Effect of sexual maturation on the tissue biochemical composition of<Emphasis Type="Italic"> Octopus vulgaris</Emphasis> and<Emphasis Type="Italic"> O</Emphasis>.<Emphasis Type="Italic"> defilippi</Emphasis> (Mollusca: Cephalopoda)

Changes in the protein, lipid, glycogen, cholesterol and energy contents, total amino acid and fatty acid profiles of Octopus vulgaris and O. defilippi tissues (gonad, digestive gland and muscle) during sexual maturation (spermatogenesis and oogenesis) were investigated. Both species showed an increase of amino acids and protein content in the gonad throughout sexual maturation (namely in oogenesis), but allocation of these nitrogen compounds from the digestive gland and muscle was not evident. The major essential amino acids in the three tissues were leucine, lysine and arginine. The major non-essential amino acids were glutamic acid, aspartic acid and alanine. With respect to carbon compounds, a significant increasing trend (P<0.05) in the lipid and fatty acid contents in the three tissues was observed, and, consequently, there was also little evidence of accumulated lipid storage reserves being used for egg production. It seems that for egg production both Octopus species use energy directly from food, rather than from stored products. This direct acquisition model contrasts with the previous model for Octopus vulgaris proposed by ODor and Wells (1978: J Exp Biol 77:15–31). Most of saturated fatty acid content of the three tissues was presented as 16:0 and 18:0, monounsaturated fatty acid content as 18:1 and 20:1 and polyunsaturated fatty acid content as arachidonic acid (20:4n-6), eicosapentaenoic acid (20:5n-3) and docosahexaenoic acid (22:6n-3). Though cholesterol is an important precursor of steroid hormones, this sterol content exhibited variations that do not seem to be related with the maturation process. Moreover, significant differences (P<0.05) were obtained between genders, suggesting that perhaps there is a greater physiological demand for cholesterol during spermatogenesis than oogenesis. If the component sterols of octopus are of a dietary origin, considerable variation in the cholesterol content between species might be expected on the basis of the sterol composition of their prey. The glycogen reserves increased significantly in the gonad and decreased significantly (P<0.05) in the digestive gland and muscle of O. vulgaris (these trends were not evident in O. defilippi). Glycogen may play an important role in the maturation process and embryogenesis of these organisms, because carbohydrates are precursors of metabolic intermediates in the production of energy. It was evident that sexual maturation had a significant effect upon the gonad energy content, but the non-significant energy variation (P>0.05) in the digestive gland and muscle revealed no evidence that storage reserves are transferred from tissue to tissue. The biochemical composition of digestive gland and muscle may not be influenced by sexual maturation, but rather by other biotic factors, such as feeding activity, food availability, spawning and brooding.Communicated by S.A. Poulet, Roscoff     

Oxygen consumption and ammonia excretion of<Emphasis Type="Italic"> Octopus vulgaris</Emphasis> (Cephalopoda) in relation to body mass and temperature

The common octopus, Octopus vulgaris Cuvier, is of great scientific and commercial importance and its culture is becoming an area of increasing interest. In this study, the combined effects of temperature (T) and body mass (M) on the routine oxygen consumption rate (R) and ammonia excretion rate (U) in O. vulgaris were quantified. The experiments were conducted in a closed seawater system, and great care was taken to reduce handling stress of the animals. Temperature, salinity, pH and ammonia, nitrite, nitrate and phosphate concentrations were monitored and controlled during the experiment. The following predictive equations were evaluated: at temperatures between 13°C and 28°C and at temperatures between 15.5°C and 26°C (T_a is degrees Kelvin and M in gram). O/N ratios showed that O. vulgaris has a protein-dominated metabolism. No significant relationship between the O/N ratio and body mass or temperature was found. Sex had no significant effect on the oxygen consumption rate or on the ammonia excretion rate. For other octopod species, the dependence of metabolic rate on temperature does not differ with that for O. vulgaris.Communicated by O. Kinne, Oldendorf/Luhe     

