Kinetics of uranium uptake by the crab Pachygrapsus laevimanus and the zebra winkle Austrocochlea constricta

The crab Pachygrapsus laevimanus and the zebra winkle Austrocochlea constricta were exposed for 40 d to uranium (1.5 to 10 mg l^-1) in continuous-flow sea water in separate starved and fed treatments, and the kinetics of uranium bioaccumulation were estimated from an exponential model. Starved and fed crabs took up U at a similar rate, which suggests that sea water was the major source of U to the crab; the fed crabs excreted U more rapidly than the starved crabs and this led to a lower net uptake of U by fed crabs. Fed and starved winkles took up U at similar rates and excreted it at similar rates, so the sea water was also the major source of U to winkles. Crabs took up more U than winkles; the concentration factors were 7 to 18 and 4, respectively. Uranium turnover was quite slow for both species (11 to 36 d) as it was also for winkle shells (6 d); this suggests that the rate-limiting processes which control turnover are biological (e.g. growth or tissue replacement) or physical (e.g. diffusion into the shell) rather than chemical (e.g. precipitation, adsorption or exchange). There was no effect of increasing U concentration in water on the U kinetic parameters.     

Studies on the uptake of cadmium by the crab Carcinus maenas in the laboratory. I. Accumulation from seawater and a food source

The crab Carcinus maenas (L.) was exposed to radioactively labelled cadmium dissolved in seawater at concentrations of 0.1, 1 and 10 ppm, the latter concentration being toxic to the crabs (50% mortality after 12.3 days). Net accumulation of cadmium from solution was proportional to the level and time period of cadmium exposure. Total absorbed cadmium levels reached 0.0043 and 0.0412 mg Cd g^-1 dry weight after 40 days exposure to 0.1 and 1 ppm Cd, respectively, and 0.1115 mg Cd g^-1 dry weight after 12.3 days average exposure to 10 ppm Cd. The highest tissue concentration was found in the midgut gland, reaching 0.786 mg Cd g^-1 dry weight after 12.3 days average exposure to 10 ppm Cd. The midgut gland only contained about 10% of the total cadmium absorbed from solution, while the exoskeleton contained the bulk of obsorbed cadmium (59 to 80%) probably passively adsorbed onto the surface. When cadmium was absorbed by the crabs from a food source, the midgut gland contained 16.9% of the total absorbed cadmium whereas the exoskeleton now contained only 22.2%. Ten percent of the cadmium available in the food source (Artemia salina) was accumulated by the crabs. When placed in cadmium-free seawater, crabs that had accumulated cadmium from solution lost 69% of the absorbed cadmium in 10 days, mostly from the exoskeleton which lost 78% of its original absorbed cadmium concentration.     

Myosin ATPase activity in an estuarine decapod crustacean,Scylla serrata,as a function of salinity adaptation

The myosin ATPase activity of the flexor muscle of an estuarine crab, Scylla serrata, was studied in relation to salinity adaptation. The enzyme is activated more by calcium than by magnesium; it exhibits maximum activity at pH 9.0, and substrate inhibition above 0.5 mM ATP. The enzyme activity increases in crabs adapted to higher salinities. The enzyme from normal (70% sea water) crabs shows two pH optima; one at pH 7.0, the other at pH 9.0. The neutral optimum shifts to pH 6.0 upon adaptation to full strength sea water, but disappears upon adaptation to 25% sea water. The enzyme from normal crabs shows an optimum at 30 °C; adaptation to full strength sea water raises this optimum to 38 °C, whereas adaptation to 25% sea water decreases it to 24 °C. These changes are discussed in relation to estuarine conditions.     

