Regulation of body dimethylsulfoniopropionate (DMSP) content by the copepod Temora longicornis: a test of four mechanisms

Copepods contain dimethylsulfoniopropionate (DMSP) in their tissues in addition to the DMSP in their gut contents and therefore constitute an additional pool of DMSP in the ocean. In the estuarine copepod Temora longicornis Müller, this dynamic pool of DMSP is reduced when external salinity decreases. In the present study the mechanism(s) used by T. longicornis to adjust its DMSP content were examined. Four possible mechanisms were tested in experiments conducted between July 1997 and May 1999: (1) DMSP cleavage, (2) demethylation, (3) conversion to methionine, and (4) release from the body. Tissue extracts of T. longicornis did not exhibit the ability to cleave or demethylate DMSP. In incubation experiments, when external salinity decreased, T. longicornis individuals reduced their DMSP content without increasing their methionine content. The total amount of DMSP in the incubation vials was conserved regardless of salinity. The copepods retained most of the DMSP in their tissues in 30‰ water, but when salinity was reduced to 20‰, the copepods released 41 to 55% of their body DMSP to the surrounding medium; 89 to 91% of the total DMSP was recovered. This suggests that estuarine copepods represent a reservoir of DMSP, which is released as dissolved DMSP upon salinity decline. Therefore, while osmoregulation by these copepods may have little impact on the chemical properties of DMSP, it will affect the partitioning of DMSP between particulate and dissolved phases in the water column. Received: 20 August 1999 / Accepted: 28 March 2000     

Production of DMSP and DMS during a mesocosm study of an Emiliania huxleyi bloom: influence of bacteria and Calanus finmarchicus grazing

We investigated the influence of bacteria and metazooplankton on the production of dimethylsulfoniopropionate (DMSP) and dimethylsulfide (DMS) during blooms of Emiliania huxleyi (Lohmann) Hay and Mohler in seawater mesocosms. The phytoplankton succession was marked by the rapid collapse of an initial Skeletonema costatum (Greville) Cleve bloom followed by a small E. huxleyi bloom. The collapse of the diatom bloom was accompanied by an increase in concentrations of dissolved DMSP (DMSP_d) and bacterial abundance and activity (as determined by the thymidine incorporation technique). The increase in bacterial activity was followed by a rapid decrease in DMSP_d concentrations which remained low for the rest of the experiment, even during the subsequent collapse of the E. huxleyi blooms. The absence of DMSP_d and DMS peaks during the declining phase of the E. huxleyi blooms was attributed to the high bacterial activity prevailing at that time. The influence of metazooplankton grazing on DMSP and DMS production was investigated by adding moderate (24 mg dry weight m^-3) and high (520 mg dry weight m^-3) concentrations of Copepodite Stage V and adults of Calanus finmarchicus to two of four filtered (200 m mesh net) enclosures during the E. huxleyi blooms. The addition of C. finmarchicus, even in high concentrations, had no apparent effect on the dynamics of E. huxleyi, suggesting that the copepods were not grazing significantly on nanophytoplankton. The addition of copepods in high concentrations favored an accumulation of chlorophyll a and particulate DMSP. These results suggest that copepods were preying on the herbivorous microzooplankton which, in turn, was controlling the biomass of nanophytoplankton. DMS production was also enhanced in the enclosure with maximum metazooplankton biomass, suggesting that the grazing of C. finmarchicus on microzooplankton containing DMSP may contribute to DMS production. These results provide strong support to the emerging idea that bacteria and metazooplankton grazing play a dominant role in determining the timing and magnitude of DMS pulses following phytoplankton blooms.     

Organic solute accumulation in osmotically-stressed Enteromorpha intestinalis

The role of organic solutes in the osmotic adjustment processes of the marine macroalga Enteromorpha intestinalis (L.) Link was investigated in 1986, using fresh samples collected from mid-shore rock pools at Tayport, Fife, Scotland. Natural-abundance ¹³C nuclear magnetic-resonance spectroscopy revealed -dimethylsulphoniopropionate (DMSP) to be the only major low molecular weight organic osmolyte present. However, on transfer to a hypersaline medium (300% sea water; 100%=35 S), tissue sucrose and proline levels increased markedly, while DMSP remained constant. Recovery of optimal photosynthetic activity and increases in inorganic ion levels occurred over a similar time scale to the changes in sucrose and proline (within 48 h), indicating that these two organic solutes are involved in hyperosmotic adjustment in E. intestinalis while DMSP is not. Freshly-collected plants transferred to 300% sea water medium in the dark showed no significant increases in organic osmolytes. In contrast, starch-enrichment (16 d continuous illumination) led to enhanced synthesis of sucrose and proline in the light and in darkness, but tissue DMSP levels showed no variation throughout. These observations suggest that DMSP is not involved in short-term osmoacclimation in E. intestinalis.     

