Influence of environmental factors on burrow irrigation and oxygen consumption in the mudflat invertebrate Urechis caupo

We examined burrow irrigation activity by the mudflat worm Urechis caupo in response to suspended food, ambient hypoxia (down to 3.3 kPa PO₂), hydrogen sulfide exposure (up to 100 µmol l^-1), and short-term temperature change (range 10-22°C). In normoxic, nutrient-free water at 14°C, O₂ consumption ( [(M)\dot]O₂ ) \left( {\dot M{\rm O}_2 } \right) was 45 nmol min^-1 g^-1, water flow rate ( [(V)\dot]_W ) \left( {\dot V_{\rm W} } \right) was 27 ml min^-1 (0.66 ml min^-1 g^-1), frequency of peristaltic waves (F_P) was 2.6 contractions min^-1, stroke volume (SV) was 11 ml, and O₂ extraction coefficient (EO₂) was 0.27. Adding suspended food to the burrow water occasionally elicited stereotypical feeding behavior but had no effect on any measured variables during nonfeeding periods. Hypoxia greatly decreased [(M)\dot]O₂ \dot M{\rm O}_2 (75% reduction at 3.3 kPa PO₂) but did not affect [(V)\dot]_W \dot V_{\rm W} , F_P, SV, or EO₂. Sulfide at 50 µmol l^-1 or less had no effect on burrow irrigation activity, whereas 100 µmol l^-1 sulfide decreased [(V)\dot]_W \dot V_{\rm W} by 58% and F_P by 50% but had no effect on SV. Temperature strongly affected [(V)\dot]_W \dot V_{\rm W} (Q₁₀ of 1.9 from 10°C to 22°C). We propose that U. caupo's ability to live in the hypoxic, sulfidic mud of productive mudflat environments, combined with its very efficient mucous net, allows it to process much less water for feeding than other suspension-feeding invertebrates. This, in turn, necessitates an efficient O₂ extraction mechanism, which is provided by the water lung activity of U. caupo's unique hindgut.     

Effect of oxygen concentration on metabolism and locomotory activity of <Emphasis Type="Italic">Moina</Emphasis><Emphasis Type="Italic">micrura</Emphasis> (Cladocera) cultured under hypo- and normoxia

The influence of oxygen concentration on total and basal metabolism, scope of activity, drag force and duration of jerks, time spent swimming and energy cost of locomotion in Moina micrura Hellich females cultured under hypo- and normoxia was investigated. Scope of activity (Q_l) of hemoglobin-rich red individuals (M_a) acclimated to hypoxia depended less upon oxygen concentration than that of non-acclimated, pale individuals (M_na). Within the range 10-0.3 mg O₂ l^-1 Q_l decreased 4.4-fold in M_a and 62.5-fold in M_na. In both M_a and M_na the integral drag force of antenna fell from 0.22ǂ.07 to 0.12ǂ.04 dyn (1 dyn=1·10^-5 N), the duration of jerks increased from 0.06ǂ.01 to 0.1ǂ.02 s in the range from ~2.0 mg O₂ l^-1 to sublethal oxygen concentrations. At 0.7-0.8 mg O₂ l^-1 M_na stopped filtration and increased time spent swimming. In contrast, even under more severe hypoxia (~0.2 mg O₂ l^-1), M_a maintained their filtering activity using energy (up to 80% of total metabolism) achieved due to increased oxygen capacity of the blood. Separating locomotion and feeding functions, M. micrura can spend less energy for swimming and use its energy budget more plastically under changing environmental conditions.     

Temperature effects on the metabolism of larvae of the Antarctic starfish Odontaster validus, using a novel micro-respirometry method

