Lectin-induced enhancement of superoxide anion production by red tide phytoplankton

Chattonella marina, a raphidophycean flagellate, is a highly toxic red tide phytoplankton which causes severe damage to fish farming. Recent studies demonstrated that Chattonella spp. continuously release superoxide anions (O₂ ⁻) while they are living. Heterosigma akashiwo, another raphidophycean flagellate, also produces O₂ ⁻. In the present study, we found that lectins such as concanavalin A (Con A), wheat germ agglutinin (WGA), and castor bean hemagglutinin (CBH) stimulated  C. marina and H. akashiwo to generate enhanced amounts of O₂ ⁻ in a concentration-dependent manner. The lectin-specific sugars potently inhibited the lectin-induced increase of O₂ ⁻ production, suggesting that the effects of lectins are mediated mainly through the interaction of these lectins with carbohydrate moiety present on the flagellate cell surface. In contrast to the potent ability of native Con A (tetravalent), succinylated Con A (divalent) showed only a slight stimulative effect on these flagellates. O₂ ⁻ production was totally inhibited by treatment with proteinase K for 30 min, without affecting the viabilities of flagellates. These results suggest that cell-surface redox enzymes may be involved in O₂ ⁻ production, and such enzymes are responsible for the lectin-stimulation. Received: 21 August 1997 / Accepted: 8 January 1998     

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.     

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.     

Oxygen and carbon stable isotopic profiles of the fan mussel, Pinna nobilis, and reconstruction of sea surface temperatures in the Mediterranean

Stable oxygen and carbon isotope profiles ('¹⁸O_skeletal and '¹³C_skeletal), taken along the direction of growth from the umbo to the shell margin in shells of the pinnid Pinna nobilis, were used to reconstruct sea surface temperatures (SST) in the south-east Mediterranean and ontogenetic records of metabolic CO₂ incorporation. Comparison of the seasonal cycle of SST, predicted from the '¹⁸O_skeletal record of a small (young) rapidly growing pinnid and temperature measured with a continuous in situ recorder showed that P. nobilis calcifies under isotopic equilibrium with surrounding seawater, thus indicating that P. nobilis shells can be used as a reliable predictor of SST. A 10-year SST record for the south-east Mediterranean was reconstructed from the shell profiles of four pinnid shells of different sizes and ages collected in 1995 and 1996. Reliable resolution of the seasonal SST could only be achieved during the first 4 years of shell growth. As the pinnids grew older, the temperature record was poorly resolved because the shell growth had diminished with age, resulting in time-averaging of the record. The amplitude of the generated seasonal temperature cycle compared favourably (DŽ°C) with a long-term temperature record from northern Mediterranean waters. Clear seasonal cycles in '¹³C_skeletal were observed with an amplitude of ~1.0‰, similar to the calculated seasonal changes in '¹³C of seawater (0.6‰) overlying seagrass meadows. An ontogenetic trend towards less positive '¹³C_skeletal values was too large to be attributed to any decrease in '¹³C in seawater resulting from the invasion of anthropogenic CO₂. It is suggested that the temporal changes of '¹³C_skeletal are due to incorporation of respiratory CO₂ into the extrapallial fluid and reflect changes in the metabolic activity of the pinnid rather than changes in the isotopic composition of dissolved inorganic carbon within the surrounding seawater.     

Strategies for carbon acquisition in the red marine macroalga Coccotylus truncatus from the Baltic Sea