Petroleum hydrocarbons in the marine bivalve Venus verrucosa: accumulation and cellular responses

Cellular and subcellular responses of the marine burrowing bivalve Venus verrucosa collected from the north-eastern coastline of Malta from January to June 1985, after exposure to petroleum hydrocarbons (PHC) were investigated. After long-term exposure to 100 gl^-1 of water-accomodated fractions (WAF) of crude oil, PHC were found to accumulate most rapidly in the digestive gland and then in the gills, with saturation levels being reached within 100 d of exposure in both cases. PHC accumulation, both in the mantle and muscle tissues, was more gradual and consistent throughout the whole exposure period. After 150 d of exposure, the digestive cells of the digestive gland were significantly reduced in height (atrophy) and exhibited reduced lysosomal membrane stability. After 144 h of exposure to higher concentration of PHC (820 and 420 gl^-1), several cytological effects were recorded, including an increase in cell volume and activity of gill mucocytes as well as in the number of haemocytes in gill blood sinuses. There was also evidence of damage to the epithelial lining of the foot, stomach and style sac and marked atrophy of the digestive cells of the digestive gland. The significance of such responses is discussed.     

Effect of temperature on specific dynamic action in the common octopus,<Emphasis Type="Italic"> Octopus vulgaris</Emphasis> (Cephalopoda)

Feeding causes an increase of metabolic rate, which initially escalates rapidly, reaches a peak value and then gradually declines to the pre-feeding rate. This phenomenon, termed specific dynamic action (SDA), reflects the energy requirements of the behavioral, physiological and biochemical processes that constitute feeding. The effect of temperature on SDA of the common octopus, Octopus vulgaris, was evaluated, by measuring the temporal pattern of the oxygen consumption rates of octopuses, after feeding, at two constant temperatures, 20°C and 28°C. At 20°C, the relative increase in the oxygen consumption rate after feeding (relative SDA) was significantly greater than at 28°C. The peak of the relative SDA occurred 1 h after feeding, and it was 64% at 20°C and 42% at 28°C. However, the SDA absolute peak, SDA duration (9.5 h) and SDA magnitude (the integrated postprandial increase in oxygen uptake) did not differ significantly between the two temperatures, indicating that the energetic cost of feeding was the same at both temperatures. The SDA response in O. vulgaris was much faster than it was in polar species, which have extended SDA responses due to low temperatures, and was also relatively fast in relation to the response in other temperate species, which is probably connected to the remarkably high growth rates of the species. A possible explanation of the observed summer migration of large octopuses from shallow to deeper areas is given, based on the effect of temperature on the energetic requirements of octopuses.     

Concentration and distribution of heavy metals in tissues of two cephalopods,Eledone cirrhosa and Sepia officinalis,from the French coast of the English Channel

The concentrations of 11 heavy metals (Ag, Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb, V and Zn) were measured in the tissues (digestive gland, branchial hearts, gills, digestive tract, kidney, genital tract, muscle, skin, shell) of the two cephalopods Eledone cirrhosa (d'Orb.) and Sepia officinalis (L.) collected from the French coast of the English Channel in October 1987. The tissues of both species displayed a similar pattern of heavy-metal accumulation: the digestive gland, branchial hearts and kidney were the major sites of concentration for all 11 metals; the digestive gland accumulated silver, cadmium, cobalt, copper, iron, lead and zinc, the branchial hearts high concentrations of copper, nickel and vanadium, and the kidney high concentrations of manganese, nickel and lead. The digestive gland, which constituted 6 to 10% of the whole-animal tissue, contained >80% of the total body burden of Ag, Cd and Co and from 40 to 80% of the total body burden of the other metals. The ratios between heavy metal concentrations in the digestive gland and those in the muscle separated the elements into three groups, those with a ratio 10 (Cr, Mn, Ni, Pb, V, Zn), those with a ratio >10 to <50 (Co, Cu, Fe), and those with a ratio 50 (Ag, Cd). The digestive gland of cephalopods (carnivorous molluscs whose age can be easily calculated with great accuracy) would seem to constitute a good potential indicator of heavy metal concentrations in the marine environment.     

