amino acids in the body fluids of whole crabs, a participation of proteolytic activities and a transport of amino acids from hemolymph to cell is deduced; the stationary concentration of total intracellular free amino acids increases up to a nearly two-fold value, compared with the concentration in crabs remaining in diluted sea water. Also, the low molecular neutral sugars increase in whole crabs after high salinity stress, reaching values of more than two-fold initial concentrations within a period of 10 days. This increase is fully accounted for by a 6.7-fold increase in trehalose levels. The processes of increasing the concentrations of low molecular organic material seem to be slower than those of establishing new osmotic pressures in the hemolymph. The oxygen consumption decreases by 30 to 45% to new constant values within 8 to 12 h after the salinity change from 11 to 38, reflecting similar kinetics as the establishment of new osmotic pressures in the hemolymph.     

Sodium regulation in the shore crab Carcinus maenas as related to ambient salinity

The activity of Na–K-ATPase was measured in crude homogenates prepared from various organs (leg muscle, pincer muscle, heart, testes, digestive gland, hypodermis, gills 1–9) of shore crabs, Carcinus maenas L., acclimated to salinities ranging between 10 and 50 S (in steps of 10 S). In all salinities tested, Na–K-ATPase activity was highest in posterior gills 7–9 (10–12 mol P_i mg protein^-1 h^-1), followed by anterior gills 1–6 (ca. 2.5 mol P_i mg protein^-1 h^-1) and the other organs (in most cases far below 2mol P_i mg protein^-1 h^-1). In gills only, Na–K-ATPase activity was salinity-dependent, with the highest values in the lowest salinities and vice versa. In gills 7–9, Na–K-ATPase activity was increased more than threefold following a reduction in salinity from 50 to 10 S. Na–K-ATPase activity, expressed as percentage of total ATPase activity, amounted to 60–80% in gills, about 60% in hypodermis and 20–40% in the other organs. Ouabain, a specific inhibitor of Na–K-ATPase activity, reduced serum osmolalities in crabs kept at 9–10 S only when injected into the hemolymph (1 and 5 · 10^-5 M), but had no effect when dissolved in ambient water (10^-4 M). The results obtained underline that crustacean gills are the main organs for ionic regulation, and confirm the hypothesis of the central role of the Na–K-ATPase in active Na uptake as the basic mechanism of hyperregulation in dilute media. Reduction of serum osmolalities following injection of ouabain into the hemolymph confirms previous reports on localization of the sodium pump in the basolateral parts of epithelial cells.     

Interindividual variability in metal status in the shore crab<Emphasis Type="Italic"> Carcinus maenas</Emphasis>: the role of physiological condition and genetic variation

The relations between genetic variation, physiological condition, size and metal status in the shore crab Carcinus maenas (L.) were investigated. Shore crabs were collected at three sites around the island of Funen, Denmark, and carapace width, colour, moulting stage and major ions and protein in the haemolymph were determined. Concentrations of water, cadmium, copper and zinc in midgut glands, muscles and gills were measured, and allozyme variation at 15 loci was studied. Generally, tissue water content and copper, zinc and cadmium concentrations decreased from the moulting stages C₃ over C₄ to D crabs, whereas the opposite was true for copper and zinc concentrations in gills and muscle. However, the water content of the tissues increased with the size of the crabs. Since tissue water contents changed consistently over moulting stages, conclusions on changes in metal concentrations sometimes diverged when expressed on a dry versus wet weight basis. Regarding allozymes, esterase, malate dehydrogenase and phosphoglucomutase were polymorphic. The crabs showed a low level of genetic variability for both polymorphism (0.14) and for heterozygosity (0.07) at the three sites. Allelic frequencies showed very low levels of differentiation among the samples, no evidence of inbreeding and no population subdivision. Multilocus heterozygosity was positively correlated with size. Concentrations of copper and zinc in midgut gland, muscle tissue and gill were not correlated with genotype, whereas a weak correlation between phosphoglucomutase genotypes and concentrations of cadmium in the midgut gland was found.Communicated by M. Kühl, Helsingør     

Entrainment of the larval release rhythm of the crab Rhithropanopeus harrisii (Brachyura: Xanthidae) by cycles in salinity change