Oxygen consumption of individual larvae of the Antarctic sea-star Odontaster validus was measured during the 50-day period following fertilisation. Values ranged from 0.76 pmol O₂ h^-1 for one specimen at the coeloblastula stage to 77.6 pmol O₂ h^-1 for one bipinnaria larva. At 0°C the mean oxygen consumption rate of an individual larva increased from 10.9 pmol O₂ h^-1 (standard error of the mean, SEM, 0.13) for a gastrula larva, 13 days post-fertilisation, to 25.4 pmol O₂ h^-1 (SEM 3.5) at the bipinnaria stage (50 days post-fertilisation). Gastrulae reared at -0.5°C did not have significantly different oxygen consumption rates between days 13 and 45 post-fertilisation (mean=11.4 pmol O₂ h^-1). Individual metabolic rates were highly variable, covering more than a 40-fold range. At 2°C gastrula oxygen consumption was on average 45% higher (17.35 pmol O₂ h^-1), giving a Q₁₀ temperature effect of 4.4. For bipinnaria, mean oxygen consumption in 2°C larvae (31.4 pmol O₂ h^-1) was not significantly different from that in larvae at -0.5°C, suggesting bipinnaria metabolism may be less sensitive to temperature change than earlier stages. At 2°C the bipinnaria stage was reached at 30-35 days compared with 45-50 days at 0°C, giving a Q₁₀ of 4.5 for temperature effects on development. The method here used a new, highly sensitive micro-respirometry method that is inexpensive and straightforward in design. Individual larvae of O. validus were held in 35- to 50-µl respirometers. These larvae have very low metabolic rates, and published work on such organisms have utilised at least 25 individuals per chamber. The oxygen content of the respirometers was measured using a 25-µl sample injected into a couloximeter. Oxygen consumption rates down to -1 pmol h^-1 can be detected. Under optimum conditions oxygen consumption of a single larva of -4 pmol O₂ h^-1 was measured with an accuracy of ᆨ%. Values of ~15 pmol h^-1 could routinely be measured with this accuracy. This method would allow oxygen consumption to be evaluated in individual field-caught larvae of most marine ectotherms.     

Feeding biology and carbon budget of the sediment-burrowing heart urchin Brissopsis lyrifera (Echinoidea: Spatangoida)

This study of the burrowing heart urchin Brissopsis lyrifera includes measurements on feeding and food selection, ingestion rate, absorption efficiency, ventilation and respiration. B. lyrifera regulated feeding depth, ingestion rate and absorption efficiency in relation to food source. When food was added to the top layer of sediment, B. lyrifera burrowed closer to the surface and ingested mainly enriched surface material, whereas it burrowed deeper and ingested deep-seated sediment in the controls. In non-enriched sediment, the feeding rate was 0.04 g sediment DW h^-1 ind.^-1, and, in macro- and microalga-enriched sediment the feeding rate was 0.06 and 0.08 g sediment DW h^-1 ind.^-1, respectively. Absorption efficiency of TOC was about 43% in non-enriched sediment and in microalga-enriched sediment, but was significantly lower (34%) in macroalga-enriched sediment. Absorption efficiency of N varied between 48% and 55%, and was independent of food source. B. lyrifera feed selectively, enriching the gut TOC content about 2-fold and N content about 2.5-fold. The C/N ratio was therefore lower within the gut compared to the surrounding sediment, while the faecal C/N ratio was almost equal to the surrounding sediment. The faeces were, however, slightly richer in TOC and N compared to the surrounding sediment. For 3-5 cm long B. lyrifera, water current rate varied between 4 and 24 ml water h^-1, with a mean of 11 ml h^-1. Mean respiration rate was 205 µl O₂ h^-1 ind.^-1. The water current rate was not sufficient for B. lyrifera to sustain itself by filter feeding only. However, organic-rich particles from the surface are suggested to be an important contribution to the diet. A carbon budget was calculated for B. lyrifera from measured values of consumption, absorption efficiency and respiration, in order to estimate annual production of B. lyrifera. Compared to literature values, growth was overestimated about tenfold in the budget. A large proportion of the absorbed carbon was suggested to leave the animal as dissolved carbon, through mucus production or through anaerobic pathways, either by the heart urchin or by micro-organisms in the gut.     

Characterization of urease activity in three marine phytoplankton species,<Emphasis Type="Italic"> Aureococcus anophagefferens</Emphasis>,<Emphasis Type="Italic"> Prorocentrum minimum</Emphasis>, and<Emphasis Type="Italic"> Thalassiosira weissflogii</Emphasis>

