Phosphorus metabolism of oceanic dinoflagellates: phosphate uptake,chemical composition and growth of Pyrocystis noctiluca

The phosphorus metabolism of Pyrocystis noctiluca Murray (Schuett) 1886 has characteristics which may enhance its potential for success in orthophosphate impoverished waters. The steady-state phosphate uptake rates were equal in the light and dark, and were directly proportional to both the phosphorus cell quota and the cell division rate. In contrast, nutrient-saturated uptake rates were multiphasic, faster in the light than the dark, 2 to 4 orders of magnitude greater than steady-state rates, and were inversely proportional to both the phosphorus cell quota and the cell division rate. These uptake characteristics suggest that P. noctiluca may take up phosphate coincidently at their typically low ambient concentrations as well as to exploit episodic nutrient events in nature. Cell division rates were a hyperbolic function of the ambient orthophosphate concentration. The shortest doubling time was 8.7 d, the phosphate concentration at half the maximum division rate was 0.15 M and the threshold, concentration for cell division was ca 0.05 M PO ₄ ^3- . Division rates of P. noctiluca in the ocean are much faster than predicted from the measured ambient orthophosphate concentrations. Since this dinoflagellate has high naturally occurring alkaline phosphatase activities, and can utilize organic-P compounds, we suggest that organic-P can be as important as orthophosphate in supporting the observed division rates of P. noctiluca in the sea.     

Purification and some properties of β-N-acetyl-D-glucosaminidase from prawn (<Emphasis Type="Italic">Penaeus vannamei</Emphasis>)

The -N-acetyl-D-glucosaminidase (NAGase, EC 3.2.1.52) from prawn (Penaeus vannamei) was purified by extraction with 30% ethanol solution and ammonium sulfate fractionation, then chromatographed on Sephadex G-100 followed by DEAE-cellulose (DE-32) columns. The purified enzyme determined to be homogeneous by polyacrylamide gel electrophoresis (PAGE) and SDS-PAGE. The specific activity of the purified enzyme was 1,560 U mg^–1. Enzyme molecular weight was determined to be 105,000 Da; it contained two subunits of the same mass (45,000 Da). The pI value was calculated to be 4.8 by isoelectric focusing. The optimum pH and optimum temperature of the enzyme for the hydrolysis of pNP--D-GlcNAc (enzyme substrate) were determined to be pH 5.2 and 45°C, respectively. The behavior of the enzyme during hydrolysis of pNP--D-GlcNAc followed Michaelis–Menten kinetics, with K_m=0.254 mM and V_m=9.438 M min^–1, at pH 5.2 and 37°C. The stability of the enzyme was investigated, and the results showed that the enzyme was stable in a pH range from 4.2 to 10.0 and at temperatures <40°C. The effects of metal ions on the enzyme were also studied. Li⁺, Na⁺ and K⁺ had no influence on enzyme activity. Mg²⁺, Ca²⁺ and Mn²⁺ activated the enzyme, while Ba²⁺, Zn²⁺, Co²⁺, Cd²⁺, Hg²⁺, Pb²⁺ Cu²⁺, Fe³⁺ and Al³⁺ showed various degrees of inhibitory effects on the enzyme.Communicated by O. Kinne, Oldendorf/Luhe     

Calcification in the maerl coralline alga Phymatolithon calcareum: Effects of salinity and temperature

The coralline alga Phymatolithon calcareum was dredged from 13 m in the Kattegatt, Baltic Sea, in December, 1980, and its rate of calcification was measured by ⁴⁵Ca⁺⁺-uptake methods. Light-saturated calcification rates at 5°C ranged from 15.8 g CaCO₃ g^-1 dry wt h^-1 for the basal parts of the plants to 38.7 g CaCO₃ g^-1 dry wt h^-1 for the tips. These age gradients were not apparent when calcification rates were expressed on the basis of surface area. Experiments with salinity (10, 20, 30) and temperature (0°, 5°, 10°, 20°C) indicated that optimum conditions for calcification were at 30 S and at temperatures above 10°C. Salinity had a greater influence on calcification rate than did temperature, and there was a positive relationship between salinity and calcification rate at all temperatures. In 6 mo old cultures, salinity was again the important factor, with all plants remaining healthy at 30 except those at the highest temperature (20°C). These trends, and the low calcification rates at 10S (4.6 g CaCO₃ g^-1 dry wt h^-1 at 5°C to 8.6 g CaCO₃g^-1 dry wt h^-1 at 20°C) suggest that low salinity may be the explanation for the general absence of P. calcareum from the brackish waters of the Baltic Sea. Short-term experiments in which salinity was kept constant while Ca⁺⁺ concentration was altered, and experiments in which salinity was varied and Ca⁺⁺ concentration kept constant, suggest that it is the calcium ion concentration and not salinity per se which affects calcification rates.     