Shelter selection of the spiny lobster <Emphasis Type="Italic">Palinurus elephas</Emphasis> under different levels of <Emphasis Type="Italic">Octopus vulgaris</Emphasis> predation threat

The skill of recognizing and reacting to predators is often based on a learned component. Few studies have examined the role of learning in spiny lobster anti-predator behavior. We investigated whether European spiny lobster (Palinurus elephas) shelter selection is influenced by olfactory stimuli released by one of the most common lobster predators, the common octopus (Octopus vulgaris), and whether the behavioral response to octopus chemical stimuli is innate or influenced by experience. In experimental arenas, we conditioned wild-caught lobsters with three levels of predation threat: no threat, with no predator–prey interaction; medium threat, with odor and visual predator cues only; high threat, active predation risk. We subsequently tested the shelter choice of the conditioned lobster under different experimental conditions: (1) shelter plus seawater; (2) shelter plus seawater plus chemical octopus cue. Our results showed significant differences in mean shelter occupancy with conditioning level. We conclude that P. elephas individuals use chemosensory systems in predator-avoidance mechanisms. Moreover, lobsters subject to a training period of high-level predation threat were able to learn the octopus chemical stimuli and treat its odor as a cue related to predation risk. The findings relative to the spiny lobster learning abilities could be an important tool for future management of lobster populations, e.g., by re-introduction of reared juveniles, which have not yet experienced predation.     

Ontogenetic changes in digestive enzyme activity of the spiny lobster,<Emphasis Type="Italic"> Jasus edwardsii</Emphasis> (Decapoda; Palinuridae)

Digestive enzyme profiles of puerulus, post-puerulus, juvenile and adult stages of the spiny lobster Jasus edwardsii Hutton were determined in order to identify ontogenetic changes in digestive capabilities and assess capacity to use dietary components and to exploit diets to meet nutritional requirements. Juvenile and adult lobsters exhibited a diverse range of enzymes, suggesting they can exploit varied diets. Proteolytic activity of trypsin, chymotrypsin and carboxypeptidase A (1.86–3.70 U mg^–1 digestive gland protein) was significantly higher than carbohydrase activity of amylase, -glucosidase, cellulase and chitinase (0.0014–0.38 U mg^–1 digestive gland protein), thus showing that dietary protein was more important than carbohydrate and that the lobster is carnivorous. These conclusions are consistent with other studies that found spiny lobsters to feed predominantly on crabs, bivalves, ophiuroids and sponges. Lipase activity (0.371 U mg^–1 digestive gland protein) was also relatively high, thus showing the importance of dietary lipid. Total activities (units per digestive gland) of every enzyme assayed increased significantly with lobster carapace length. There were few significant ontogenetic trends in specific enzyme activity (U mg^–1 digestive gland protein). Amylase and laminarinase specific activity was significantly higher in small lobsters than large lobsters (regression analyses, respectively, F_(1,23)=9.84, P=0.005; F_(1,11)=19.65, P=0.001), suggesting that carbohydrates including laminarin are more important in the diet of small juveniles. Trypsin, amylase and lipase activities were detected in all non-feeding puerulus stages, suggesting that feeding is not a cue for digestive enzyme production in J. edwardsii. Significant variations in total and specific activities of amylase (F_(1,3)=14.2, P=0.00; F_(1,3)=14.2, P=0.00) and trypsin (F_(1,3)=8.8, P=0.00; F_(1,3)=21.41, P=0.00) and a declining trend in lipase total activity between non-feeding puerulus stages suggests that they may be hydrolysing endogenous energy reserves to sustain their onshore swimming activity prior to settlement. Profiles suggest carbohydrate and lipid are utilised first, followed by protein. Consistently high levels of lipase in all puerulus stages (0.24–0.7 U digestive gland^–1; P>0.05) confirm the importance of lipid as a major energy substrate.Communicated by G.F. Humphrey, Sydney     