The availability of different forms of nitrogen in coastal and estuarine waters may be important in determining the abundance and productivity of different phytoplankton species. Although urea has been shown to contribute as much as 50% of the nitrogen for phytoplankton nutrition, relatively little is known of the activity and expression of urease in phytoplankton. Using an in vitro enzyme assay, urease activities were examined in laboratory cultures of three species: Aureococcus anophagefferens Hargraves et Sieburth, Prorocentrum minimum (Pavillard) Schiller, and Thalassiosira weissflogii (Grunow) Fryxell et Hasle. Cultures of P. minimum and T. weissflogii were grown on three nitrogen sources (NO₃^m, NH₄⁺, and urea), while A. anophagefferens was grown only on NO₃^m and urea. Urease was found to be constitutive in all cultures, but activity varied with growth rate and assay temperature for the different cultures. For A. anophagefferens, urease activity varied positively with growth rate regardless of the N source, while for P. minimum, urease activity varied positively with growth rate only for cultures grown on urea and NH₄⁺. In contrast, for T. weissflogii, activity did not vary with growth rate for any of the N sources. For all species, urease activity increased with assay temperature, but with different apparent temperature optima. For A. anophagefferens, in vitro activity increased from near 0-30°C, and remained stable to 50°C, while for P. minimum, increased in vitro activity was noted from near 0-20°C, but constant activity was observed between 20°C and 50°C. For T. weissfloggii, while activity also increased from 0°C to 20°C, subsequent decreases were noted when temperature was elevated above 20°C. Urease activity had a half-saturation constant of 120-165 wg atom N l^у in all three species. On both an hourly and daily basis, urease activity in A. anophagefferens exceeded nitrogen demand for growth. In P. minimum, urease activity on an hourly basis matched the nitrogen demand, but was less than the demand on a daily basis. For T. weissflogii, urease activity was always less than the nitrogen demand. These patterns in urease activity in three different species demonstrate that while apparently constitutive, the regulation of activity was substantially different in the diatom. These differences in the physiological regulation of urease activity, as well as other enzymes, may play a role in their ecological success in different environments.     

Participation of the C-type hemolymph lectin in mineralization of the acorn barnacle Megabalanus rosa

BRA-2, a major C-type lectin in the hemolymph of the acorn barnacle Megabalanus rosa was detected in shell-associated proteins. Immobilized BRA-2 promoted CaCO₃ crystallization as determined by an in vitro pH-drift mineralization assay. In nature, calcification occurs as wound repair in M. rosa after breakage of the shell plate. To observe this phenomenon, we inserted a rubber cube inside the mantle cavity of M. rosa through the basal plate, and kept the organisms in a tank for several weeks. The rubber cube was covered first with brownish-colored material, probably a melanin, and then with CaCO₃ 5 weeks after the insertion. In the CaCO₃-associated proteins, BRA-3, a minor component of M. rosa hemolymph lectins, was present in addition to BRA-2. The presence of both the lectins and their ligands on the hemocyte was also observed. These results suggest that the C-type lectins of the M. rosa hemolymph participate in mineralization as well as defense in these organisms.     

Alarm responses of the green sea urchin, Strongylocentrotus droebachiensis, induced by chemically labelled durophagous predators and simulated acts of predation

The green sea urchin, Strongylocentrotus droebachiensis, exhibited immediate behavioural responses to waterborne chemosensory cues from two durophagous predators, the Atlantic wolffish, Anarhichas lupus, and the edible crab, Cancer pagurus; as well as from crushed conspecifics and crushed blue mussels, Mytilus edulis. The response patterns were dose dependent and diet dependent. Strong responses were elicited by water conditioned by echinivorous wolffish (97.5%), undiluted urchin extract (73%), undiluted mussel extract (52.5%), and by water conditioned by echinivorous crab (45%). In contrast, urchin extract diluted to 1%, mussel extract diluted to 10%, and water conditioned by predators on a mussel diet elicited weak responses (~20%). Infection by the endoparasitic nematode Echinomermella matsi had no significant effect on the response pattern of S. droebachiensis. There was no conclusive evidence of an alarm response to the predators per se, as the weak response to stimuli from non-echinivorous wolffish and crab, as well as from the seastar Asterias rubens, may have been caused by chemical cues transmitted from their mussel diet. The diet-dependent response to predators suggests that active predators were labelled by chemical cues from their echinoid prey. The chemical cue from echinivorous wolffish acted as both an arrestant and as a repellent, whereas the response to other cues was predominantly or exclusively repellent. The response to echinivorous wolffish was quantitatively stronger than, and qualitatively different from, the response to other stimuli, including undiluted urchin extract. The wolffish is apparently being labelled by a latent chemical cue which derives its potency from activation by, or interaction with, substances in the digestive system of the wolffish. We interpret this novel phenomenon as evidence of alarm signal magnification. The induced behavioural modifications demonstrate the green sea urchins' ability to detect chemical cues associated with active durophagous predators, and would therefore seem to have adaptive potential as a predator defence mechanism.     