Influences of salinity, Na, K, Ca and Mg on Na–K-ATPase activity in the posterior gills of Carcinus maenas (L.) have been investigated with respect to the role of the enzyme in hyperosmotic regulation. K and Mg ions were obligatory for enzyme activity. The dependence on the K concentration can be seen in a saturation curve of the Michaelis-Menten type. Low concentrations of Ca (0.2–3 mM) in the incubation medium strongly inhibited Na–K-ATPase activity. Activities inhibited by Ca could be reactivated to non-inhibited values by the addition of higher amounts of Mg (25 mM). Activity increased along with the salinity of the sea water used as incubation medium up to about 10 S. Here, maximum activity was observed. Further salinity increases of the incubating sea water were inhibitory. Salinity dependence is assumed to be based on Na dependence of the Na-pump. Comparative investigations of the Na–K-ATPase activity and its affinity to sodium in five species of decapod crustaceans indicated that levels of Na–K-ATPase differed in the posterior gills of stenohaline and euryhaline species. The results obtained confirm previous assumptions of a central role of the branchial Na–K-ATPase in hyperosmotic regulation. Properties of the Na–K-ATPase, such as affinity for substrates or dependence on ionic sea water constituents, are kept constant with respect to salinity changes. Modifications due to salinity only concern enzyme amounts especially in the posterior gills. The finding that the Na-pump is localized in basolateral parts of ion-transporting epithelial cells confirms the aforementioned results.     

Phenotype-environment matching in the shore crab (Carcinus maenas)

The shore crab (Carcinus maenas) exhibits a range of carapace pattern polymorphisms, but little is known regarding their function or maintenance. If patterns represent some form of crypsis, then associations between carapace colouration and substrate are expected; to determine whether such relationships exist, frequency of crab morphs and quantity of substrate type were measured from fifteen 10×40 m² quadrats at each of three sites along the southern shore of the Firth of Forth, Scotland. Five thousand one hundred and thirty-seven crabs and 3.6 km of line intercept transect data were collected during a 9-week period. Crab abundance, relative frequency of morphs and substrate type varied significantly among the three sites. Plain crabs were strongly associated with macro-algal substrates whereas patterned crabs were associated with mussel beds. This pronounced phenotype-environment matching, as well as various characteristics of the carapace patterns themselves, suggests that patterned crabs are cryptic on polychromatic backgrounds. The frequency of patterned crabs and the percentage of white pigment on the carapace both declined significantly with carapace width. The loss of pattern coincides with an ontogenetic shift in habitat use and we present evidence to suggest that individual crabs lose their pigment, rather than larger patterned crabs being preferentially removed from the population by predators. Throughout their life history, shore crabs encounter high variation in predation, food supply, and physical habitat; to survive they have evolved a strategy that includes elements of pattern polymorphism, crypsis, ontogenetic shifts, and plastic responses.     

Interactions of 233Pa with tissues of Mytilus galloprovincialis and Carcinus mediterraneus

Uptake of ²³³Pa from sea water by mussels Mytilus galloprovincialis L. and shore crabs Carcinus mediterraneus Csrn. was studied. Results indicate a high contamination factor in digestive tract, gills and skeleton of both investigated species and an increment of the contamination factor in mussel byssus. In spite of this, reproductive system, muscle and hemolymph were only negligibly contaminated. The presence of complexone EDTA in sea water has no effect on uptake of ²³³Pa in the animal tissues tested.     

Variation in the carapace pattern of juvenile Carcinus maenas

Juvenile shore crabs (Carcinus meanas L.) are highly variable with respect to carapace pattern; young crabs often have conspicuous patches of white or other pigment. Adult crabs, in contrast, are drab and usually very similar in pattern. It is probable that the patterns serve to make young crabs cryptic on certain backgrounds. Juvenile crabs of carapace widths lying between 5 and 15 mm were collected from a range of British shores (totalling 48). Pattern incidence was found to relate to two factors: the percentage of weed cover (mainly of fucoids) on the shore, and the amount of mud present. Shores of high weed cover, or with much mud present, had low frequencies of conspicuously patterned crabs. The presence of patterned crabs among the juvenile population, and their absence among adults, could result from differential visual predation removing patterned crabs progressively so that only unpatterned crabs would survive to adulthood. Alternatively, individual crabs could lose their conspicuous patterns with age. Evidence is presented in support of the latter hypothesis.     