Experimental evaluation of the energy balance in Octopus vulgaris, fed ad libitum on a high-lipid diet

A complete energy balance equation was estimated for the common octopus Octopus vulgaris at a constant temperature of 20°C, fed ad libitum on anchovy fillet (Engraulis encrasicolus). Energy used for growth and respiration or lost with faeces and excreted ammonia was estimated, along with total energy consumption through food, for six specimens of O. vulgaris (with masses between 114 and 662 g). The energy balance equation was estimated for the specimens at 10-day intervals. During each 10-day interval, food consumed, body mass increase and quantity of faeces voided were measured. The calorific values of octopus flesh, anchovy flesh and faeces were measured by bomb calorimetry. Oxygen consumption and ammonia excretion rates were monitored for each specimen during three 24-h experiments and daily oxygen consumption and ammonia excretion were estimated. It was found that 58% of the energy consumed was used for respiration. The amount of energy invested in somatic and gonadal growth represented 26% of the total energy budget. The energy discarded through faeces was 13% of consumed energy. The estimated assimilation efficiency (AE) values of O. vulgaris feeding on anchovy (80.9–90.7%) were lower than the AE values estimated for other cephalopod species with different diets of lower lipid content such as crabs or mussels. Specific growth rates (SGR) ranged 0.43–0.95 and were similar to those reported for other high-lipid diets (bogue, sardine) and lower than SGR values found for low-lipid, high-protein diets (squid, crab, natural diet). Ammonia excretion peak (6 h after feeding) followed the one of oxygen consumption (1 h after feeding). The values of atomic oxygen-to-nitrogen (O:N) ratio indicated a protein-dominated metabolism for O. vulgaris.     

Diet of<Emphasis Type="Italic"> Octopus vulgaris</Emphasis> in False Bay,South Africa

The diet of Octopus vulgaris was analysed using instantaneous daytime observations, midden counts, and stomach contents and a total of 39 prey species were identified. From stomach contents, the most important prey species were Plagusia chabrus (64.6% IRI, index of relative importance) and Haliotis midae (21.6% IRI). Crustaceans were the most frequently found prey group in octopus stomachs (63.6% frequency of occurrence), followed by molluscs (37.6%), teleosts (11.2%), and polychaetes (10.8%). Prey size and diversity increased with increasing octopus size. From middens, the mean shell lengths of H. midae consumed by small, medium, and large O. vulgaris were 53.3, 72.6, and 86.0 mm, respectively. Compared with stomach contents, midden counts were 3 times higher for shelled molluscs, but 5 times lower for crustaceans and soft-bodied organisms. Similarly, instantaneous daytime observations were 3 times higher for shelled molluscs, but 5 times lower for crustaceans and 2 times lower for soft-bodied organisms.Communicated by G.F. Humphrey, Sydney     

Behavioural and physiological responses of the Norway lobster,<Emphasis Type="Italic"> Nephrops norvegicus</Emphasis> (Crustacea: Decapoda), to sulphide exposure

The Norway lobster, Nephrops norvegicus, is moderately tolerant of sulphide [the lethal time to 50% mortality (LT₅₀) was 22.5 h when exposed to 500 M sulphide] but, whenever possible, it attempts to avoid the presence of sulphide in its immediate vicinity. Any sulphide entering the animal is oxidized to thiosulphate, which accumulates in the haemolymph and in the tissues. During exposure to low concentrations of sulphide, the rate of oxygen consumption is maintained or even enhanced even though the lobsters become quiescent. The apparent increase in oxygen consumption is probably due to the oxidation of sulphide to thiosulphate. At higher concentrations, oxygen consumption decreases, perhaps because of the inhibitory effect of sulphide on electron transport, and N. norvegicus resorts to anaerobic metabolism as indicated by the accumulation of lactate in the haemolymph and in the tissues.Communicated by L. Hagerman, Helsingør     