Respiration and excretion by the ctenophore Mnepiopsis leidyi

Respiration (dissolved oxygen and carbon dioxide) and excretion (dissolved organic carbon, inorganic and organic nitrogen and phosphorus) rates were measured for a variety of sizes of Mnemiopsis leidyi over a temperature range of 10.3° to 24.5°C. Both respiration and excretion rates were a direct linear function of animal weight and very temperature sensitive (Q₁₀4). Oxygen uptake ranged from 155 to 489 g at O/(g dry weight) day^-1 and carbon dioxide release from 43 to 166 M. Organic carbon made up about 38% of the total carbon released. Inorganic nitrogen excretion, exclusively in the form of ammonium, comprised 54% of the total nitrogen release and ranged from 10 to 36 M NH₄/(g dry weight) day^-1. Average release of dissolved primary amines (expressed as glycine equivalents) equaled 43% of the organic nitrogen fraction. Inorganic phosphorus release ranged from 2.0 to 4.9 M/(g dry weight) day^-1 and made up about 72% of the total phosphorus loss. The turnover of elements in the body was calculated as 5 to 19% per day for carbon and nitrogen, depending on the temperature, and an even higher 20 to 48% per day for phosphorus. These values are comparable to rates observed for small, active zooplankton.     

Wet deposition chemistry studies at Suva, Fiji, a remote tropical island site in the south Pacific

A field project encompassing wet-only rainwater sampling was initiated as a bilateral Fiji/Australia activity. Normally, biweekly samples were collected, using a wet-only rainwater sampler, and analysed for H⁺, Na⁺, K⁺, Mg²⁺, NH₄ ⁺, Cl^–, NO₃ ^–, SO₄ ^2–, PO₄ ^3-, methane sulphonic acid, oxalic acid, formic acid and acetic acid. The pH of the rainwater ranged between 5.730 and 4.480 with an average value of 5.176, slightly lower than the pH of unpolluted rainwater saturated with atmospheric CO₂(pH = 5.650). Na⁺and Cl^–were the major ions with average concentrations of 98.15 M and 109.57 M respectively. There is an excellent correlation between the cation sum (average 147.71 eq L^-1) and the anion sum (average 142.12 eq L^-1) attesting to the quality of the data generated. This paper presents the detailed results of the study for a relatively clean remote island site in Suva, Fiji, latitude 18° 09 S, longitude 178° 27 E, height 6 m, and outlines prospects for further work.     

Photosynthetic carbon acquisition in the red alga Gracilaria conferta

In this continuing study on photosynthesis of the marine red alga Gracilaria conferta, it was found that ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) in crude extracts had a K _m (CO₂) of 85 M. Since seawater contains only ca. 10 M CO₂, it appears that this alga must possess a CO₂ concetrating system in order to supply sufficient CO₂ to the vicinity of the enzyme. Because this species is a C₃ plant (and thus lacks the C₄ system for concentrating CO₂), but can utilize HCO₃ ^- as an exogenous carbon source, we examined whether HCO₃ ^- uptake could be the initial step of such a CO₂ concetrating system. The surface pH of G. conferta thalli was 9.4 during photosynthesis. At this pH, estimated maximal uncatalyzed HCO₃ ^- dehydration (CO₂ formation) within the unstirred layer was too slow to account for measured phostosynthetic rates, even in the presence of an external carbonic anhydrase inhibitor. This observation, and the marked pH increase in the unstirred layer following the onset of light, suggests that a HCO₃ ^- transport system (probably coupled to transmembrane H⁺/OH^- fluxes) operates at the plasmalemma level. The involvement of surface-bound carbonic anhydrase in such a system remains, however, obscure. The apparent need of marine macroalgae such as G. conferta for CO₂ concentrating mechanisms is discussed with regard to their low affinity of Rubisco to CO₂ and the low rate of CO₂ supply in water. The close similarity between rates of Rubisco carboxylation and measured photosynthesis further suggests that the carboxylase activity, rather than inorganic carbon transport and intercoversion events, could be an internal limiting factor for photosynthetic rates of G. conferta.     

