The effects of ammonium and phosphate enrichments on clorophyll a,pigment ratio and species composition of phytoplankton of Vineyard Sound

Seawater containing natural phytoplankton populations from Vineyard Sound, USA was enriched in the laboratory with three levels each of ammonium and phosphate and with a combination of ammonium and phosphate which provided three different N:P ratios. The addition of ammonium produced more cells and chlorophyll a than the control or the phosphate enrichments. However, enrichment with ammonium and phosphate, regardless of the N:P ratio, yielded the most cells and chlorophyll a. Thus, nitrogen seems to be the primary limiting nutrient, with phosphate showing secondary limiting effects. The ratios of photosynthetic pigments decreased with the increased chlorophyll a production in the enriched cultures. There were no significant changes in the species composition within the cultures, so that the observed changes in pigment ratio and chlorophyll a content were due to physiological responses.     

Change in the uptake and cellular Si:N ratio in diatoms responding to the ambient Si:N ratio and growth phase

A change in the Si:N ratio of diatom cells during growth was examined for Chaetoceros socialis and Thalassiosira sp., with different initial silicate to nitrate (Si:N) ratios in the media. During exponential growth, C. socialis assimilated silicate and nitrate with a molar ratio of 0.5, independent of the ratio in the media, but after the depletion of nitrate, silicate continued to be taken up, and the Si:N ratio in the stationary phase increased to 2 as a function of the Si:N ratios in the media. In contrast, the ratio of silicate to nitrate taken up by Thalassiosira sp. increased with an increase in the Si:N ratio in the media. The Si:N ratio in the cells during the stationary phase increased in response to an increase in this ratio in the media. The Si:chl  a ratio also increased with the increase in the initial Si:N ratio in the media, while the N:chl  a ratio did not change to a great extent, indicating the changes in the cellular Si:N ratio was derived from changes in the Si content of the cells. These results indicated that the cellular Si:N ratio changed with the Si:N ratio in the medium, and the Si:N uptake ratio during the growth phase was different depending on diatom species. Thus, the dominance of different diatom species may affect nutrient composition and dynamics in the ocean.     

Carbon-nitrogen relations at whole-cell and free-amino-acid levels during batch growth of Isochrysis galbana (Prymnesiophyceae) under conditions of alternating light and dark

Isochrysis galbana Parke, Strain CCAP 927/1, was grown in ammonium-limited batch culture under a 12 h light: 12 h dark illumination cycle. Samples were taken every 12 h over the 26 d period from lag phase through exponential into stationary phase (no net carbon fixation), with more frequent sampling at points of interest. Exponential cell-specific growth rate was 0.3 to 0.4d^-1. Cell division occurred during the dark phase, while cell volume increase, ammonium uptake, and pigment synthesis occurred during the light. Stationary phase cells were small, and the lag phase was long (5 d) even though the C:N ratio had returned from 18 to 6.5 within 2 d, followed by synthesis of chlorophyll a. Net chlorophyll synthesis ceased within 4 d of exhaustion of the nitrogen source. The chlorophyll c: chlorophyll a ratio remained constant during increasing nitrogen deprivation. Biovolume and carotenoids correlated with carbon biomass. Levels of chlorophyll a correlated poorly with carbon fixation and carbon biomass once the nitrogen source had been exhausted. Except after the addition of ammonium to nitrogen-deprived cells (refeeding), the content of intracellular glutamine and the glutamine: glutamate ratio were low during the dark phase, rising to a plateau within the first 1 h of illumination. Refeeding of cells which had only just exhausted the extracellular nitrogen source resulted in a much smaller increase in glutamine than refeeding of nitrogen-starved (stationary-phase) cells. Nitrogen biomass correlated with the presence of an unidentified intracellular amine.     

Saturated uptake kinetics: transient response of the marine diatom Thalassiosira pseudonana to ammonium,nitrate, silicate or phosphate starvation

Changes in the saturated uptake kinetics of the limiting nutrient were followed as Thalassiosira pseudonana (Clone 3 H) batch cultures entered ammonium, nitrate, silicate and phosphate starvation. Cultures starved of ammonium or phosphate developed very high specific uptake capacities over a 24 to 48 h starvation period, due to both decreases in cell quota and increases in uptake rates per cell. In particular, the cell phosphorus quota decreased ca. 8-fold during phosphate starvation and specific uptake rates exceeded 100 d^-1. In contrast, cultures entering nitrate or silicate starvation underwent little or no further cell division, and the uptake capacity declined during starvation. After 24 to 48 h starvation, an induction requirement for uptake of nitrate or silicate was apparent. These responses are consistent with adaptation to the pattern of supply of these nutrients in the field.     