Protein synthesis and growth inOctopus vulgaris

Rates of protein synthesis in the whole body and tissues ofOctopus vulgaris collected in September 1985 and 1986 from the Bay of Naples were measured following a flooding dose injection of³H phenylalanine. There were stable phenylalanine free pool-specific radioactivities and linear incorporation of radiolabel into arm-tip protein from 10 to 30 min after the injection. In starved individuals there were no significant differences between the fractional rates of protein synthesis of the following tissues: ventricle, brain, branchial heart, arm tip, gill, stomach, arm, renal appendage and mantle. The mean value (± SE) for all the tissues was 3.02 ± 0.17% d^–1. In individuals fed varying amounts of crab, resulting in differing growth rates, there was a linear increase in fractional rates of whole-body protein synthesis with growth rate. A standard 148 g octopus growing at 3.0% d^–1 synthesised 0.54 g of protein, with 0.43 g of this protein retained as growth. The proportion of the total protein synthesis which was retained as growth increased with increasing growth rate; at a maximum growth rate of 6% d^–1, over 90% of the protein synthesised was retained as growth. The ventricle, arm tip, gill, arm and mantle also showed similar patterns of a linear increase in fractional rates of protein synthesis with increased growth rates. The RNA concentrations in the whole body and tissues increased with increasing growth rates, but the major change was an increase in the efficiency of translation. It is concluded that rapid growth rates inO. vulgaris are brought about by high rates of protein synthesis and high efficiencies of retention of synthesised protein and, therefore, little protein degradation.     

Effects of the toxic dinoflagellate <Emphasis Type="Italic">Gymnodinium catenatum</Emphasis> on uptake and fate of paralytic shellfish poisons in the Pacific giant lions-paw scallop <Emphasis Type="Italic">Nodipecten subnodosus</Emphasis>

Juvenile Pacific giant lions-paw scallops Nodipecten subnodosus were fed the toxic dinoflagellate Gymnodinium catenatum, a producer of paralytic shellfish poison (PSP), supplied with Isochrysis galbana (a nontoxic microalgae). Short-term (<24 h) experiments were performed to determine clearance and ingestion rates of G. catenatum. Kinetics of PSP was examined in longer-term experiments (>2 days). At high food concentrations, juvenile scallops showed production of pseudofeces, partial shell valve closure, and reduction in feeding. According to HPLC analysis, the only toxins present in the dinoflagellate G. catenatum and in the scallops were the gonyautoxins (GTXs), except in the labial palps and digestive gland, where trace amounts of saxitoxin (STX) were present in scallops. These tissues could play an important role in toxin biotransformation. The ranking of toxin concentration in tissues was: digestive gland > labial palps > intestine > gills > mantle > adductor muscle, where the total contribution of viscera was more than 80% of the total toxin body burden. Juvenile scallops exhibited no apparent detrimental physiological responses during the long-term feeding experiment. The dinoflagellate may contribute nutrients to the scallop, in addition to the microalgae I. galbana. The dinoflagellate may enhance cell uptake and byssus production. Once PSP accumulated during the first 12 days, it was slowly eliminated. The limited capacity for accumulating toxins in the adductor muscle favors domestic marketing of scallops.     