Effects of tidal fluctuations of salinity on pericardial fluid composition of the American oyster Crassostrea virginica

Crassostrea virginica Gmelin were subjected to simulated tidal fluctuations of salinity, and the subsequent effects on osmotic and ionic composition of the pericardial fluid, body water and valve movements were investigated. Ambient salinity fluctuation patterns of 20-10-20, 15-10-15 and 10-5-10 were simulated during 24.8-h periods. An additional 10-5-10 S experiment was performed using a dilution water approximating the ionic composition of Mississippi River water with regard to Mg⁺⁺, Ca⁺⁺ and SO ₄ ⁼ , instead of deionized water. Finally the effects of a 2-week diurnal fluctuation pattern between 20 and 10 S were investigated with respect to pericardial fluid composition. Pericardial fluid osmolality, concentrations of Cl^-, Na⁺, Mg⁺⁺, K⁺, Ca⁺⁺ and ninhydrin-positive substances (NPS) were analyzed periodically throughout all experiments. Pericardial fluid osmolality was slightly hyperosmotic as ambient water salinity decreased during a cycle, and then became slightly hyposmotic as ambient salinity increased. In the 2-week experiment, pericardial fluid osmolality tracked ambient seawater closely through Day 5, but became more intermediate between high and low seawater values as the experiment progressed. Similar patterns during fluctuations of salinity were observed for Na⁺, Cl^-, Mg⁺⁺ and Ca⁺⁺. Pericardial fluid K⁺ levels did not track ambient seawater as closely as did other ions. The ionic composition of dilution water had little effect on the osmotic or ionic response of the oyster's pericardial fluid. Pericardial fluid NPS level varied inversely with salinity during the 20-10-20 cycle. During the longterm fluctuation experiment, NPS values gradually decreased over the 2-week period compared to constant salinity control values. Percent body water also varied inversely with ambient salinity. Solute movement accounted for most of the change in pericardial fluid osmolality during the simulated cycles with water movement responsible for 1 to 11%. Water movement contributed more to the change of pericardial fluid osmolality during the decreasing salinity phase than the increasing phase of a given cycle. During 20-10-20 S cycles, oyster valves remained open 56% of the time (n=23). In contrast, when salinity was abruptly changed from 20 to 10 within 5 min, valve closure occurred in 4.8±0.3 min (n=20). Valves did not reopen for 19.3±1.2 h (n=15).     

The common jellyfish Aurelia aurita: standing stock,excretion and nutrient regeneration in the Kiel Bight,Western Baltic

The population dynamics, ammonia and inorganic phosphate excretion, and nutrient regeneration of the common jellyfish Aurelia aurita was investigated from 1982 to 1984 in the Kiel Bight, western Baltic Sea. During summer 1982, medusae abundance ranged between 14 and 23 individuals 100 m^-3, biomass was estimated at about 5 g C 100 m^-3 and the mean final diameter of individuals was 22 cm. Abundance, based on numbers, in 1983 and 1984 was an order of magnitude lower; biomass was less than 2 g C 100 m^-3 and jellyfish grew to 30 cm. During the summers of 1983 and 1984, A. aurita biomass constituted roughly 40% of that of the total zooplankton>200 m. In 1982, for which zooplankton data were lacking, it was assumed that medusae biomass was greater than that of all other zooplankton groups. Total ammonia excretion ranged between 6.5 and 36 mol h^-1 individual^-1, whereas inorganic phosphate release was 1.4 to 5.7 mol h^-1 individual^-1. Allometric equations were calculated and exponents of 0.93 for NH₄–N release and 0.87 for PO₄–P excretion were determined. Nitrogen and phosphorus turnover rates were 5.4 and 14.6% d^-1, respectively. In 1982, the medusae population released 1 100 mol NH₄–N m^-2 d^-1, about 11% of the nitrogen requirements of the phytoplankton. The inorganic phosphate excretion (150 mol m^-2 d^-1) sustained 23% of the nutrient demands of the primary producers. In the other two years the nutrient cycling of the medusae was much less important, and satisfied only 3 to 6% of the nutrient demands. It is suggested that in some years A. aurita is the second most important source of regenerated nutrients in Kiel Bight, next to sediment.     