Carbohydrate metabolism during anoxia and post-anoxia recovery in Chasmagnathus granulata crabs maintained on high-protein or carbohydrate-rich diets

We assessed the effects of anoxia exposure and recovery on glycogen synthesis and mobilization, glucose uptake, and on the enzymes that control carbohydrate metabolism in the hepatopancreas of Chasmagnathus granulata crabs receiving either a carbohydrate-rich (HC) or a high-protein diet (HP). In both dietary groups, anoxia led to a reduction in glucose uptake and in glycogen synthesis, and to an increase in hepatopancreas glycogen mobilization and in hemolymph glucose concentration. During the first 4 h of exposure to anoxia, total glycogen phosphorylase (GPT) and a form activity increased in HP and HC crabs, leading to a decrease in hepatopancreas glycogen concentration. During recovery, HP and HC crabs rapidly restored the hemolymph glucose levels to pre-anoxia concentrations. In HC crabs, incorporation of ¹⁴C from glucose into glycogen increased gradually after 12 h in normoxia, leading to restoration of glycogen concentration. Also during recovery, the ratio of glycogen synthase I (GSI) to glycogen phosphorylase a (GPa) increased in the HC group. In turn, recovering HP crabs had two peaks of glycogen synthesis, related with two peaks in the ratio of GSI to GPa. Consequently, no mobilization of ¹⁴C-glycogen occurred in recovering HP animals. Anoxia in C. granulata induces a marked decrease in the synthesis of carbohydrate reserves that is accompanied by an increase in glycogen mobilization and in circulating glucose levels. During the recovery period, there is an activation of endergonic processes which cause a decrease in hemolymph glucose levels. In C. granulata, glycogen metabolism seems to be controlled by the ratio of the GSI form to the GPa form. In field conditions, theses changes in the metabolic pattern may result from environmental PO₂ availability. In the winter, C. granulata stays in its holes, where environmental PO₂ falls to zero. The carbohydrate or protein content of the diets administrated to the crabs seem to induce different metabolic adjustments during anoxia and recovery.     

Cardiac output during exposure to <Emphasis Type="Italic">Chattonella</Emphasis><Emphasis Type="Italic">marina</Emphasis> and environmental hypoxia in yellowtail (<Emphasis Type="Italic">Seriola</Emphasis><Emphasis Type="Italic">quinqueradiata</Emphasis>)

The experiments were undertaken to measure, for the first time, cardiac output in yellowtail (Seriola quinqueradiata) during exposure to a harmful red tide flagellate (Chattonella marina). The responses were compared with those during exposure to environmental hypoxia to evaluate the significance of the drop of arterial oxygen partial pressure (PaO₂) in the fish-kill mechanisms by C. marina. PaO₂ immediately decreased, whereas heart rate (HR) was maintained until shortly before death during exposure to C. marina. Suffocation developing during the exposure resulted from a decrease in blood oxygen content, but not from lowered blood flow to the tissue. Although exposure to both C. marina and hypoxia immediately decreased PaO₂, arterial oxygen content (CaO₂) and pH (pHa) were significantly lower, whereas HR and cardiac output (&Qdot;) remained significantly higher, for the C. marina-exposed fish than for hypoxia-exposed fish. Although the drop in PaO₂ appears to play a pivotal role in the mechanisms of fish death by C. marina, other physiological response(s) should also be considered.     

Possible involvement of the glycocalyx in the ichthyotoxicity of Chattonella marina (Raphidophyceae): immunological approach using antiserum against cell surface structures of the flagellate

Although the ichthyotoxic mechanism of Chattonella marina is still unknown, several lines of evidence suggest that the reactive oxygen species (ROS), such as superoxide anion (O₂^-), hydrogen peroxide (H₂O₂)_, and hydroxyl radical (·OH), produced by C. marina are involved in the mortality of fish exposed to this flagellate. Recently, we found that the cell-free supernatant prepared from C. marina, which is considered to contain the glycocalyx, showed NADPH-dependent O₂^- generation. In this study, we prepared antiserum against the crude glycocalyx of C. marina. Using indirect immunofluorescence, it was confirmed that the antiserum specifically reacted with C. marina cells. In addition to C. marina, the antiserum also reacted with other raphidophycean flagellates such as Heterosigma akashiwo, Olisthodiscus luteus, and Fibrocapsa japonica, whereas no reactivity was observed against six other flagellate species tested. These results suggest that raphidophycean flagellates have common epitopes recognized by the antiserum. Interestingly, immunohistochemical analysis of paraformaldehyde-fixed gill lamellae from yellowtail exposed to C. marina revealed that the antiserum stained the surface of gill lamellae, while no such staining pattern was observed in control gill lamellae. These results suggest that the glycocalyx may be discharged when C. marina cells are inhaled into the fishes' mouths and then come into contact with the gill surface. Based on the present results, together with our previous findings, we propose that continuous accumulation of the discharged glycocalyx on the gill surface occurs during C. marina exposure, which may be responsible for the ROS-mediated severe gill tissue damage leading to fish death.     