The effect of salinity on survival,bioenergetics and predation risk in the mud crabs <Emphasis Type="Italic">Panopeus simpsoni</Emphasis> and <Emphasis Type="Italic">Eurypanopeus depressus</Emphasis>

The effect of salinity on survival, bioenergetics and predation risk was studied in two common mud crabs in the Gulf of Mexico, Eurypanopeus depressus and Panopeus simpsoni. Eurypanopeus survived better at low salinities (the 28-day LC₅₀ of E. depressus was 0.19 PSU compared with 6.97 PSU for P. simpsoni). While low salinity increased energy expenditure and reduced food consumption and absorption, resulting in lower scope for growth, identical responses to salinity occurred in both species. Both species also had similar salinity-dependent patterns of hyper-osmoregulation. Because these physiological mechanisms could not explain differences between the two species in salinity tolerance, we explored the effect of salinity on competition for refugia. Eurypanopeus had higher resource holding potential for refugia, especially at low salinity. As a consequence it had lower predation risk to blue crabs in laboratory experiments. The higher tolerance by E. depressus for low salinities, and greater resource holding potential for refugia may explain why it has a more euryhaline distribution than P. simpsoni.     

Physiological role of branchial carbonic anhydrase in the shore crabCarcinus maenas

Excretion of total CO₂ and uptake of sodium and chloride ions across the branchial epithelium of the posterior gills of the shore crabCarcinus maenas, collected from Kiel Bay (Baltic Sea) in 1989, were measured using isolated perfused gill preparations. Total CO₂ effluxes depended on the HCO ₃ ^- concentration of the internal perfusate in a saturable mode and were inhibited by internally and externally applied acetazolamide at 10^–4 M. Potential differences between hemolymph space and medium did not change significantly during experimental treatments. Neither a bicarbonate gradient (6 mM) directed from the internal perfusate to external bath solution nor symmetrically applied 10^–4 M acetazolamide significantly influenced the influxes of Na⁺ and Cl^–. Results confirmed the role of carbonic anhydrase in CO₂ excretion but called into question the assumed functioning of the enzyme in branchial ion transport processes.     

Competitive role of calcium in sodium transport in the crab carcinus mediterraneus Acclimated to low salinities

Influence of calcium on sodium fluxes was investigated in the brackish-water crab Carcinus mediterraneus Csrn., after activation of sodium regulatory mechanisms, during longterm acclimation in diluted (15.9 S) sea water. The ²²Na outflux constants measured in whole crabs are noticeably lower (0.188 to 0.374h^-1) in diluted sea water enriched by calcium (5.8 to 10.4 mM Ca²⁺/l), than in ordinary diluted sea water (0.545 h^-1). The sodium-outflux constants in hemolymph, gills and muscle show the same trend of slower exchange of ²²Na in calcium-enriched sea water. In ordinary sea water, the total sodium-outflux rate from the hemolymph amounts to 46.31 M Na/g/h, while in calcium-enriched sea water (8.23 mM Ca²⁺/l) it is inhibited, amounting to 13.86 M Na/g/h. Sodium and potassium concentrations of intracellular muscles in diluted sea water enriched with calcium and control diluted sea water are similar. The outflux of intracellular sodium from the muscle amounts to 2.84 M Na/g/h in crabs acclimated to diluted sea water.     

Control of populations of the coral-feeding nudibranch Phestilla sibogae by fish and crustacean predators

Juvenile lesser blue crabs, Callinectes similis Williams, were exposed to a range of salinities for measurement of survival and bioenergetics. Effects of salinity on survival were determined by exposing juvenile crabs to salinity treatments ranging from 0 to 74‰. All crabs survived 21 d of exposure to 5 and 45‰S. The 21 d LC₅₀ values for salinity tolerance (calculated from survival data) were 2.6 and 60.8‰S at low and high salinities, respectively. Energy-budget components and scope for growth were determined for crabs exposed to 2.5, 10, 25, 35 and 50‰S. Energetic absorption rates were highest at 2.5 and 35‰S. Energetic expenditure rates (energy lost to respiration and excretion) were greatest at 2.5‰S, and decreased as salinity increased. Respiration constituted the majority of energetic expenditure at all salinities (92.3% average). Scope for growth was significantly affected by salinity and was highest in crabs exposed to 35‰S. Increased respiration at low salinity may indicate that C. similis incurs greater costs due to osmoregulation. The results of this study indicate that C. similis is capable of surviving and growing in waters with salinities as low as 10‰. Received: 10 January 1997 / Accepted: 11 February 1997     