Stable isotope and biochemical fractionation in the marine pelagic food chain: the jellyfish Pelagia noctiluca and net zooplankton

In our field study we analyzed the C and H isotopic and biochemical (C, N, P, protein, lipid, carbohydrate) composition of the jellyfish Pelagia noctiluca (collected from the Gulf of Trieste in 1985 to 1986) and its presumed diet-net zooplankton. The mean ¹³C (-18.8) and D (-58.4) ratios of P. noctiluca showed enrichment in heavy isotopes relative to net zooplankton (2 for carbon and 30 for hydrogen). Both the jellyfish and net zooplankton were characterized by a linear correlation between ¹³C and D. C. N, P, protein, lipid, and carbohydrate contents of P. noctiluca were low on a dry weight basis as compared to net zooplankton. Significantly lower C:N and C:P ratios were found in jellyfish indicating a greater loss of carbon relative to nitrogen and phosphorus along the passage to a higher trophic level. Isotopic and biochemical evidence indicate that, though collected in nearshore waters, P. noctiluca depended on autochthonous marine organic matter.     

Photosynthetic carbon acquisition in the red algaGracilaria conferta

Photosynthetic properties of the common red algaGracilaria conferta, collected from the eastern Mediterranean Sea were investigated in 1989, in order to begin evaluating its adaptative strategies with regard to the inorganic carbon composition of seawater, and to test whether the alleged C₄ photosynthesis of anotherGracilaria species is common within the genus. Net photosynthetic rates ofG. conferta were, under ambient conditions of inorganic carbon (ca. 10µM, CO₂ and 2.2 mM HCO ₃ ^- ), not sensitive to O₂ over the range 10 to 300µM, and the CO₂ compensation point was low (ca. 0.005µM). Ribulose-1,5-bisphosphate carboxylase/oxygenase was the major carboxylating enzyme, with a crude extract activity of 175µmol CO₂ g^–1 fresh wt h^–1 while phosphoenolpyruvate carboxylase and phosphoenolpyruvate carboxykinase were present at 70 and 20%, respectively, of that activity. No activities of the decarboxylases NAD-and NADP-malic enzyme could be detected. The¹⁴C pulse-chase incorporation pattern showed thatG. conferta fixes inorganic carbon via the photosynthetic carbon reduction cycle only, with no evidence for photosynthetic C₄ acid metabolism. Photosynthesis at the natural seawater pH of 8.2 was, at 25°C and saturating light, saturated at the ambient inorganic carbon concentration of 2.5 mM. It is proposed that, under ambient inorganic carbon conditions, a CO₂ concentrating system other than C₄ metabolism provides an internal CO₂ concentration sufficient to suppress the O₂ effect on ribulose-1,5-bisphosphate carboxylase/oxygenase and, thus, on photorespiration, in a medium where the external free CO₂ concentration is lower than theK _m(CO₂) of the carboxylating enzyme. Since inorganic carbon, under natural saturating light conditions, seems not to be a limiting factor for photosynthesis ofG. conferta, it likely follows that other nutrients limit the growth of this alga in nature.     

Phosphatase activity in Eutrophic Tokyo Bay

Alkaline phosphatase activity in seawater samples taken from Tokyo Bay was measured by both spectrophotometric and fluorometric methods. A stratified distribution pattern of the enzyme was observed in August, a vertically mixed pattern in December. The distribution of phosphatase activity in the eutrophic seawater was paralleled by variations in other parameters, such as viable counts of bacteria, chlorophyll a content, inorganic and total phosphorus concentrations, amounts of seston, particulate deoxyribonucleic acids (DNA) and protein. A significant correlation between phosphatase activity and these measurements indicated that the enzyme was a good indicator of the degree of eutrophication. The positive relationship between phosphatase and inorganic phosphorus indicates that enzyme activity was not inhibited at inorganic phosphorus levels present in the bay and that production of phosphatase by microorganisms inhabiting the bay was not repressed at the inorganic phosphorus levels in the bay. Culture experiments revealed that the formation of repressible phosphatase by bacteria isolated from the bay was not affected by the inorganic phosphorus levels in the bay.     