Host feeding and nutrient sufficiency for zooxanthellae in the sea anemone Aiptasia pallida

Nutrient sufficiency of zooxanthellae in the sea anemone Aiptasia pallida cultured in low nutrient seawater depends on the availability of particulate food to the host. Zooxanthellae in anemones unfed for 20 to 30 d exhibited the following characteristics of nutrient deficiency: cell division rates decreased; chlorophyll a content gradually decreased from 2 to <1 pg cell^–1; and C:N ratios increased from 7.5 to 16. Over a 3-mo period, algal populations in unfed anemones gradually decreased, indicating that zooxanthellae were lost faster than they were replaced by division. The mitotic index of zooxanthellae in unfed anemones was stimulated either by feeding the host or by the addition of inorganic N and P to the medium. Whether algae are nutrient-limited in hosts under field conditions has not been examined fully; however, C:N ratios in zooxanthellae from field-collected hosts are slightly higher (9.4 vs 7.5) than in hosts fed to repletion in laboratory cultures. This observation might indicate N limitation in the field.     

Nutrient limitation in the giant clam-zooxanthellae symbiosis: effects of nutrient supplements on growth of the symbiotic partners

The effect of ammonium (5, 10 M N) and phosphate (2, 5, 10 M P) on the growth of the giant clam Tridacna gigas and its symbiotic dinoflagellate Symbiodinium sp. was examined. A 3 mo exposure to these nutritients significantly increased the N or P composition of the soft tissues, as reflected in a corresponding change in C:N:P ratio. Furthermore, exposure to N or N+P markedly increased the amount of soft tissue, but P alone did not, demonstrating that increased availability of inorganic nitrogen enhances tissue growth of the clam host. With addition of N, or N+P, there was a significant increase in the total number of zooxanthellae per clam, with a corresponding decrease in chlorophyll a (chl a) content per zooxanthella. However, only with N+P was there an increase in the zooxanthellae mitotic index. The inverse relationship between zooxanthellae number and chl a per zooxanthella is consistent with phytoplankton studies indicating conditions of nutrient-limitation. Furthermore, the unaffected C:N:P composition of the zooxanthellae and their relatively low specific-growth rates (4 to 10%) also suggest that they are nutrient-limited in vivo. In particular, their high mean C:N:P ratio of 303:52:1 indicates that, relative to C, they are much more depleted in P and less in N than are free-living phytoplankton. Furthermore, polyphosphates (phosphate reserves) were undetectable, and the activity levels of acid phosphatase in the zooxanthellae were relatively high and not influenced by the host's exposure to increased P concentrations in the sea water, implicating the clam host in active regulation of P availability to its symbiotic algae. This is strong evidence that N-limitation of clam zooxanthellae is a function of the availability of ammonium to the symbiosis while, irrespective of nutrient levels in sea water, clam zooxanthellae still show characteristics of P-limitation.     

Effects of silicate depletion on photosynthesis by diatoms in the plume of the Hudson River

Silicate depletion was observed during a bloom of netplankton diatoms. Netplankton chlorophyll a increased over the same salinity range and at the same rate that silicate decreased. Silicate depletion coincided with a decrease in the apparent Si:N uptake ratio as well as a change in the magnitude and diurnal phasing of light saturated photosynthesis (P _m ^B ) by netplankton diatoms. Nanoplankton P _m ^B was unaffected by silicate depletion and increased with temperature. Consequently, nanoplankton P _m ^B eventually exceeded netplankton P _m ^B while netplankton biomass was still increasing relative to nanoplankton biomass.     

Nutrient starvation effects on the allelochemical potency of Alexandrium tamarense (Dinophyceae)

Batch culture experiments were performed to investigate potential effects of nutrient starvation on the allelochemical potency of the toxic dinoflagellate Alexandrium tamarense. Triplicate cultures with reduced nitrate (−N) or phosphate (−P) seed were compared to nutrient-replete (+N+P) cultures. Total depletion of the dissolved inorganic limiting nutrient, reduced cell quotas, changed mass ratios of C/N/P and reduced cell yield clearly indicate that treatment cultures at stationary phase were starved by either N or P, whereas growth cessation of +N+P cultures was probably due to carbon limitation and/or a direct effect of high pH. Pulsed addition of the limiting nutrient allowed −N and −P cultures to resume growth. Lytic activity of A. tamarense as quantified by a Rhodomonas bioassay was generally high (EC₅₀ around 100 cells mL⁻¹) and was only slightly modulated by growth phase and/or nutrient starvation. Lytic activity per cell increased with time in both +N+P and −P cultures but not −N cultures. P-starved stationary-phase cells were slightly more lytic than +N+P cultures, but this difference may be due to increased cell size and/or accumulation of extracellular compounds. In conclusion, only slight changes but no general and major increase in lytic activity in response to nutrient starvation was observed.     