Seasonal variation in ecophysiological patterns in macroalgae from an Arctic fjord. I. Sensitivity of photosynthesis to ultraviolet radiation

Marine macroalgae inhabiting Arctic coastal ecosystems are exposed to pronounced seasonal variations in the radiation regime, including harmful UVB radiation. This study presents the first data on the seasonal changes in the sensitivity of macroalgal photosynthesis towards UV exposure by comparing under-ice, clear-water and turbid-water conditions characteristic for late winter, spring and summer. Various brown (Laminaria saccharina, L. digitata, L. solidungula, Saccorhiza dermatodea, Desmarestia aculeata), red (Palmaria palmata, Devaleraea ramentacea) and one green macroalgal species (Monostroma aff. arcticum) were collected at the same water depth throughout the seasons in the Kongsfjord (Spitsbergen, Svalbard, Norway). Maximum quantum yield (F_v/F_m) and maximum photosynthetic electron transport rates (ETR_max) were determined immediately after collection, after 2 h exposure to artificial UV radiation and after 18 h recovery in dim white light. Photosynthesis of the studied species showed different responses depending on their morpho-functional and physiological characteristics, their life strategies, phenology and depth distribution. Within the genus Laminaria, maximum quantum yield of adult specimens of the deep-water species L. solidungula was most strongly UV sensitive. Adult L. saccharina exhibited a lower UV sensitivity than a 6-month-old specimen. Inhibition of photosynthesis after UV exposure remained at the same level throughout the study period, both in adult L. saccharina and S. dermatodea. However, adult specimens of L. saccharina collected in May showed partial recovery only, whereas photosynthesis of specimens from both species collected later recovered fully. D. aculeata exhibited a remarkable decrease of UV sensitivity during the study period. Photosynthesis of specimens collected under the ice was strongly inhibited by UV, but the degree of inhibition decreased during spring and summer. Concomitantly ETR_max values were low after UV exposure in specimens collected in June, but increased later in the season. P. palmata exhibited a relatively flexible response. Photosynthesis in specimens collected under the ice in June or in turbid water in July/August was relatively strongly inhibited; specimens collected during sunny periods and in clear water in spring showed a much lower degree of photoinhibition after UV exposure. The seasonal pattern of low/high ETR_max values in spring/summer is probably a characteristic of the life strategy of this species. The UV sensitivity of D. ramentacea exhibited a similar seasonal pattern. In M. aff. arcticum, UV sensitivity increased and ETR_max values decreased during the study period, reflecting the life strategy of this annual late winter/spring species. The physiological basis for the seasonal changes in UV sensitivity of photosynthesis is presented in a companion paper (this issue).     

Sulfuric acid in the phaeophyte alga Desmarestia munda deters feeding by the sea urchin Strongylocentrotus droebachiensis

The phaeophyte alga Desmarestia munda (Order Desmarestiales) concentrates sulfuric acid (H₂SO₄) in cell vacuoles. The primary function of the accumulated acid in the alga's life history is unknown. To investigate if sulfuric acid works as a chemical defense, grazing by the sea urchin herbivore Strongylocentrotus droebachiensis on D. munda was compared to grazing on four common non-acidic phaeophyte algae (Alaria marginata, Laminaria saccharina, Costaria costata, and Nereocystis luetkeana) in multiple-choice feeding preference experiments. Both fed and starved urchins discriminated among algae; N. luetkeana and L. saccharina were consumed preferentially to D. munda and A. marginata. Fed urchins consumed a smaller quantity of A. marginata and D. munda than the starved urchins (15-28% less). The lack of preference for D. munda was not correlated with the presence of polyphenolic compounds or with nitrogen concentration. A. marginata had a high polyphenolic concentration (0.8% DM, dry mass) compared to that of the other four algae (0.08-0.24% DM). The nutritional content (C:N ratio and percent nitrogen) of the five algae ranged from 10.8 to 16.2 and from 2.08 to 3.55, respectively, but was not correlated with food choice by the urchins. Sulfuric acid made up nearly 16% DM of D. munda tissue and the pH of homogenized tissues averaged 1.99 pH units. Sulfuric acid, added to prepared food to provide a range of pH values from 1.9 to 6.2 pH units, deterred feeding by urchins at pH values Г.5 pH units. This result provides direct evidence that sulfuric acid at concentrations found in algal tissues deters grazing by S. droebachiensis. Therefore, acid accumulation in D. munda provides an important ecological defense against grazing by this herbivore.     