Marine macroalgae need carbon-concentrating mechanisms because they have only limited access to CO₂ in their natural environment. Previous studies have shown that one important strategy common to many algae is the activity of periplasmic carbonic anhydrases that catalyse the dehydration of HCO₃^- into CO₂. The latter can then cross the plasma membrane by passive diffusion. We hypothesised that an active (energy-consuming) mechanism might also be involved in the membrane transport of CO₂, as is the case in a number of microalgae. Coccotylus truncatus was chosen as a model organism for this study because it belongs to a group of algae that usually lack direct HCO₃^- uptake: sublittoral red algae. The method used to study carbon uptake was pH drift of the seawater medium surrounding the algae in a closed vessel, with and without the addition of specific inhibitors or proton buffers. Measured parameters included pH, total inorganic carbon and alkalinity of the seawater medium. Our results suggest that, in C. truncatus, periplasmic carbonic anhydrase as well as H⁺ extrusion, probably driven by a vanadate-sensitive P-type H⁺-ATPase (proton pump), are involved in CO₂ uptake. No direct uptake of HCO₃^- was discovered. This paper also presents data on the buffer capacity of several proton buffers and the carbon-uptake inhibitors acetazolamide, 4,4'-diisothiocyano-stilbene-2,2'-disulfonate (DIDS) and orthovanadate in Baltic Sea water with a salinity of 6.5 psu.     

Evaluation of fatty acids as biomarkers for a natural plankton community. A field study of a spring bloom and a post-bloom period off West Greenland

An investigation of the fatty acid composition of a natural arctic plankton community was carried out over two fishing banks located between 63°N and 65°N off the West Greenland coast. Samples for fatty acid analyses, species determination and biomass assessments of the plankton community were taken at the depth of fluorescence maximum. High biomass and diatom dominance during the spring bloom and low biomass and flagellate dominance in the post-bloom period were reflected by the fatty acid profiles. The total amount of fatty acid ranged from 55 to 132 µg l^-1 during the spring bloom and from 1 to 5 µg l^-1 during the post bloom. Analysis of the fatty acids showed that when the plankton was dominated by diatoms of the genera Thalassiosira and Chaetoceros, the proportions of C16:1(n-7) and C20:5(n-3) were correspondingly high. C18s, and particularly C18:1(n-9), were more abundant when the plankton was dominated by small autotrophic flagellates, primarily haptophytes. We found a good positive correlation between the common diatom marker, C16:1(n-7)/C16:0, and the biomass percentage of diatoms (r=0.742, P<0.001), as well as between the biomass percentage of flagellates and total C18 fatty acids (r=0.739, P<0.001). This supports the use of these specific fatty acids and fatty acid ratios as general biomarkers of the plankton community. However, the fatty acids are not specific enough to sufficiently characterise the composition of the plankton community, and microscopical support is needed to verify observed trends.     

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.     

Effects of emersion and elevated haemolymph ammonia on haemocyanin–oxygen affinity of Cancer pagurus

The subtidal crab Cancer pagurus (L.) experiences involuntary periods of emersion associated with practices used in their marketing and distribution. During 24 h emersion, impaired gill function caused an increase of circulating total ammonia (TA=NH₃+NH₄⁺) of 0.35 mmol TA l^-1 (167%). The oxygen-binding characteristics of the haemocyanin of C. pagurus were examined at 10°C in the presence of total ammonia (0.2-1.0 mmol TA l^-1). The haemocyanin-oxygen affinity was decreased in the presence of TA ((logP₅₀/(log[TA]=0.16). Emersion induced significant acidosis and elevated circulating levels of haemolymph TA, lactate and urate, but all had returned to normal levels within 24 h of re-immersion. The accumulation of haemocyanin-modulating substances during 24 h emersion compensated partially (40%) for the effect of the acidosis, but the net effect of the emersion period was a significant decrease in oxygen affinity, corresponding to an increase of P₅₀ (10°C ) from 1.24 kPa (immersed) to 1.96 kPa (24 h emersion). The implications of the findings are considered in terms of the effects and adaptations to emersion.     

Combined effect of temperature and food concentration on the grazing rate of the rotifer Brachionus plicatilis

We developed a predictive relationship to determine the grazing rate of Brachionus plicatilis at given temperatures and food concentrations; this function could be applied to experimental culturing and aquaculture practices. Grazing experiments were conducted at temperatures between 5°C and 40°C and at food concentrations, of the flagellate Isochrysis galbana, ranging between ~0 and 10⁶ ml^-1. In total, 136 grazing rates were determined, using the prey depletion method, for rotifers acclimated to treatments for 0.5 or 4 h. The response of grazing rate to temperature and food concentration was described using a model that combined a rectangular hyperbolic function for food concentration and a sigmoidal function for temperature. Using non-linear curve-fitting methods an equation was obtained: G=(452F)/(159000+F)Ǵ.94/(1+2190002T^-4.35) , where G is the grazing rate (flagellates rotifer^-1 min^-1), F is the food concentration (flagellates ml^-1), and T is temperature (°C). The equation indicates a maximum grazing rate of ~35 prey rotifer^-1 min^-1, above ~4᎒⁵ prey ml^-1 and 25°C.     