Influence of physiological condition on cadmium transport from haemolymph to hepatopancreas inCarcinus maenas

Accumulation and binding of cadmium in the tissues of the shore crabCarcinus maenas (L.) were investigated in a series of laboratory experiments. The relation between the physiological condition of individual crabs and their ability to transport cadmium from haemolymph to hepatopancreas was specifically addressed. Cadmium was removed from the haemolymph with a half-life of approximately 10 h, and half of the cadmium removed from the haemolymph was taken up in the hepatopancreas. The efficiency with which individual crabs transported cadmium from the haemolymph to the hepatopancreas was strongly related to physiological parameters, such as concentrations of calcium and magnesium in the hepatopancreas and haemolymph volume and haemolymph protein concentration. The transport of cadmium from the haemolymph to the hepatopancreas was saturated in crabs exposed to more than 2 to 4 mg Cd l^–1 in the sea water. In the hepatopancreas of unexposed crabs, cadmium was bound mainly in the insoluble tissue fraction (40%) and in the protein fraction with a molecular weight of approximately 6000 D (50%). Exposure to 0.25 to 1.5 mg Cd l^–1 for 2 w led to dose-dependent increases of the amounts of cadmium bound in the high-molecular weight protein fraction and in the insoluble tissue fraction. Modes of internal cadmium transport and accumulation in tissues and variability in physiological parameters are discussed.     

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     

Combined effects of copper and cadmium on Chlorella pyrenoidosa H.Chick: subcellular accumulation,distribution, and growth inhibition

Disposal of waste into aquatic ecosystems may cause microalgae to be exposed to various metals, e.g. copper and cadmium. The effects caused by combinations of metals may be more serious. Evaluations of subcellular fate, bioaccumulation, and biological effects of metals on aquatic organisms are generally derived from experiments with individual metals. The present study aims to evaluate the effects of exposure of Chlorella pyrenoidosa to copper and cadmium in combination on subcellular accumulation, distribution, and growth. The algae were exposed for 72 h to copper at concentrations of 13 – 25 µmol L^?1, cadmium at about 6 µmol L^?1, and combinations thereof. The levels of copper and cadmium in subcellular organelles, heat-denaturated protein, metal-rich granules, and heat-stable protein were determined by atomic absorption spectrometry. Exposure of C. pyrenoidosa to copper and cadmium in combination inhibited growth more strongly than copper and cadmium individually. Highest accumulation was observed in metal-rich granules and heat-stable proteins. Administration of both metals in combination affected their subcellular distribution: copper was mainly distributed into the metal-rich granules (70%–80%) and heat-stable proteins (9%–24%), cadmium in the metal-rich granules (88%–98%).     

The common mussel Mytilus edulis as an indicator of pollution by zinc,cadmium, lead and copper. I. Effects of environmental variables on uptake of metals

The net uptake of zinc, cadmium, lead and copper by the common mussel Mytilus edulis (L.) exposed to different conditions was investigated with a view to using this species as an indicator of contamination of the marine environment by these metals. The variables studied were season, position of the mussel in the water column, water salinity, water temperature, and the simultaneous presence of all four metals. Each of these 5 variables affected the net uptake of some or all of the metals studied under some conditions. Seasonal variation in concentrations of zinc, cadmium and copper was found in samples collected at three separate locations. The relationship of seasonal variation to tissue weight and absorption route of the metals is discussed. Near to freshwater inputs of trace metals, the concentrations of zinc, cadmium and lead in mussels were found to vary according to the depth at which the mussels were collected; in summer when freshwater run-off is less, this effect was absent. Low salinities did not affect the net uptake of zinc by mussels, but increased the net uptake of cadmium and decreased that of lead. Low temperatures had no effect on the net uptake of zinc or lead; the net uptake of cadmium was unaffected by low temperatures at high salinities but was decreased by low temperatures at low salinities. The presence of the other metals had no effect on the individual net uptake of either zinc, cadmium or lead. A sampling programme was devised to eliminate the effects of these environmental variables and to allow the use of M. edulis as an indicator of zinc, cadmium and lead in marine and estuarine environments. In contrast to the other metals, the net uptake of copper by the mussel was extremely erratic, and was affected by salinity and temperature changes and by the presence of the other metals and changes in their relative concentrations. The effects of other metals on the net uptake of copper cannot be easily eliminated or allowed for; it is, therefore, suggested that the mussel should not be used as an indicator of copper in the marine environment.     