Growth pattern andβ-dimethylsulphoniopropionate (DMSP) content of green macroalgae at different irradiances

Growth rates and intracellular-dimethylsulphoniopropionate (DMSP) concentrations of five green algal species collected from different geographic regions in 1986 and 1989 were determined under four photon flux rates. InUlothrix implexa, U. subflaccida andAcrosiphonia arcta from Antarctica, growth was light-saturated at lower irradiances than in temperateUlva rigida from Southern Chile andBlidingia minima from Germany. The DMSP content ofUlothrix implexa, A. arcta andUlva rigida was directly correlated with the light factor: with increasing irradiance, algal DMSP level increased. In contrast, inUlothrix subflaccida andB. minima DMSP concentrations gradually decreased up to a photon flux rate of 30µmol m^–2 s^–1, then increased markedly under the highest photon flux rate tested. In non-growing, dark-incubatedA. arcta DMSP content was reduced by 35%, while the DMSP pool of all other species remained unchanged, at the level of pre-culture conditions. Under full darkness all plants exhibited a significantly higher DMSP concentration compared with algae grown at low photon flux rates of 2 to 30µmol m^–2 s^–1. These data show a correlation between growth pattern and DMSP biosynthesis, and may point to a species-specific minimum amount of light energy necessary for DMSP accumulation.Contribution no. 302 of the Alfred Wegener Institute of Polar and Marine Research     

Uptake of cadmium by the eelgrassZostera marina

The uptake of cadmium by the shoots ofZostera marina L. (eelgrass) was examined in the laboratory. Experiments were carried out in experimental chambers which allowed the separation of the leaves from the root-rhizome portions of intact shoots. Cadmium uptake by the root-rhizome portions over 24 h was directly related to substrate cadmium concentration (1.0, 5.0 and 10.0 g Cd ml^-1) and varied from 6.5 to 30.0 g Cd g^-1. Cadmium uptake by the root-rhizomes and the leaves in a substrate concentration of 1 g Cd ml^-1 was also related to exposure times (24, 48 and 72 h). Maximum uptake by the root-rhizomes and the leaves was observed after 72 h at a substrate concentration of 1 g Cd ml^-1 and was equivalent to 48 and 94 g Cd g^-1, respectively. Translocation of cadmium from the leaves to the root-rhizomes was observed after 24 h, and at the end of 72 h was equivalent to 27% of the total leaf uptake. No cadmium movement from the root-rhizomes to the leaves was detected.     

Levels of intracellular free amino acids used for salinity tolerance by oysters (Crassostrea virginica) are altered by protozoan (Perkinsus marinus) parasitism

Free amino acid (FAA) levels were measured from May through October 1991 in gill tissues of two groups of juvenile oysters (Crassostrea virginica Gmelin), one transferred from a low salinity field site (8) to a field site of high salinity (20) and high Perkinsus marinus (Mackin, Owen, and Collier) prevalence, the other kept at the low salinity field site. Within 24 h, glycine levels in the oysters transferred to high salinity increased 8-fold, taurine concentrations doubled and the total FAA pool rose from 150 mol g^–1 dry wt to 400 mol g^–1 dry wt. Taurine levels reached a plateau within 20 d after transfer to high salinity and remained at that level until P. marinus infections were detected 85 d after transfer. Taurine and glycine levels declined by 40% in the high salinity population as infection intensity increased between 70 and 105 d. Total FAA declined by approximately 33% over this period. The oysters kept at low salinity were not infected and continued to grow while the infected high salinity oysters showed no increase in shell length after Day 85. FAA levels in the low salinity group remained relatively constant throughout the experiment except for an initial rise triggered by an increase in ambient salinity from 8 to 12. The results suggest that salinity tolerance mechanisms in C. virginica may be impaired by P. marinus infection.     