Decomposition of organic and solubilisation of inorganic phosphorus compounds by bacteria isolated from a marine sandy beach

The ability to decompose organic and to solubilise inorganic phosphorus compounds was studied in heterotrophic bacteria isolated from a sandy beach in Sopot, southern Baltic coast. Bacteria able to hydrolyse DNA and phytin were most numerous, while only a small percentage of the studied strains was able to depolymerise glycerophosphate. The ranking of the potential of the studied bacteria to solubilise metallic phosphate salts as follows: calcium hydrogen phosphate>ferric phosphate>magnesium phosphate>calcium phosphate>aluminium phosphate. There were marked differences in the level of depolymerisation of organic compounds of phosphorus and solubilisation of inorganic phosphorus between bacteria inhabiting different parts of the studied beach, while there were generally no differences between the surface and subsurface sand layers. The activity of alkaline phosphatase was higher than that of acid phosphatase.Communicated by O. Kinne, Oldendorf/Luhe     

Seasonal growth in Laminaria longicruris: Relations with dissolved inorganic nutrients and internal reserves of nitrogen

Observations have been made on seasonal fluctuations in dissolved inorganic nutrients, internal reserves of nitrogen and growth rates in Laminaria longicruris. The onset of winter growth in shallow-water stations (6 and 9 m) correlated well with improved dissolved nitrate conditions in the sea. During the winter, reserves of NO ₃ ^- were accumulated by the plants and reached maximum values of 150 moles per g fresh weight in March. This represents a concentration factor of approximately 28,000 over the ambient levels, or an internal nitrogen reserve of 2.1% of the dry weight of the tissue. Depletion of this nitrogen pool followed the disappearance of the external NO ₃ ^- with a lag period of up to 2 months. Rapid kelp growth was measured during this period. Reserves of organic nitrogen also reached maximum values in March and declined slowly throughout the summer into autumn. It is suggested that the combined inorganic and organic nitrogen reserves sustain the rapid growth rates into July and at reduced rate through the late summer. Fertilization of an experimental perimental kelp bed with NaNO₃ increased the internal plant reserves of NO ₃ ^- and produced a much improved summer growth rate. The enriched plants developed very small reserves of carbohydrate during the rapid summer growth phase.NRCC No. 15549.     

Some properties of calcium-activated adenosine triphosphatase from the hermatypic coralGalaxea fascicularis

Samples of the hermatypic coralGalaxea fascicularis were collected between April 1987 and April 1990 from coral reefs off Singapore (103 °45E; 1 °13N). Ca²⁺-activated adenosine triphosphatase (ATPase) activity was detected in the plasma-membrane-enriched heavy microsomal fraction ofG. fascicularis. The high affinity component hadKm andVmax values of 0.0021 mM and 0.050 µmol Pi mg^–1 protein min^–1, respectively; corresponding values for the low affinity component were 0.15 mM and 0.85 µmol mg^–1 protein min^–1. The activity of the high affinity component was inhibited 80 and 50%, respectively, by the anticalmodulin drugs calmidazolium and chlorpromazine. The low affinity component of the Ca²⁺-ATPase may represent activities of alkaline phosphatase, Ca²⁺-ATPase from membranes of mitochondria and endoplasmic reticulum, or calmodulin-dissociated plasma membrane Ca²⁺-ATPase resulting from the removal of Ca²⁺ by EDTA during the isolation process. The high affinity Ca²⁺-ATPase is probably the enzyme responsible for Ca²⁺ extrusion from the cells ofG. fascicularis. The high and low affinity components of this Ca²⁺-ATPase could use ATP and ADP as substrates. Maximum activities of both components were registered at pH 7 and at 45°C. Ruthenium red, a specific inhibitor of Ca²⁺-ATPase, inhibited the activities of the high and low affinity Ca²⁺-ATPase by 100 and 60%, respectively. Inhibition of the activities of both components was also observed with sulphydryl reagents (PCMB and mersalyl). However, DCMU, diamox, dinitrophenol, iodoacetate, fluoride, cyanide, ouabain, oligomycin B and L-phenylalanine had no effect on the enzyme activities.     