Metabolic rates of juvenile scalloped hammerhead sharks (Sphyrna lewini)

Oxygen consumption of juvenile scalloped hammerhead sharks, Sphyrna lewini, was measured in a Brett-type flume (volume=635 l) to quantify metabolic rates over a range of aerobic swimming speeds and water temperatures. Oxygen consumption (log transformed) increased at a linear rate with increases in tailbeat frequency and swimming speed. Estimates of standard metabolic rate ranged between 161 mg O₂ kg^-1 h^-1 at 21°C and 203 mg O₂ kg^-1 h^-1 at 29°C (mean-SD: 189ᆣ mg O₂ kg^-1 h^-1 at 26°C). Total metabolic rates ranged from 275 mg O₂ kg^-1 h^-1 at swimming speeds of 0.5 body lengths per second (L s^-1) to a maximum aerobic metabolic rate of 501 mg O₂ kg^-1 h^-1 at 1.4 L s^-1. Net cost of transport was highest at slower swimming speeds (0.5-0.6 L s^-1) and was lowest between 0.75 and 0.9 L s^-1. Therefore, these sharks are most energy efficient at swimming speeds between 0.75 and 0.9 L s^-1. These data indicate that tailbeat frequency and swimming speed can be used as predictors of metabolic rate of free-swimming juvenile hammerhead sharks.     

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.     

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.     

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.     

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.     

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.     

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.     

The effects of feeding or addition of dissolved inorganic nutrients in maintaining the symbiosis between dinoflagellates and a tropical marine cnidarian

The availability of solid food (Artemia nauplii) and dissolved inorganic nutrients (ammonium, nitrate, phosphate) to the shallow-water marine hydroid Myrionema amboinense was manipulated for 1-8 days in order to investigate their role in the growth of intracellular symbiotic dinoflagellates (zooxanthellae) of the genus Symbiodinium. Symbionts from hydroids collected from the field or maintained under laboratory conditions (25°C, 12 h:12 h light:dark cycle, 80 µE m^-2 s^-1 fluorescent lighting) always exhibited a single peak in mitotic index (MI) at dawn. Symbionts in freshly collected field animals had an MI peak of about 15%. Symbiotic dinoflagellates in hydroids fed Artemia nauplii twice daily in the laboratory maintained this dawn peak of MI between 10% and 15%, but in the absence of feeding or added inorganic nutrients, this peak declined to less than 1% within 2-4 days. In contrast, when hydroids were placed in solutions containing ammonium (20 µM NH₄Cl), nitrate (10 µM NaNO₃), and a combination of ammonium and phosphate (2 µM Na₂HPO₄) immediately after collection, the algal MI remained between 5% and 15% for 4-7 days; the addition of 2 µM phosphate did not increase MI relative to unfed rates. When unfed animals were placed in dissolved nitrogen or fed Artemia, the symbiont MI increased from <1% to 10-17% within 2-3 days; P alone had no effect. However, the increase resulting from added inorganic nutrients was temporary, lasting only 5-7 days. These observations suggest that algal division in the host is maintained indefinitely in the field or by feeding particulate foods twice daily in the laboratory, but the addition of inorganic nutrients alone (ammonium, nitrate and ammonium/phosphate) appeared to support the completion of a maximum of one additional round of cell division. Nutrients required for continued growth and division of symbiotic dinoflagellates are linked to host feeding and host growth; without external food, neither host nor symbiont continue to grow. The same phenomenon is seen in zooxanthellate anemones, clams and corals, where total numbers of symbionts appear to be linked to changes in host-tissue biomass (protein), achieving relatively stable densities in M. amboinense, corals and other cnidarian symbioses, depending on their local environmental conditions. The results of the present study help explain the cellular responses of algal symbionts in reef-dwelling invertebrates to additions of dissolved inorganic nutrients to coral-reef ecosystems.     