Eulerian sampling of zooplankton from Marseilles' Vieux Port over a 24 h period

A 24 h continuous study of hydrological parameters and zooplanktonic populations was carried out in Marseilles' Vieux-Port, a semi-closed area supposed to be a fairly homogeneous body of water. Sampling was performed in spring 1980 with a pump system. Samples were taken every 30 min over a period of 24 h, during which time only slight temperature variations were recorded (min.=14.5°C, max.=15.1°C); salinities were high and fairly stable (min.=37.49, max.=37.63%.). It can thus be assumed that the water was homogeneous. Concentrations of suspended particulate matter were high and fairly stable (x = 11.8 mg 1^-1). In most samples, nitrate levels were fairly low (x = 0.84 g-at 1^-1); conversely, phosphate contents fluctuated greatly (min.=0.24 g-at 1^-1, max.=2.24 g-at 1^-1). As a result of these fluctuating nutrient-salt values, the N:P ratios were often unbalanced (between 0.5 and 2.0). Detergent values were low (often less than 0.2 ppm). Zooplanktonic populations were made up of a few eurybiotic species, the most thriving of which were the cladoceran Podon polyphemoides and the copepod Acartia clausi. These two species perform nychthemeral migrations towards the surface: P. polyphemoides during the day and A. clausi at night. As a result of these opposite patterns of migrations, the planktonic biomass remained fairly stable near the surface. There were also some meroplanktonic species (especially cirripede larvae) and a small number of other copepods present. This study revealed a regular hydrological and faunal cycle with a return to initial conditions at the end of the 24 h period.     

Sodium-dependent amino acid transport in the chlorophyte Platymonas subcordiformis

Sodium-dependent transport of alanine and serine by Platymonas subcordiformis was demonstrated by evaluating the kinetics of entry of these substrates over a range of concentrations of ambient sodium. The reciprocal of the concentration at which entry rate was half maximal (K _t ) was linearly related to ambient sodium concentration; maximum entry (V _max) was not affected. Entry of labeled amino acids as measured by determining radioactivity in the medium was close to but slightly less than rates of net entry as determined by high-performance liquid chromatography. This difference presumably reflects extrusion of labeled carbon by the cells in a form not detectable by the chromatographic techniques employed. The coupling coefficient (sodium ions per amino acid molecule) for entry of alanine and serine was evaluated and determined to be 2. Methylaminoisobutyric acid was not taken up by P. subcordiformis and had no inhibitory effect on uptake of alanine or serine. B-2-aminobicycloheptane-2-carboxylic acid competitively inhibited uptake of both substrates. The internal sodium concentration of P. subcordiformis was measured, and the maximum gradient energetically favorable for amino acid transport was calculated from data in the present work and drawn from the literature. Assuming that uptake of amino acids is strictly sodium-dependent, an amino acid concentration gradient of the order of 10⁶ (cell:medium) can be achieved. This concentration differential permits net uptake of amino acids by P. subcordiformis from ambient amino acid levels in the nanomolar range.     

Effect of ionizing radiation on haemoglobin of marine lamellibranchs

The effect of ionizing radiation on the iron-linked protein (haemoglobin) of the marine lamellibranchs Anadara granosa (Linn.) and Cardita antiquata (Lam.) from Bombay waters, India is discussed. Purified haemoglobin solutions were exposed to a ⁶⁰Cobalt source delivering a dose of about 4,600 rad/min, at the sample irradiation point. Radiation damage or degradation was measured spectrophometrically by studying changes in the absorption spectra following irradiation in the presence and absence of oxygen. Exposure to ionizing radiation in general caused a decrease in absorption of both haemoglobins, irrespective of location, viz extra-versus intracellular, at Soret (412 m), (540 to 42 m) and (574 to 76 m) peaks, and an increase in absorption at 510 and 630 m. Upon exposure to a higher dose, O₂Hb of C. antiquata showed an increase in absorption at the protein peak (280 m); exposure to a lower dose, however, resulted in decreased absorption. Furthermore, the changes following irradiation were dependent upon the initial state of the pigment. Oxyhaemoglobin, when exposed to radiation, oxidized to hemiglobin, and hemiglobin reduced to oxyhaemoglobin. The extracellular haemoglobin of high molecular weight (3x10⁶) of the false cockle C. antiquata was found to be extremely radio-resistant, whereas intracellular haemoglobin of low molecular weight (74,000) of the arcid clam A. granosa was highly radiosensitive, since it could not be exposed to doses exceeding 18,000 r.     