Field depuration and biotransformation of paralytic shellfish toxins in scallop<Emphasis Type="Italic"> Chlamys nobilis</Emphasis> and green-lipped mussel<Emphasis Type="Italic"> Perna viridis</Emphasis>

Under laboratory conditions, the scallop Chlamys nobilis and the mussel Perna viridis were exposed to N-sulfocarbamoyl toxins (C2 toxin), a paralytic shellfish toxin (PST), by feeding a local toxic strain of the dinoflagellate Alexandrium tamarense (ATDP) that produced C2 toxin exclusively. The bivalves were subsequently depurated in the field, and their depuration kinetics, biotransformation and toxin distribution were quantified. Depuration was characterized by a rapid loss within the first day, followed by a secondary slower loss of toxins. In the fast depuration phase, scallops detoxified PSTs more quickly than the mussels (depuration rate constants for scallops and mussels were 1.16 day^–1 and 0.87 day^–1, respectively). In contrast, the mussels detoxified PSTs more quickly than the scallops in the slow depuration phase, and the calculated depuration rate constants (mean+SE) from day 2 to day 13 were 0.063+0.009 day^–1 and 0.040+0.019 day^–1 for mussels and scallops, respectively. The differences in the appearances of gonyautoxins, GTX2 and GTX3, and their decarbamoyl derivatives, dcGTX2, dcGTX3 and GTX5, which are all derivatives of C2 toxin, indicated active and species-specific biotransformation of the algal toxins in the two bivalves. In both species of bivalves, the non-viscera tissue contained fewer toxins and lower concentrations than the viscera-containing tissue compartment. In scallops, very little toxin was distributed in the adductor muscle. In mussels, most of the PSTs were found in the digestive gland with significant transport of toxins into the digestive gland from other tissues during the course of depuration. The toxin profiles of scallops and mussels differed from each other and from that of the toxic algae fed. A significant fraction of GTX5 was detected in the mussels but not in the scallops. Our study demonstrates a species specificity in the depuration kinetics, biotransformation and tissue distribution of PSTs among different bivalves.Communicated by T. Ikeda, Hakodate     

Patterns of trace-metal distribution in tissues of Pleoticus muelleri (Crustacea: Decapoda: Solenoceridae)

Trace-metal distribution in tissues of the shrimp Pleoticus muelleri Bate from the Patagonian region, Argentina, was related to sex, size and physiological condition. Concentrations of cadmium, copper, manganese and zinc were determined in the digestive gland, male reproductive system and muscle of adult specimens. Significant (p<0.01) sex-dependent differences in mean metal concentrations (g g^-1 wet wt) were found in the following tissues of males and females, respectively: digestive gland, Cu, 82.9 and 30.8; Zn, 32.5 and 44.9; reproductive system, Zn, 12.8 and 38.6; Mn, 1.7 and 3.1; Cd, 0.29 and 0.58. Metal levels of muscle showed no significant differences between sexes. Cadmium was not detected in muscle, suggesting that its incorporation into this tissue was strictly regulated. Metal concentrations in the male reproductive system were in general weakly correlated (0.001<p<0.05) with either body size or the concentration of metals in the digestive gland. The developing ovary incorporated substantial amounts of zinc, while cadmium and copper levels decreased at proportional rates (p<0.001). The patterns of the variations in the concentrations of these metals in the ovary strongly suggested that a regulatory mechanism operated throughout oogenesis. Manganese was not involved in this presumptive mechanism. The results of this study are discussed in terms of concentrations and in absolute amounts for standardized individuals.     

Influence of temperature and salinity on the uptake,distribution and depuration of mercury,cadmium and lead by the black-lip oyster Saccostrea echinata