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     

Cadmium accumulation in the female shore crab Carcinus maenas during the moult cycle and ovarian maturation

 The influence of moulting and ovarian maturation on cadmium accumulation in the tissues of female shore crabs Carcinus maenas exposed to 1 mg Cd l⁻¹ in the water was investigated. Cadmium accumulation in all tissues examined was markedly increased in crabs in the postmoult stages (A and B) compared to crabs in all other moult stages. During the moult cycle, average cadmium accumulation in the midgut gland ranged from 29 μg Cd g⁻¹ dw at premoult stage (D₂) to 589 μg Cd g⁻¹ dw at postmoult stage (A). Average cadmium concentrations in the haemolymph ranged from 0.56 μg Cd ml⁻¹ at intermoult stage (C₄) to 4.6 μg Cd ml⁻¹ at postmoult stage (A), while the gills accumulated from 103 μg Cd g⁻¹ dw in intermoult stage (C₃) to 352 μg Cd g⁻¹ dw in postmoult stage (A). Cadmium concentration in gills and haemolymph was also significantly higher in crabs in late premoult stage (D₃) compared to C₄-crabs, while midgut gland cadmium concentration remained elevated in C₁- and C₃- intermoult stages relative to C₄. During ovarian maturation the cadmium accumulation in midgut gland, gills, ovaries and haemolymph decreased. Average cadmium concentration in the midgut gland decreased from 63 μg g⁻¹ dw in ovarian Stage I to 19 μg g⁻¹ dw in ovarian Stage VI. The same pattern was observed for gills, haemolymph and ovaries. The present study demonstrates that cadmium accumulation in the female shore crab strongly depends on the physiological status of the animal. A possible association between physiological calcium requirements and cadmium accumulation during moulting is discussed. Received: 20 January 2000 / Accepted: 20 July 2000     

Voltage-clamp measurements on single split lamellae of posterior gills of the shore crab Carcinus maenas

Single split lamella preparations of posterior gills of low-salt adapted shore crabs Carcinus maenas (collected from the Kiel Bay, Baltic Sea in 1991) were mounted in a micro Ussing-chamber. With NaCl salines on both sides we found an outside positive potential difference (PD) of 6.6±1.3 mV, a short-circuit current (I_sc) of-240±65 Acm^-2 and a resistance (R_t) of 25±3 cm² (n=8). Substitution of Cl^- (gluconate) on both sides of the preparation resulted in a decrease of I_sc by more than 90% at constant R_t. I_sc disappeared and R_t increased after substitution of Na⁺ (choline). When ouabain (2 mmol l^-1) was applied to the internal NaCl-saline, I_sc decreased and R_t remained unchanged. Internal addition of 0.1 mmol l^-1 acetazolamide left I_sc and R_t unaffected. Application of amiloride to the external NaCl saline resulted in a increase of both inward negative I_sc and R_t. The dose dependence of the diuretic showed a maximal effect between 50 and 200 mol l^-1 with a half-maximal blocker concentration (K_AMI) of ca. 10 mol l^-1. The results show that the split lamella preparation of posterior gills of C. maenas is a low resistance epithelium which is able to effect a massive, electrogenic and coupled absorption of Na⁺ and Cl^-.     

Calanoid copepod eggs in sea-bottom muds. IV. Effects of some environmental factors on the hatching of resting eggs

During cadmium uptake by the shore crab Carcinus maenas (L.), cadmium appears in both haemolymph and urine within 1 h, and comes into equilibrium with the haemolymp after some 60 h. The equilibrium concentration for haemolymph cadmium is always considerably less than that of the external medium, whether the latter is close to background level or at the artificially high concentration of 2.2 mg l^-1. Despite earlier evidence that cadmium entering the haemolymph is mainly bound to protein, current results show most of this to be freely labile and quickly displaced by cadmium taken up subsequently. There is some evidence to suggest that cadmium is passed on from the haemolymph to the hepatopancreas. A large proportion of whole-body cadmium is adsorbed onto the exoskeleton and gills. The comparatively rapid efflux of ¹⁰⁹Cd from loaded crabs over a period of 1 week is mainly due to desorption of this cadmium from the outer body surface.