The composition and biosynthesis of lipids in Thysanoessa raschi from the Clyde Estuary,Scotland

The lipid composition and biosynthesising activity of Thysanoessa raschi collected from the Clyde Estuary, Scotland, in May 1981 were examined. Triacylglycerols were the major lipid class present, although 16.7% of the total lipid were wax esters in which phytol was the dominant fatty alcohol. The thoracic contents (hepatopancreas) of the krill were capable of biosynthesising lipids in vitro from various labelled substrates. Radioactivity from [1-¹⁴C] palmitic acid was incorporated into lipids in the order phospholipids>triacylglycerols>wax esters; the bulk of the radioactivity was present in all cases in the fatty acyl moieties of the lipids. [U-¹⁴C] glucose labelled lipids in the order phospholipids>triacylglycerols>free fatty acids> was esters; in the first two lipids the radioactivity was mainly in the glycerol moieties, whereas in was esters it was solely in the fatty acyl moieties. The extent of labelling of these lipids from [U-¹⁴C] alanine was less than that from [U-¹⁴C] glucose, but the pattern of labelling was generally similar. More than 90% of the radioactivity incorporated into total lipid from ³H₂O was present in free fatty acids from which it was calculated that the hepatopancreas of T. raschi can synthesise 2.5 g of fatty acid per hour at 15°C. This value is approximately three times lower than that previously determined for T. inermis from Balsfjorden, northern Norway. The results are discussed in terms of the sources of the dietary lipids of krill and the role of endogenous biosynthesis in contributing to its lipid reserves.     

Naturally high levels of zinc and metallothionein in liver of several species of the squirrelfish family from Queensland,Australia

Recently, we found that liver of squirrelfish (Holocentrus rufus) from Bermuda contains remarkably high concentrations of metallothionein (MT) and Zn. To clarify the nature and cause of the Zn accumulation, four Holocentridae species from three genera (Neoniphon sammara, Sargocentron spiniferum, Myripristis violacea, and Myripristis murdjan) were collected from a totally different area, the Great Barrier Reef, Australia. The average hepatic Zn levels in these species ranged from 67 g g^-1 (S. spiniferum) to 1000 g g^-1 (N. sammara). The intraspecies variation in the hepatic Zn level was pronounced. In both N. sammara and M. murdjan, there was a 48-fold difference between the lowest and the highest Zn concentration. Zn accumulation was closely correlated to a dramatically elevated concentration of MT in liver. In N. sammara, which possessed the highest level of MT among the investigated species, the mean hepatic MT concentration was 19 500 g g^-1. Subcellular fractionation of the liver showed that MT was responsible for the binding of up to 74% of the hepatic Zn. The concentrations of MT and Zn in gills were low, suggesting that Zn storage in fish within this family is tissue-specific. The results further indicate that the high concentrations of MT and Zn in Holocentridae liver are linked to normal physiological processes. We suggest that the Holocentridae family may be a unique model for future studies of Zn metabolism and the function of MT.     

HPLC analysis of nonprotein thiols in planktonic diatoms: pool size,redox state and response to copper and cadmium exposure

A sensitive method was developed to analyze low molecular weight thiols involved in metal homeostasis and detoxification in phytoplankton. The aims of this study were to (1) separate and measure all relevant thiols in a single HPLC run, (2) measure redox states of the thiols and (3) identify specific responses of thiols (pools, redox) to heavy metals by testing diatoms with different metal tolerances (Ditylum brightwellii, Phaeodactylum tricornutum, Skeletonema costatum andThalassiosira pseudonana). Copper or cadmium were dosed at a maximum tolerable, species-dependent level, to exponential phase cells growing in artificial medium (14 salinity). Loss of cell viability was monitored by the decrease of fluorescein fluorescence after a 24-h metal exposure. Thiols in extracts of exposed cells and controls were labeled with monobromobimane. Picomoles of cysteine, glutathione, -glutamylcysteine and phytochelatins (PCs) were detected and separated by reversed-phase high performance liquid chromatography. Cysteine increased in all species after metal exposure. The species with the largest glutathione pools in the control (P. tricornutum) synthesized the largest PC pools upon metal exposure, however, at a 40% loss of glutathione. A considerable increase of glutathione was observed inD. brightwellii upon metal exposure. However, it produced little PC (and only with Cu). In controls ( 3 pM Cu²⁺), PC₂ was detectable inS. costatum, P. tricornutum andT. pseudonana. Oxidized thiol fractions were recovered by the reductants DTT and TCEP, both performing identically. Compared to the other thiols, cysteine had low redox ratios. InD. brightwellii glutathione and PC redox ratios were lower than inP. tricornutum, S. costatum andT. pseudonana. It was expected that Cu-induced oxidative stress would decrease the thiol redox ratios, however, this was not observed.This is Communication No. 2172 of NIOO.     