An experimental and theoretical analysis of the effect of added weight on the energetics and hydrostatic function of the swimbladder of European sea bass (Dicentrarchus labrax)

We conducted experiments to determine the effect of the increasing ultrasonic/radio transmitter weight on the routine metabolic rate of sea bass. We measured the oxygen consumption (MO₂) of fish tagged externally with a dummy transmitter made of a hollow pipe, the weight of which was adjusted with lead to represent in water 0, 1 and 4% (R_tf) of the animal weight. We then developed a theoretical model to estimate, for a given fish size, the range of added weight that fish can compensate for through swimbladder regulation. When R_tfБ%, MO₂ of untagged and tagged fish did not differ significantly. However, when R_tf reached 4%, fish that carried a tag incurred a significant elevation of oxygen consumption, which represented 28% of their total useable power (or metabolic scope). This result strongly supports the view that a high R_tf ratio contributes to a decrease in available metabolic energy by diverting energy from, e.g., growth or swimming performance. A comparison between the tagged fish and the theoretical model reinforced the hypothesis that, when R_tf attained 4%, the increase in metabolic rate reflected a supplementary and costly swimming effort necessary to maintain vertical position. In this condition, the swimbladder cannot regulate the buoyancy of tagged fish.     

Biomass, photosynthesis and growth of Laminaria saccharina in a high-arctic fjord, NE Greenland

Biomass, photosynthesis and growth of the large, perennial brown alga Laminaria saccharina (L.) Lamour. were examined along a depth gradient in a high-arctic fjord, Young Sound, NE Greenland (74°18'N; 20°14'W), in order to evaluate how well the species is adapted to the extreme climatic conditions. The area is covered by up to 1.6-m-thick ice during 10 months of the year, and bottom water temperature is <0°C all year round. L. saccharina occurred from 2.5 m depth to a lower depth limit of about 20 m receiving 0.7% of surface irradiance. Specimen density and biomass were low, likely, because of heavy ice scouring in shallow water and intensive feeding activity from walruses in deeper areas. The largest specimens were >4 m long and older than 4 years. In contrast to temperate stands of L. saccharina, old leaf blades (2-3 years old) remained attached to the new blades. The old tissues maintained their photosynthetic capacity thereby contributing importantly to algal carbon balance. The photosynthetic characteristics of new tissues reflected a high capacity for adaptation to different light regimes. At low light under ice, or in deep water, the chlorophyll a content and photosynthetic efficiency (!) were high, while light compensation (E_c) and saturation (E_k) points were low. An E_c of 2.0 µmol photons m^-2 s^-1 under ice allowed photosynthesis to almost balance, and sometimes exceed, respiratory costs during the period with thick ice cover but high surface irradiance, from April through July. Rates of respiration were lower than usually found for macroalgae. Annual elongation rates of leaf blades (70-90 cm) were only slightly lower than for temperate L. saccharina, but specific growth rates (0.48-0.58 year^-1) were substantially lower, because the old blades remained attached. L. saccharina comprised between 5% and 10% of total macroalgal biomass in the area, and the annual contribution to primary production was only between 0.1 and 1.6 g C m^-2 year^-1.     

Environmental hypoxia and haemocyanin between-individual variability in Norway lobsters Nephrops norvegicus (L.)