Water and salt balance of halibut eggs and larvae (Hippoglossus hippoglossus)

Eggs of halibut [Hippoglossus hippoglossus (L.)] have a negative buoyancy in sea water of 35 S, in contrast to eggs of most flatfish species. The cause of this was investigated. The osmolality of the yolk is 350–420 mOsm during embryonic development. This is within the range for marine teleost serum and for yolk of pelagic eggs. Concentrations of major inorganic ions are comparable with those of pelagic plaice eggs [Pleuronectes platessa (L.)]. The values for Na⁺, K⁺ and Cl^- are 6, 85, and 64 mmol · (1 H₂O)^-1 after fertilization, and at the time of hatching the corresponding values are 17, 11, and 80. Large amounts of other inorganic constitutents are excluded for osmotic reasons. Malfunction in the regulation of osmolality or of inorganic constituents is thus unlikely to be the cause of negative buoyancy. The relative dry weight of the chorion (egg shell) in halibut eggs is less than in several pelagic egg types, excluding the chorion as the main contributor of negative buoyancy. It is concluded that a high content of organic matter in the rest of the egg is the cause of the negative buoyancy.     

锌在不同磷源条件下对铜绿微囊藻生长与产毒的影响

针对磷和微量元素对藻类生长的共同作用,研究不同磷源条件下锌对藻细胞生长与产毒的影响。实验选用铜绿微囊藻为藻种,分别以无机磷磷酸氢二钾(K_2HPO_4)、小分子有机磷甘油磷酸钠(NaGly)和大分子有机磷卵磷脂(LEC)为磷源,研究不同锌(Zn~(2+))含量对藻细胞的藻密度、碱性磷酸酶活性(alkaline phosphatase activity,APA)以及胞内藻毒素(MC-LR)的影响。研究发现:以NaGly为磷源时微量元素锌对藻细胞生长的促进效果显著,而以K_2HPO_4或LEC为磷源时,锌含量的变化对藻细胞生长无显著影响。APA不仅与磷源有关而且与锌含量相关,以LEC为磷源时的APA显著高于以K_2HPO_4或NaGly为磷源时的APA,且锌含量越低APA越低,以K_2HPO_4为磷源时锌含量越低APA越高,而锌对以NaGly为磷源时的APA几乎没有影响。磷源与微量元素锌对藻细胞的产毒均产生影响,NaGly有利于藻毒素的产生; LEC不利于藻细胞的产毒,但锌含量越低藻细胞的产毒量越多。综上所述,磷源与微量元素锌共同作用对藻细胞的生长与产毒产生影响,小分子有机磷NaGly与锌的效果显著。     

Effect of phosphate and ammonium levels on photosynthetic and respiratory responses of the red alga Gracilaria verrucosa

Gracilaria verrucosa (Hudson) Papenfuss exposed to nutrient enriched media (0.1 mM PO₄; 1.0 mM NH ₄ ⁺ ) by pulse feeding 2 h every third day for a period of 5 wk at 20°C and 25–30 salinity showed significantly higher rates of photosynthesis regardless of photon flux density correlated with increased pigment levels. Algae in nonenriched media showed significantly higher levels of soluble carbohydrates and decreased levels of phycoerythrin and chlorophyll a. Photosynthetic and respiratory responses to temperature 15°, 25°, 30°C and salinity (15, 25, 30 S) combinations indicate broad tolerances by both nutrient enriched and non-nutrient enriched algae. Photosynthetic and respiratory rates were highest at the high temperatures. Pulse-fed algae had significantly higher photosynthetic rates than non-nutrient enriched plants at all temperature and salinity combinations. Non-nutrient enriched algae had significantly higher respiratory rates than nutrient enriched algae at only 30°C and 15. The respiratory rates of both nutrient enriched and non-nutrient algae decreased under combinations of higher temperatures and salinities. G. verrucosa, grown without nutrients, has lower tolerances to environmental stresses.     

Regulation of growth in Laminaria digitata: use of in-vivo nitrate reductase activities as an indicator of nitrogen limitation in field populations of Laminaria spp.