Growth increments in the shell of the living brachiopod Terebratalia transversa

Scanning electron micrographs revealed growth increments in the primary shell layer of the extant terebratulid brachiopod Terebratalia transversa collected from Anacortes, Washington, USA, during the summers of 1982–1984. The increments extend into the secondary shell layer, but only as poorly-defined continuations. Growth increments narrow and widen cyclically, producing patterns similar to tidally-induced, fortnightly growth patterns in bivalve molluscs. Asymmetric increments or doublets that consist of paired wide and narrow bands present additional evidence that the growth pattern is produced by the interference of tidal (24 h 50 min) and solar (24 h) daily environmental influences on growth, as is the case in bivalves. Growth increments are relatively well-defined in specimens from tidal habitats and are poorly-defined in subtidal specimens. Specimens grown in laboratory aquaria and subjected to simulated tidal emersions and submersions had well-defined increments, whereas those that were continuously submerged in aquaria had poorly-defined increments, paralleling the distinctness in growth increments produced at various depths in the natural habitat. Production of growth increments in T. transversa may be related to valve movement and changes in mantle-cavity pH. When the valves are closed, pH drops and, when they are open, pH rises. Highperformance liquid chromatography (HPLC) of mantle tissue and fluid failed to detect any single organic acid that could have been responsible for the pH changes, confirming that intermediate metabolism in T. transversa is very low compared to that of bivalve molluscs. Calculations indicate that cycles in intermediate metabolism may nevertheless be responsible for cycles in growth-increment production and shell dissolution.Please address all correspondence to Dr. G.D. Rosenberg in Indianapolis     

Avoiding predation: alarm responses of Caribbean sea urchins to simulated predation on conspecific and heterospecific sea urchins

Alarm responses to the extracts of conspecifics and hetero-specifics were measured for the Caribbean sea urchins Echinometra viridis, E. lucunter, Lytechinus variegatus, L. williamsi, Tripneustes ventricosus, Diadema antillarum, and Eucidaris tribuloides collected along the Caribbean coast (9°3314N; 78°5523W) during October 1984 and July–December 1985. Responses to seawater and extracts of the gnathostomate echinoid Clypeaster sybdepressus were used as controls. The intensity of the response resulting from exposure to sea-urchin extracts was measured by: (1) the percentage of individuals that responded by moving away from the extract and/or towards shelter, and (2) the mean distance moved. Echinometra viridis, E. lucunter, and L. williamsi responded to sea-urchin extracts by moving towards nearby shelter sites. The distance that individuals of each species moved in the first minute following exposure to conspecific extracts was correlated with the distance that species moved from shelter while foraging. L. variegatus and D. antillarum, living in microhabitats not providing protection from predators, responded to extracts of conspecifics and heterospecifics by moving away from the direction of the extract. Eucidaris tribuloides did not exhibit alarm responses to the extracts of con- or hetero-specifics. E. tribuloides secures itself with its stout spines into protected sites within corals. Similarly, L. variegatus living in long, dense seagrass that provided protection from detection by predators, and D. antillarum occupying crevices, showed no alarm responses to extracts of conspecifics. Presumably, in these situations, sea urchins cannot increase their defenses against predation by moving away from an injured neighbor. T. ventricosus showed a weak response to extracts of L. variegatus, but no response to extracts of other species including conspecifics. The reasons for this lack of an alarm response are unclear. For the five species that demonstrated an alarm response to sea-urchin extracts, the intensity of the response varied depending on the type of extract used. L. variegatus, L. williamsi, and D. antillarum responded most strongly to extracts from conspecifics, while Echinometra viridis and E. lucunter responded strongly to extracts from both conspecifics and congeners. The weakest responses were shown to the extracts of T. ventricosus and Eucidaris tribuloides. Habitat overlap, overlap in predators, and phylogenetic relationships did not consistently explain patterns of alarm responses to the extracts of heterospecific sea urchins.     