Saccostrea echinata (Quoy and Gaimard) were exposed to 10 g 1^-1 of either mercury, cadmium or lead at 30 °C, 36S; 30 °C, 20S; 20°C, 36S and 20°C, 20S for 30 d and were then transferred to clean seawater for a further 30 d to depurate. Specimens were removed at regular intervals during the exposure and depuration periods, dissected into gills, mantle, visceral mass and adductor, and analysed for the appropriate metal by atomic absorption spectroscopy. Mercury was concentrated more than the other metals in all tissues under all conditions. Cadmium uptake was greater than lead in all tissue in the high-temperature experiments, whereas both metals were concentrated to similar extents at low temperature. The gill tissue generally accumulated the greatest amount of all 3 metals, whilst the adductor concentrated the least amount. At both salinities, mercury and cadmium accumulation by all tissues was significantly greater at the higher temperature whereas lead uptake was only marginally increased. The accumulation rates of mercury at high temperature were significantly greater in all tissues at low compared with high salinity, whereas at low temperature, differences were not significant. Accumulation rates of cadmium and lead in the majority of tissues examined were significantly greater in lowsalinity water at both temperatures. In general, lead was lost the most rapidly from oyster tissues, followed by mercury and then cadmium. The residence times for mercury and cadmium differed significantly between tissues, with the gills showing the highest turnover rate. In contrast, residence times for lead were similar between tissues. Losses of all 3 metals from oyster tissues were not obviously influenced by temperature and only mercury losses differed significantly between salinities.     

Dynamics and physiology of saxitoxin production by the dinoflagellatesAlexandrium spp.

Toxin content (fmol cell^–1) and a suite of elemental and macromolecular variables were measured in batch cultures of the dinoflagellatesAlexandrium fundyense, A. tamarense andAlexandrium sp. from the southern New England region, USA. A different perspective was provided by semicontinuous cultures which revealed sustained, steady-state physiological adaptations by cells to N and P limitation. Two types of variability were investigated. In batch culture, changes in nutrient availability with time caused growth stage variability in toxin content, which often peaked in mid-exponential growth. A second type of variability that could be superimposed on growth stage differences is best exemplified by the high toxin content of cells grown at suboptimal temperatures. Calculations of the net rate of toxin production (R _tox ; fmol cell^–1 d^–1) for these different culture treatments and modes made it possible to separate the dynamics of toxin production from cell division. Over a wide range of growth rates, cells produced toxin at rates approximating those needed to replace losses to daughter cells during division. The exception to this direct proportionality was with P limitation, which was associated with a dramatic increase in the rate of toxin production as cells stopped dividing due to nutrient limitation in batch culture. Growth stage variability in batch culture thus reflects small imbalances (generally within a factor of two) between the specific rates of toxin production and cell division. N limitation and CO₂ depletion both affect pathways involved in toxin synthesis before those needed for cell division; P limitation does the opposite. The patterns of toxin accumulation were the same as for major cellular metabolites or elemental pools. The highest rates of toxin production appear to result from an increased availability of arginine (Arg) within the cell, due to either a lack of competition for this amino acid from pathways involved in cell division or to increased de novo synthesis. There were no significant changes in toxin content with either acclimated growth at elevated salinity, or with short term increases or decreases of salinity. These results demonstrate that toxin production is a complex process which, under some conditions, is closely coupled to growth rate; under other conditions, these processes are completely uncoupled. Explanations for the observed variability probably relate to pool sizes of important metabolites and to the differential response of key biochemical reactions to these pool sizes and to environmental conditions.     

Kinetic sodium analyses in the crab Carcinus mediterraneus

Kinetic analyses were made of the stable sodium and radioactive sodium (²²Na) in some tissues of the crab Carcinus mediterraneus CSRN. Fast ²²Na outflux constant is increased in gills, hepatopancreas, hemolymph, digestive tract and muscles. In the reproductive organs the values of the sodium outflux constant are lower. The values of the fast outflux rates (t=20°C) of sodium amount, in the whole animal, to 22.5 M Na/g/h, in the hemolymph, to 69.3 M Na/g/h; the slow outflux rates have much lower values (0.02 to 0.13 M Na/g/h). Moulting crabs show a rate of sodium outflux 6 times greater than that of intermoulting individuals.     