Comparison of osmoregulation between Adriatic and North Sea Carcinus

In two spatially isolated species — North Sea Carcinus maenas L. and Adriatic Carcinus mediterraneus Czerniavsky —osmoconcentration in hemolymph and Na-transport were investigated. The experiments were performed on crabs which had been long-term acclimated to sea water (1190 mOsmol/l) and to 50% sea water, and on individuals immediately after transfer from sea water to 50% sea water, until new steadystate conditions had been reached. About 10 to 12 h after transfer, a new steady state of total osmoconcentration as well as of the concentrations of the most representative cations Na, K, Ca, and Mg was attained. The hemolymph contents of Na and total osmotically active substances of the North and Adriatic Sea populations are significantly different when they are long-term acclimated to sea water and to 50% sea water. Total sodium fluxes, as calculated from influx and outflux experiments, are in the range 12.1 to 15.2 M Na/g/h in both species when acclimated to sea water. In 50% sea water, sodium fluxes are retarded, especially in C. maenas (6.6 M Na/g/h). This value is significantly lower than the corresponding value in C. mediterraneus (11.4 M Na/g/h). Differences are discussed on the basis of adaptation of the populations to their different environments.     

Toxicity of zinc,cadmium and copper to the shrimp Callianassa australiensis. III. Accumulation of metals

Callianassa australiensis (Dana) that survived 14 d acute lethality studies were analysed to determine the concentrations of zinc, cadmium and copper in the whole shrimp and in various parts of the body. Using regression analysis, the influence of each metal upon the uptake of the others was studied. Zinc and cadmium appeared to enhance the uptake of each other. In a mixture of zinc and copper, the uptake of zinc was enhanced and that of copper was inhibited. In a mixture of cadmium and copper, the uptake of copper was inhibited by the presence of cadmium, but cadmium uptake was unaffected in the presence of copper. In a mixture of all three metals, similar effects were observed except that zinc and copper, occurring together, appeared to have no effect upon cadmium uptake. Additional 14 d experiments with cadmium suggested that accumulation of this metal was a function of metal concentration in the water and of duration of exposure. The whole shrimp cadmium concentration also appeared to be a function of the size of the shrimp. The variation in concentration factors is described and the need for further research on the effects of combinations of metals on various organisms is emphasized.     

Dispersal,population structure and burrow shape of Ocypode cursor

The distribution of the sand crab Ocypode cursor (L.), as indicated by the number of burrows, was studied for 2 years in a 50×50 m sand beach area in northern Israel. A definite relationship was established between the distribution pattern from the seashore inwards towards the sand dunes, and the degree of sand moisture as it changed seasonally. During autumn, more crabs were found at a distance of 15 to 25 m from the sea where sand moisture was about 14%. At the beginning of winter crabs dispersed evely, disappearing with advancing winter. Crabs reappeared in spring, although in smaller numbers, dispersing in a pattern similar to that in autumn. At the beginning of summer and later on, more crabs appeared and concentrated closer to the sea (5 to 10 m). The population structure was analysed directly by measuring the crab's dimensions, and indirectly by counting burrows and measuring the diameter of their openings. Direct analysis revealed two distinct sizeage groups: smaller crabs 0.5 to 3 cm long, and larger ones over 4 cm long. The smaller burrows were inhabited by the first group and were mostly found closer to the sea; the second group was found more landwards. Three main types of burrow shapes are described.     

Effects of size upon salinity tolerance and volume regulation in the hermit crab Pagurus bernhardus

Whereas small Pagurus bernhardus (L.) occur in large numbers in the intertidal areas of rocky shores, large hermit crabs living in whelk shells are most uncommon, although they are easily collected by sublittoral dredging. Experiments were carried out to assess the salinity tolerance, and the ability to regulate body volume in a dilute medium, of both small and large hermit crabs. Large, offshore hermit crabs are significantly less tolerant of 60% sea water (100%=34 S) than small, littoral specimens. Small littoral crabs increase in weight by about 15% in the first hour in 60% sea water, but increased urine output prevents swelling thereafter; indeed, there is considerable loss in weight, presumably caused by solute loss. Large, offshore, crabs increase in weight by about 15% in the first hour, thus exhibiting a higher permeability to water than small crabs, and continue to increase in weight, since urine output is not sufficient to cope with the water load. Dissections suggest that insufficient release of urine causes the low urine output in large hermit crabs, rather than inadequate production of primary urine. Calculations indicate that the nephropores of large hermit crabs are too small to allow a sufficiently high urine output to remove a large water load, unless greater pressures are developed inside the excretory systems of large hermit crabs than occur in those of small littoral individuals. In view of the soft, flexible nature of the walls of the excretory system and abdomen of P. bernhardus, it seems unlikely that high pressures could be maintained within the excretory system. It is concluded that the stenohalinity of large, offshore hermit crabs may exclude them from the littoral zone, or may be the result of exclusion by other factors, since there will be no selective advantage for large hermit crabs in maintaining the degree of euryhalinity possessed by the small littoral crabs, if the large crabs are confined to the sublittoral zone.     