β-glucanase activity in the seastar Pisaster ochraceus

Specimens of Pisaster ochraceus, an intertidal carnivorous seastar, were collected in northern Puget Sound at Post Point, near Bellingham, Washington, USA, in November 1993, and used in experimentation through September 1994. Ammonium sulfate fractions (60% precipitates) from excised pyloric caeca (digestive glands) possessed -glucanase activity directed toward such -linked substrates as carboxymethylcellulose (CMC), partially-digested cellulose (cellodextrins) and the synthetic substrate p-nitrophenyl--D-glucoside (PNDG). pH-activity and pH-stability maxima were at pH 6 and 5, respectively. No hydrolytic activity was detectable on native cellulose, native chitin or on cationic-modified celluloses such as DEAE-cellulose. Gel filtration (Sephadex G-100) of this 60% precipitate followed by DEAE-Sephadex column chromatography revealed heterogeneity of -glucanase activity, with an indication of up to 11 distinct fractions with variable activity directed toward the substrates indicated above (CMC, cellodextrins and PNDG). These fractions displayed differing ratios of hydrolytic activity on these substrates. None of the isolated fractions displayed activity toward either native cellulose or native chitin. Gut extracts from the edible mussel (Mytilus edulis), a common prey species of P. ochraceus, were shown to possess partially digested cellulose (cellodextrins). These cellodextrins were further degraded by seastar -glucanases in vitro. The digestion of the gut contents of ingested prey provides a possible adaptive explanation for the presence of -glucanase activity in a carnivore.     

Spore germination and early stages of development in Hypnea musciformis (Rhodophyta,Gigartinales)

The early stages of development of the carpospores and tetraspores of Hypnea muscilormis (Wulfen) Lamouroux have been investigated. Both types of spores germinated immediately after liberation. The spores segmented repeatedly into, 2, 3, 4 cells, etc. until a multicellular ball of cells was produced. The germination pattern was thus of the discal type, i.e., Typus discalis mediatus (Inoh, 1947) or the Dumontia-type in the terminology of Chemin (1937). Subsequently, a relatively expansive attachment holdfast was produced from each sporeling. This was followed by the development of up to 4 or more shoot axes arising from the basal disc. These findings are discussed with reference to an earlier study by the senior author on the developmental biology of other species of Hypnea Lamouroux.     

Growth,condition and specific dynamic action in the mussel Mytilus edulis recovering from starvation

Juvenile Mytilus edulis were grown individually in plastic racks in a tidal salt marsh for 72 d in 1984, starved in the laboratory for 130 d, and then fed the alga Isochrysis galbana daily for 64 d. Oxygen consumption was measured at various times during the course of starvation and recovery. The effects of both size (tissue dry weight) and condition (tissue size relative to shell size) on the rate of oxygen consumption during the course of starvation and recovery were analyzed by multiple regression. Weight-specific preprandial rate was inversely correlated with both size and condition. Weight-specific active rate (measured shortly after feeding) was correlated with size but not condition. Relative Scope for Activity was inversely correlated with size and positively correlated with condition. Relative Specific Dynamic Action (RSDA; the integrated physiological and mechanical response to a meal) was initially correlated negatively with size and subsequently positively with condition. Glycogen content was shown to be positively correlated with condition in mussels before starvation and during recovery. During recovery, experimental mussels returned to 90% of their estimated dry weight prior to starvation, and from 53% after starvation. At weekly intervals during recovery, oxygen consumption was measured following a meal until it returned to the preprandial rate. Both pre- and postprandial volumes (l at STP) of oxygen consumed per hour ( O _2pre and O _2post , respectively) increased significantly during recovery. The postprandial rise in oxygen consumption increased significantly from 15% to 23% of O _2pre . At the end of the recovery period, RSDA [( O _2post – O _2pre )/ O _2pre ] was independent of final dry weight, but was significantly correlated with percent dry weight recovered (r ²=0.44; df=10; P<0.02). The increase in RSDA may reflect increased utilization of food and its conversion to soma during recovery from starvation, as distinct from mechanical energy expenditure (feeding activity) following a meal.     