Alternate coral–bryozoan competitive superiority during coral bleaching

Bleaching of corals results from the loss of their symbiotic algae (zooxanthellae) and/or pigments. The supply of photoassimilates provided by the zooxanthellae to the coral declines during bleaching and reduces the ability to activate energy-costly processes such as maintenance, growth and reproduction. In the present study we compared the competitive outcomes, expressed as overgrowth and changes in colony sizes of Oculina patagonica (an encrusting Mediterranean stony coral) and the bryozoan Watersipora sp., growing in contact with each other, during and between bleaching events. Year-round observations of tagged colonies showed alternating competitive outcomes: O. patagonica wins over Watersipora sp. between bleaching events, but loses during bleaching events. Using the ¹⁴C-point-labeling technique on coral tissue, we examined intra-colonial translocation of photosynthetic products from the point-tissue labeling towards interaction zones. In non-bleached O. patagonica, competition resulted in preferentially oriented translocation of ¹⁴C products to the interaction zone located up to 8 cm away from the tissue-labeling site. Sites opposite the interaction zone received significantly less labeled photoassimilates compared to the interaction zone. In bleached colonies (40-85% bleached surface area), such translocation did not occur, probably explaining the failure to compete with the encrusting neighbor Watersipora sp. Our findings demonstrate the importance of colonial integration and resource orientation for the competitive superiority of O. patagonica.     

Cross-shelf variation in calanoid copepod production during summer 1996 off the Oregon coast, USA

The fecundity of nine species of adult female calanoid copepods, and molting rates for copepodite stages of Calanus marshallae were measured in 24 h shipboard incubations from samples taken during the upwelling season off the Oregon coast. Hydrographic and chlorophyll measurements were made at approximately 300 stations, and living zooplankton were collected at 36 stations on the continental shelf (<150 m depth) and 37 stations offshore of the shelf (>150 m depth) for experimental work. In our experiments, maximum egg production rates (EPR) were observed only for Calanus pacificus and Pseudocalanus mimus, 65.7 and 3.9 eggs fem^-1 day^-1 respectively, about 95% of the maximum rates known from published laboratory observations. EPR of all other copepod species (e.g., C. marshallae, Acartia longiremis and Eucalanus californicus) ranged from 3% to 65% of maximum published rates. Fecundity was not significantly related to body weight or temperature, but was significantly correlated with chlorophyll a concentration for all species except Paracalanus parvus and A. longiremis. Copepod biomass and production in on-shelf waters was dominated by female P. mimus and C. marshallae, accounting for 93% of the adult biomass (3.1 mg C m^-3) and 81% of the adult production (0.19 mg C m^-3 day^-1). Biomass in the off-shelf environment was dominated by female E. californicus, P. mimus, and C. pacificus, accounting for 95% of the adult biomass (2.2 mg C m^-3) and 95% of the adult production (0.08 mg C m^-3 day^-1). Copepodite (C1-C5) production was estimated to be 2.1 mg C m^-3 day^-1 (on-shelf waters) and 1.2 mg C m^-3 day^-1 (off-shelf water). Total adult + juvenile production averaged 2.3 mg C m^-3 day^-1 (on-shelf waters) and 1.3 mg C m^-3 day^-1 (off-shelf waters). We compared our measured female weight-specific growth rates to those predicted from the empirical models of copepod growth rates of Huntley and Lopez [Am Nat (1992) 140:201-242] and Hirst and Lampitt [Mar Biol (1998) 132:247-257]. Most of our measured values were lower than those predicted from the equation of Huntley and Lopez. We found good agreement with Hirst and Lampitt for growth rates <0.10 day^-1 but found that their empirical equations underestimated growth at rates >0.10 day^-1. The mismatch with Hirst and Lampitt resulted because some of our species were growing at maximum rates whereas their composite empirical equations predict "global" averages that do not represent maximum growth rates.