Some beavioral characteristics related to food supply and soil texture of burrowing habitats observed on Uca vocans vocans and U. lactea perplexa

From the observations on two species of fiddler crabs, droving is conspicuous in Uca vocans vocans but does not occur in U. lactea perplexa. U. vocans come down to feed at the water's edge with a richer food supply and fine substrate particles. Droving is advantageous to increase feeding efficiency in this species which holds burrows on a relatively sandy habitat where the food supply is less rich. In U. lactea, which burrows at higher levels, on the other hand, the food supply and soil texture of their burrow area are not significantly different from those at the water's edge. Home range relationships of both species are consistent with the occurrence or not of droving behavior and depend on food productivity of burrowing habitats.     

Field and experimental studies on cadmium in the edible crab Cancer pagurus

The distribution of cadmium within captive crabs (Cancer pagurus) exposed experimentally to cadmium-contaminated food and water is described and illustrated by triangular diagrams. Crabs from the Orkney Islands (Scotland) are known to contain relatively high levels of cadmium (up to 62 g g^-1 wet wt) in the hepatopancreas. The distribution of cadmium between the hepatopancreas, gonad, gill, carapace, claw muscle, heart, and haemolymph, is described in crabs collected during 1978, and compared with similar data from crabs exposed to cadmium for ca. 300 d (September 1978 – June 1979) experimentally. It is concluded that the dominant uptake route of cadmium to Orkney crabs is through their diet.     

The Influence of Temperature and Salinity Upon the Acute Toxicity of Heavy Metals to the Banana Prawn (Penaeus merguiensis de Man)

Bioassays were conducted to determine the effects of temperature and salinity on the acute toxicity of mercury, copper, cadmium, zinc, nickel and lead to juvenile banana prawns (Penaeus merguiensis de Man). Tests were conducted at all combinations of 35, 30 and 20°C with 36 and 20° salinity over 96 h. The general rank order of metal toxicity was Hg > (Cu, Cd, Zn) > Ni > Pb. The toxicity of all metals increased with increased temperature. This was most noticeable in the high salinity treatments, particularly for copper and zinc. Salinity appeared to influence the toxicity of all metals tested although significant differences were only found for copper and lead at 20°C. At this temperature prawns were markedly more susceptible to both metals in low salinity sea water. The data are compared with lethal concentrations found for other crustaceans and are discussed in relation to existing water quality criteria.     

Effects of time of year and limb removal on rates of ecdysis of eyed and eyestalkless fiddler crabs,Uca pugilator

The incidence of ecdysis among monthly samples of female Uca pugilator, both intact and eyestalkless, was determined from May, 1974 through April, 1975. Among both the intact and eyestalkless crabs the percentages of the crabs that underwent ecdysis varied during the year. During the height of the breeding season in particular, ecdyses were rare among the eyestalkless crabs and absent among the intact ones. The effects of limb removal, either followed by or preceded by eyestalk removal, on the rate of ecdysis were also determined. Eyestalkless crabs from which 4 walking legs were removed either the day before or the day after their eyestalks had been removed underwent ecdysis at a slower rate than crabs lacking only eyestalks, but faster than crabs lacking only 4 walking legs. The high incidence of precocious ecdyses induced by removing large numbers of limbs from eyed crabs appears to be due to the number of limbs removed and not to the weight or volume of limb tissue removed.     