The extent to which exposure to environmental hypoxia (PO₂=8, 6, and 4 kPa, approximately 40%, 30%, and 20% saturation, respectively) resulted in an increase in the concentration of the respiratory pigment haemocyanin ([Hc]) in individuals of the Norway lobster Nephrops norvegicus depended crucially on the initial [Hc] as well as the intensity of the hypoxia itself. While mean [Hc] did increase with decreasing PO₂ (and the variation decreased), for individuals with relatively high initial [Hc] there was no change or even a decrease observed. The greatest hypoxia-related increases in [Hc] were observed in the individuals with the lowest initial [Hc]. Consequently the notion of an 'optimum' [Hc] for a given level of oxygenation was postulated. The changes in [Hc] recorded took place over a considerably shorter time scale (hours rather than days) than has been observed in previous studies. It was not possible to correlate environmental PO₂ and median [Hc] in the field. However, it was interesting that the [Hc] frequency distribution curve for lobsters collected from a site that had been markedly hypoxic (PO₂=8.0 kPa, approximately 40% saturation) until a few days before sampling was almost exactly the same shape (i.e. leptokurtic) and position (i.e. similar median) to that obtained when lobsters were exposed to PO₂=8.0 kPa in the laboratory. Although it would have been desirable to investigate the effect of hypoxia on the frequency distribution of a physiological trait, in this case [Hc], it was not possible due to insufficient sample size, even though the sample sizes employed were considerably larger than those normally used in physiological investigations.     

Induction of larval settlement and metamorphosis of the sea urchin Strongylocentrotus intermedius by glycoglycerolipids from the green alga Ulvella lens

In aquaculture centers of the northern region of Japan, "Nami-ita" (waved polycarbonate plates), on which the green alga Ulvella lens Crouan frat. (Chaetophoraceae: Chaetophorales) was cultured, are used to promote larval settlement and metamorphosis of the sea urchin species Strongylocentrotus intermedius (A. Agassiz) and S. nudus (A. Agassiz). We investigated chemical inducer(s) for larval settlement and metamorphosis of these sea urchins with extracts of U. lens. Bioassay-guided separation of the methanol extract using a combination of column and thin-layer chromatography led to the isolation of several active compounds, the chemical structures of which were determined by spectral and chemical methods. These active compounds were identified as glycoglycerolipids, all comprising several molecular species: sulfoquinovosyl monoacylglycerols (SQMGs), sulfoquinovosyl diacylglycerols (SQDGs), monogalactosyl monoacylglycerols (MGMGs), monogalactosyl diacylglycerols (MGDGs), digalactosyl monoacylglycerols (DGMGs) and digalactosyl diacylglycerols (DGDGs). Among these glycolipids, SQMGs, MGMGs, MGDGs and DGMGs induced larval metamorphosis of the sea urchin S. intermedius. SQMGs and MGDGs induced larval metamorphosis at a concentration of 5 µg ml^-1, whereas SQDGs and DGDGs only induced larval settlement. These glycoglycerolipids are new congeners of chemical inducers to settlement and metamorphosis of planktonic larvae of sea urchins. The findings would provide a better understanding of larval settlement and metamorphosis in sea urchins.     

Natural diets of vertically migrating zooplankton in the Sargasso Sea

The feeding preferences of three common diel vertically migrating zooplankton were investigated from December 1999 to October 2000 at the U.S. JGOFS Bermuda Atlantic Time-Series Study (BATS) station in the Sargasso Sea. Gut content analysis of the copepods Pleuromamma xiphias (Giesbrecht) and Euchirella messinensis (Claus) and of the euphausiid Thysanopoda aequalis (Hansen) indicated that all three species fed on a wide variety of phytoplankton, zooplankton, and detrital material. Diet changes generally reflected seasonal trends in phytoplankton community structure. However, species-specific feeding preferences and differences in feeding selectivity among the three species were evident, and in general agreement with feeding habits predicted from the analysis of mouthpart morphology. The euphausiid T. aequalis fed equally on more different food types compared to both copepod species. The copepod P. xiphias consumed a diverse assemblage of phytoplankton from late winter through the summer (78-93% of gut items, by number, were phytoplankton) and based its diet more strongly on carnivorous feeding in autumn and early winter (31% and 61% of gut items were phytoplankton, respectively). E. messinensis showed the greatest feeding specialization, with a strong preference for pennate diatoms in winter and spring and for coccolithophorids during late summer and fall (constituting 67-93% of gut items by number). All three species consumed diatoms more than other phytoplankton taxa, even though diatoms form only a small fraction of the phytoplankton biomass in the Sargasso Sea. Although the majority of gut items identified were phytoplankton cells, the relative biomass contribution of these small cells may be lower than that of zooplankton and detritus. Zooplankton on which the three species primarily preyed were protozoans and crustaceans, but also included other metazoans such as chaetognaths and cnidarians. Marine snow was also an important component of the diet in all three species, with typically >50% and rarely <20% of the gut content being olive-green debris. Marine snow from larvacean houses was found in the guts of all three species, while E. messinenis appeared to selectively consume marine snow aggregates enriched with bicapitate Nitzschia spp. Large cyanobacteria (>4 µm in diameter) found in guts were also likely consumed with marine snow. The species-specific differences in the diets of these three migrating species suggest that an individual species approach is important in determining how feeding habits affect the structure of pelagic food webs and carbon cycling in the sea. Electronic supplementary material to this paper can be obtained by using the Springer LINK server located at http://dx.doi.org/10.1007/s00227-002-0815-8.     