Metallothioneins and lysosomes in metal toxicity and accumulation in marine mussels: the effect of cadmium in the presence and absence of phenanthrene

It has been confirmed that metallothioneins play an important role in the accumulation of cadmium (Cd) in the digestive gland cells of mussels (Mytilus galloprovincialis Lam.). The content of Cd in the tissue of mussels exposed for 9 d to the metal (estimated dosage of 180 g Cd mussel^-1 d^-1) was 66.2 ppm. This value is about the same as the metal content found in the digestive gland of Cd-exposed mussels kept in clean water for a recovery period of 28 d. At the end of the recovery period, however, the Cd bound to thionein had increased by approximately 250%. Our data demonstrate that the stability of lysosomes, a biological parameter adopted as a cellular stress index, is extremely low in mussels exposed to Cd for 9 d, but returns to control values in the digestive gland cells of mussels allowed to recover for 28 d in uncontaminated sea water. At this point most of the Cd present in the cytosol is bound to thionein. These data demonstrate the importance of metallothionein induction in the reduction of the cytotoxic effects exerted by high levels of Cd accumulation. The results of tests designed to clarify the reasons for the long biological half-life of Cd demonstrated that, in the digestive gland of mussels, the lysosomes are not able to eliminate Cd either bound to insoluble thionein polymers or to lipid peroxidation products such as lysosomal lipofuscin, both of which are apparently involved in the elimination of copper. The absence of these two mechanisms of metal sequestration and elimination via excretion of residual bodies (tertiary lysosomes) is in agreement with the persistence of cadmium in the digestive gland of mussels. Finally, the results also demonstrate that simultaneous exposure of mussels to Cd and phenanthrene, an established lysosomal membrane destabilizer, did not significantly alter the accumulation of Cd or the kinetics of the metal in mussels.     

The distribution of zinc in the oyster Ostrea edulis and its relation to enzymic activity and to other metals

The role of zinc in the oyster Ostrea edulis Linnaeus has been studied in its relation to the zinc-dependent enzymes present and in relation to the copper, calcium, magnesium, sodium, potassium and phosphate contents. Only carbonic anhydrase, alkaline phosphatase, carboxypeptidase A and malic dehydrogenase zinc metalloenzyme activities could be detected. -D-mannosidase, a zinc-dependent enzyme hitherto not reported for the oyster, was also detected. After tissue dissection into muscle, palps, gills, mantle and digestive mass and subcellular fractionation of these tissues, analysis indicated that no single tissue concentrates zinc or the zinc-dependent enzymes. The total amount of zinc found is far in excess of the amount of zinc contributed by the zinc-dependent enzymes, but the amount of non-dialysable zinc is of the same order of magnitude. It is suggested that this apparent excess of dialysable zinc is a consequence of the high levels of calcium found in the tissues, demonstrating a competition between calcium and zinc in their uptake, as is well documented in many other phyla.     

Allelopathic effect of Cylindrospermopsis raciborskii extracts on the germination and growth of several plant species

Cylindrospermopsis raciborskii is an invasive cyanobacterium and a potential producer of the alkaloid toxin cylindrospermopsin (CYN). Extracts of two strains of C. raciborskii were tested for their effects on the germination and growth of Lactuca sativa, Phaseolus vulgaris, Pisum sativum and Solanum lycopersicum. Germination was not significantly inhibited for any of the plant species tested, but growth was affected, depending on the species. Root and stem growth in L. sativa was generally stimulated by both strains. Ph. vulgaris root growth was stimulated by both strains but no effect was visible in stem growth. S. lycopersicum root growth was inhibited by both strains and stem growth was inhibited only by the CYN strain. P. sativum root growth was also inhibited by both strains but stem growth was stimulated. CYN accumulation was also differential with toxin transfer to the stem. Ph. vulgaris accumulated the highest CYN concentration. This study suggests that plants behave differently in their response to this toxin and that roots and stems also show different abilities to react and accumulate the toxin. Knowledge of the impact of CYN- and non-CYN-producing cyanobacteria in different plant species and translocation of the toxin to different plant parts is essential for the avoidance of human as well as environmental health hazards.