Metal interaction during accumulation by the mussel Mytilus edulis planulatus

Mytilus edulis planulatus (Lamarck) were collected from Howden, South-east Tasmania in autumn 1981. Interaction effects of cadmium, copper and zinc during accumulation by mussels exposed for ten days to all three metals simultaneously were examined in a series of experiments in which each metal was tested at three concentrations. In general, interaction effects were most evident at the highest concentrations tested (20 g l^-1 Cd; 20 g l^-1 Cu; 200 g l^-1 Zn) and led to a reduction in the accumulation of cadmium and an increase in that of copper and zinc. More specifically, high levels of zinc caused a decrease in cadmium uptake and an increase in copper accumulation. The presence of copper resulted in depressed cadmium accumulation while zinc accumulation increased. Cadmium tended to enhance zinc accumulation, but copper accumulation was only affected to any great extent when zinc was also present.     

Oxygen-binding by haemocyanins from an ecological series of amphipod crustaceans

The oxygen-binding properties of haemocyanins (Hc) from three species of gammaridean amphipods, Gammarus locusta (L.) (subtidal), Echinogammarus pirloti (Sexton and Spooner), (intertidal, marine) and E. marinus (Leach) (intertidal, estuarine), one species of hyalid amphipod Hyale nilsonni Rathke (high intertidal, marine) and the talitrid amphipod Orchestia gammarellus (Pallas) (semi-terrestrial) have been studied. All the species were collected from the Firth of Clyde, Scotland, during the spring of 1992. The oxygen-carrying capacity of haemolymph from each species was low, although variable, and was correlated with the low concentration of Hc present. The Hc oxygen-affinity of native gammarid haemolymph was relatively high [partial pressure of oxygen required for half-saturation, P₅₀=4 to 5 torr (0.53 to 0.67 kPa)] at their respective in vivo pH values. At equivalent pH, however, Hc from G. locusta displayed a lower O₂-affinity than either Echinogammarus species. Gammarid Hcs had a large Bohr effect ( log P₅₀/ pH=-1.16 to-1.47). Resuspended Hc isolated from whole H. nilsonni showed similar O₂-binding properties to those of the gammaridean amphipods [P₅₀=6.3 torr (1.44 kPa) at pH=8.0; log P₅₀/pH=-1.20]. Comparable data for haemolymph from O. gammarellus showed that the Hc had a lower affinity for O₂ [P₅₀=14.1 torr (1.87 kPa) at in vivo pH] and exhibited a more moderate Bohr effect ( log P₅₀/ pH=-0.79). To eliminate the possibility that these differences were due to the different haemolymph constituents, each of the Hcs were pelleted and resuspended in physiological saline. The differences noted above persisted, demonstrating that they were due to inherent O₂-binding properties of the Hc molecules themselves. An increase in L-lactate resulted in an increase in Hc oxygen-affinity for both Echinogammarus species but not for O. gammarellus. This study has confirmed that there is a clear difference between Hcs from aquatic and semi-terrestrial amphipod genera. The results lend further support to the hypothesis that the move on to land by amphipod crustaceans is accompanied by a decrease in Hc oxygen-affinity, a decrease in the Bohr effect and a decrease in effector (in this case L-lactate) sensitivity.     

Notes on some factors affecting aggressive behaviour in Carcinus mediterraneus

The relationships between aggressive behaviour and animal density and between aggressive behaviour and metabolic excretion have been investigated in Carcinus mediterraneus Czerniavsky. Increasing the crab density by increasing the number of crabs or reducing the available area revealed that the aggressive interactions perindividual did not increase. The frequency of interactions reached a maximum in several minutes following the introduction of crabs. Individuals isolated between 6 and 24 h were more aggressive than non-isolated ones. The water in which a group of crabs had been held had an inhibitory effect (not abolished by commercial aquarium filters) on aggressiveness of pairs of crabs grater than plain water or water in which single crabs had been kept. No home-group effect was observed. The effect of group water was enhanced by increasing the concentration of crabs in the duration of confinement, and it is probably due to the accumulation of catabolites in the medium. Crabs kept in groups had a greater excretion than isolated crabs and the nitrogen oupput decreased with an increase in the number of crabs or whith a decrease in the size of the tank (with a constant volume of water and number of crabs).