Studies on pigments,proteins and photosynthetic rates in some mangroves and mangrove associates from Bhitarkanika,Orissa

Pigment contents, proteins and net photosynthesis were investigated in fully developed leaf of 1-year-old seedlings of six mangroves (Bruguiera gymnorrhiza, Rhizophora apiculata) and mangrove associates (Caesalpinia bonduc, Cerbera manghas, Derris heterophylla, Thespesia populnea), collected from Bhitarkanika, located on the east coast of India. Large variations in the photosynthetic rates (P_N) among the six species were observed, ranging from 10.16 µmol CO₂ m^-2 s^-1 in C. bonduc to 15.28 µmol CO₂ m^-2 s^-1 in R. apiculata. The total leaf protein content ranged from 12.09 mg g^-1 dry wt in T. populnea to 51.89 mg g^-1 dry wt in B. gymnorrhiza. The chlorophyll a/b ratio was typically about 3.0 in all the studied species, except C. bonduc (2.8). Photosynthetic rates and chl a/b ratio in the leaves were found to be correlated. Analysis of chlorophyll and xanthophyll spectra suggested: (1) variations in different forms and amounts of carotenes as well as xanthophylls and (2) the presence of high amounts of near-UV-absorbing substances in leaves, particularly in the two mangroves (B. gymnorrhiza, R. apiculata) and a mangrove associate (T. populnea), which appears to be an adaptive feature. Estimation of the chl a/b ratios in isolated thylakoids yielded a low value of 1.8 for R. apiculata and >2.6 for other species. The total protein/chlorophyll ratios in thylakoids varied considerably from 3.14 (D. heterophylla) to 10.88 (T. populnea) among the mangrove associates and from 16.09 to 18.88 between the members of the Rhizophoraceae. The chlorophyll/carotenoid ratios in thylakoids of the six species were more or less similar. The absorption spectra for washed thylakoids of C. manghas and D. heterophylla exhibited absorption characteristics typical for C₃-plant thylakoids. However, thylakoids isolated from R. apiculata, B. gymnorrhiza, C. bonduc and T. populnea exhibited an unusual increase in absorption in the blue region (380-410 nm) of the absorption spectrum. The presence of high-absorbing (in the short-wavelength, near-UV region) pigments appears to be closely associated with the thylakoids in R. apiculata and T. populnea. Our results, therefore, suggest a wide range of variation, not only in protein and pigment contents of photosynthetic tissues, but also in the spectral characteristics and composition of the pigments in mangrove species. An understanding of the nature of these pigments in mangroves and their associates, under their natural conditions and especially in relation to eco-physiological adaptations, is necessary, not only in relation to conservation, but also to allow propagation under different salinity conditions.     

De Novo terpenoid biosynthesis by the dendronotid nudibranch Melibe leonina

Summary. Stable isotope feeding studies using [1,2-¹³C₂]-sodium acetate have demonstrated that 2,6-dimethyl-5-heptenal (1), a putative defensive allomone, is made by the Dendronotid nudibranch Melibe leonina via de novo biosynthesis.     

Consistent association of crenarchaeal Archaea with sponges of the genus Axinella

The association of sponges with microorganisms has been accepted to be a common feature of the phylum Porifera. Herein we describe the association between filamentous Archaea and three Mediterranean species of sponges from the family Axinellidae (Porifera: Demospongiae). Axinella damicornis, A. verrucosa and Axinella sp. harbor a high concentration of filamentous Archaea in the collagen that surrounds the siliceous spicules that form their skeleton. These Archaea have been found in every axinellid specimen studied, regardless of their environment and collection time. The morphology of the filaments has been studied using transmission electron microscopy, and they all show similar characteristics. Their nature has been determined by in situ hybridization experiments and by PCR amplification and sequencing of their 16S DNA. Each sponge species contains a single filamentous archaeal phylotype. The Archaea of the three sponges are closely related to each other and to the marine "group 1" crenarchaeotes. Our findings suggest that this newly described association could be defined as a symbiosis, where biochemical and/or metabolic relationships between the sponge hosts and their symbionts remain to be determined.