Reproductive isolation with little genetic divergence between Urticina (=Tealia) felina and U. eques (Anthozoa: Actiniaria)

There has long been doubt as to whether there are one or two British species of the sea anemone genus Urticina. In the present study, populations of both putative species, which occur sympatrically in the Isle of Man, have been compared by electrophoresis of isozymes and nematocyst analysis. The analysis of fourteen isozyme loci exhibited a high genetic identity between the two taxa (I=0.907), but four loci showed significant differences in gene frequencies; thus indicating reproductive isolation. The populations displayed highly significant differences in the sizes of the various nematocyst types. The individual analysis of data for each type of nematocyst revealed that the basitrics from the actinopharynx and from the tentacles contribute most to the observed difference. Consequently, U. felina (L.) and U. eques (Gosse) are assumed to be valid species, as suggested by Manuel (1981). The mean heterozygosities for each species (0.410 and 0.436, respectively) are the highest reported for cnidarians.     

Effects of copper and cadmium on osmoregulation and oxygen consumption in two species of estuarine crabs

Green crabs (Carcinus maenas) and rock crabs (Cancer irroratus) were exposed to various concentrations of copper as cupric chloride (CuCl₂ · 2 H₂O), and cadmium as cadmium chloride (CdCl₂ · 21/2 H₂O) for 48 h. The exposures were conducted at 5 different salinities. At the end of each exposure period, tests of blood-serum osmolality and gill-tissue oxygen consumption were performed. Copper-exposed crabs exhibited loss of osmoregulatory function with increasing copper concentration until normally hyperosmotic serum became isosmotic with the surrounding medium. Cadmium elevated greencrab serum above its normal hyperosmotic state. Copper had no effect on gill-tissue oxygen consumption; however, cadmium reduced the rate of oxygen consumption in both species tested.     

The common mussel Mytilus edulis as an indicator of trace metals in Scandinavian waters. I. Zinc and cadmium

Concentrations of zinc and cadmium in whole soft parts of mussels, Mytilus edulis (L.), collected from 54 locations in Scandinavian waters were determined. Local variations in concentrations of the two metals found in samples taken close to industrial sources of zinc and cadmium confirmed the ability of the mussel to act as an accurate indicator of pollution by these metals over the entire range of salinities in which this species can exist. In addition, offshore samples remote from industrial discharges revealed higher concentrations of zinc and cadmium in mussels from lowsalinity areas (Gulf of Finland, Southern Bothnian Sea, Baltic proper) than in those from high-salinity areas (Kattegat, Eastern Skagerrak). Major decreases in metal concentrations present in the mussel were apparent in the regions of the Sound and the Great Belt, which are areas of rapid salinity change due to mixing of Baltic water with water from Kattegat. Comparison of these results with those reported for zinc and cadmium in water throughout the study area suggested the existence of a higher biological availability of these metals in regions of low salinity; possible reasons for this are discussed. Data from the present survey are also compared to those reported for mussels taken elsewhere. This comparison reveals the Baltic area to be considerably polluted by trace metals; in contrast, the waters of Kattegat and skagerrak are considered to be relatively unpolluted.     

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.     

The effect of chelating agents on the uptake and accumulation of cadmium by Mytilus edulis

The uptake, storage and excretion of cadmium by the common mussel Mytilus edulis (L.) has been studied at sub-lethal concentrations using the radioactive isotope ^115mCd as a marker. After an initial lag period, the uptake at low concentrations in sea water is linear with time and directly proportional to the sea water concentration, with a maximum concentration factor of 165 at 0.7 g Cd/ml sea water. A decrease occurs at higher concentrations indicating saturation of the available binding capacity. Prior complexation of the cadmium with either EDTA, humic and alginic acids or pectin doubles both the rate of accumulation and the final tissue concentrations (order: kidneyviscera>gillsmantle>muscle) and eliminates the lag period, suggesting that ionic cadmium must first be complexed before uptake can occur. A mechanism for this effect, which may involve thionein, is described. The rate of excretion of cadmium is 18 times slower than that of uptake, with the major route via the kidney but not via the byssal threads as with particulate iron. The need to detoxify and store cadmium by an immobilization mechanism is a consequence of this slower rate of elimination.