The seasonal variation in growth rate of a population of Laminaria digitata (Huds.) Lamour growing at Arbroath, Scotland was studied between August 1981 and September 1982, and was found to follow the biphasic annual cycle typical of this genus. Growth rates were maximum (0.3 cm cm^-1 mo^-1) in early June and minimum (0.05 cm cm^-1 mo^-1) between September and January. An analysis of the relationship between the seasonal changes in environmental factors (inorganic nitrogen concentrations, irradiance and temperature) with those of growth rate and the accumulation or mobilisation of cellular reserves of carbohydrates and nitrate, indicated that growth was nitrogen-limited between June and October and light-limited (with a possible co-involvement of temperature) for the remainder of the year. These conclusions were supported by the seasonal changes in the ratio of actual: potential in-vivo nitrate reductase activities in L. digitata, thus confirming the suitability of this technique for monitoring the occurrence of nitrogen limitation in Laminaria spp. The seasonal changes in blade nitrate reductase activities closely followed those of growth rate, with maximum activities [0.3 mol NO ₃ ^- reduced g^-1 (wet wt) h^-1] being present in late May and minimum levels [0.01 mol NO ₃ ^- reduced g^-1 (wet wt) h^-1] occurring between November and March. The correlation observed between nitrate reductase activities and growth rate is consistent with the ability of Laminaria spp. to store excess inorganic nitrogen, available during winter and early spring, as NO ₃ ^- , and with the requirement to conserve enzyme protein during the summer period of nitrogen limitation.     

Acid and alkaline phosphatase activities in the clam Ruditapes philippinarum

Acid and alkaline phosphatase activities have been partially characterized in Ruditapes philippinarum (Adams and Reeve, 1850). Two activity peaks at pH=4.5 and pH 10.5 were detected in the gill, digestive gland, mantle, siphon and foot. Acid phosphatase activity was higher than that of alkaline phosphatase. The highest activity for both enzymes was observed in the digestive gland and, in decreasing order, the gill, foot, siphon and mantle. Alkaline phosphatase activity was similar in the mantle, siphon and foot. K _m values were determined for both enzymes in the gill and digestive gland. Hill coefficients were near 1, indicating no allosteric behaviour for either enzyme in the two organs. The optimum temperature was the same for acid phosphatase in both gill and digestive gland (50 °C), while for alkaline phosphatase it differed for these two organs (gill, 40 °C; digestive gland, 35 °C). The apparent activation energy was obtained from Arrhenius plots, and ranged from 8.61 kcal/mol for alkaline phosphatase in the gill, to 10.84 kcal/mol for acid phosphatase in the digestive gland. The effects of metals (1 mM conc) on both enzyme activities were assayed in vitro. Hg strongly inhibited the enzyme activities in the gill and digestive gland, probably because of its affinity for the sulphydryl group. Histochemically, acid phosphatase in the gill was located in a granular form throughout the gill cells, but was undetectable in the ciliate epithelium of the gill filaments. Alkaline phosphatase was located in the gill skeleton. Clam size and phosphatase activities were inversely related, probably reflecting a decrease in shell deposition with inereasing size. As a function of season, both enzymes were present in lowest amounts in winter, when undifferentiated sex cells were predominant in the germinative epithelium, and highest in summer, when ripe individuals of both sexes were more frequent.     

Adenylate cyclase of Mytilus galloprovincialis posterior adductor muscle: basal properties and sensitivity to serotonin

The presence and characteristics of a membrane-bound adenylate cyclase from Mytilus galloprovincialis Lmk posterior adductor muscle have been investigated. The enzyme has a Michaelis constant (K _m) of 0.38 mM at 20°C and requires divalent cations (Mg²⁺/Mn²⁺) for its activity. Optimal GTP concentrations are between 10^-5 and 10^-4 M. The non-hydrolizable GTP-analogues GMPpNHp and GTPS increase the activity of the enzyme four- to ten-fold. Sodium fluoride stimulates the enzymatic activity seven- to eight-fold. Forskolin increases the enzymatic activity two- to three-fold. Serotonin stimulates the adenylate cyclase activity in a dose-dependent manner. These experiments were performed with mussels collected from the estuary of Muros, Spain, in 1990.