Cell density, chemical composition and toxicity of Chrysochromulina polylepis (Haptophyta) in relation to different N:P supply ratios

The influence of different N:P supply ratios on cell accumulation, chemical composition and toxicity of the marine haptophyte Chrysochromulina polylepis was examined in semi-continuous cultures. A non-axenic strain of C. polylepis was exposed to five different N:P supply ratios (N:P = 1:1, 4:1, 16:1, 80:1 and 160:1, by atoms), in order to create a range of N- and P-limited conditions. The toxicity per cell in C. polylepis was determined on four occasions at steady state cell density using the haemolytic activity of the cells expressed as saponin nanoequivalents. Haemolytic activity was demonstrated in all treatments, and increased in the algae when cell growth was nutrient limited (N:P = 1:1, 4:1, 80:1 and 160:1), compared to cells grown under non-limiting conditions (N:P = 16:1). This occurred regardless of the growth-limiting nutrient (N or P) and became more pronounced as nutrient limitation increased. In P-limited cultures the haemolytic activity per cell increased linearly with the cellular N:P ratio, whereas the N-limited cultures showed an opposite trend. The haemolytic activity per cell showed an inverse relationship with both cellular N and cellular P content. Cells limited by P showed a higher haemolytic activity than cells limited by N. The results suggest that toxicity in C. polylepis is strongly influenced by the physiological state of the algae. This may partially explain the large variability previously observed in the toxicity of C. polylepis blooms. The potential ecological significance of our findings is also discussed. Received: 18 November 1998 / Accepted: 5 July 1999     

Seasonal variation in mineralization rates (C-N-P-Si) of mussel Mytilus edulis biodeposits

To determine seasonal variability in mineralization dynamics of mussel biodeposits, we applied a multiple-element approach measuring mineralization rates of carbon (C), nitrogen (N), phosphorus (P) and silicate (Si) during three periods (March, August and November). The results of this study showed that mineralization rates vary between seasons and between elements and that mineralization dynamics were influenced by both temperature and biodeposit nutrient composition. Mineralization rates were 3.2 ± 0.4 mmol C, 0.17 ± 0.04 mmol N, 0.06 ± 0.02 mmol P and 3.91 ± 3.75 mmol Si per gram biodeposit (DW) per day, which represented 24 % of the particulate organic C and 17 % of the particulate organic N in mussel biodeposits. Seasonal variability was largest for Si mineralization with 60–80-fold higher rates measured in March compared to August and November. This difference is most likely related to the difference in biodeposit nutrient composition. It was furthermore shown that the labile fraction of biodeposits became mineralized after, respectively, 18, 9 and 13 days during the experimental periods in March, August and November. This indicates that temperature enhances biodeposit decomposition with approximately 2–3 times faster turnover at a 10 °C temperature interval (Q ₁₀ ).     

The spring phytoplankton bloom in Lindåspollene,a land-locked Norwegian fjord. II. Biomass and activity of net- and nanoplankton

From February 24 to April 24, weekly samples were collected at fixed depths at one station in Lindåspollene, a land-locked Norwegian fjord. Adenosine triphosphate (ATP), chlorophyll a, phaeophytin, ¹⁴C assimilation, and respiratory activity [electron transport system (ETS) activity] were measured in the net- (>30 m) and nanoplankton. Netplankton contained on the average 48% of the total chlorophyll a and 56% of the ATP, but contributed only 7% to the total carbon assimilation and 11% to the ETS activity. The assimilation numbers for net- and nanoplankton ranged from 0 to 1.2 and from 1.5 to 13.2, respectively. At the oxygen/hydrogen sulphide interface, high concentrations of ATP, but not of chlorophyll a, were found in the nanoplankton fraction. Netplankton algae grew actively only in the first phase of the bloom, and nanoplankton predominated later, apparently due to low nutrient concentrations. During the bloom, Skeletonema costatum made up the main part of the biomass. The number of cells in the chains decreased throughout the bloom, possibly reflecting the lowered silicate content. It appeared that only nanoplankton were grazed by zooplankton, while netplankton sank to the bottom and represented input to the benthos.     

Cell-cell recognition system in gorgonians: description of the basic mechanism

A comparative investigation of the chemical composition of Thalassiosira antarctica var. antarctica vegetative and resting stages revealed C:N and C:chl a ratios to be lower in vegetative cells. These trends primarily reflect vegetative levels of C/cell below, and N/cell and chl a/cell levels above those of spores. There was a change in chemical composition with the initial formation of resting spores, and spores continued to modify their composition while maintained in a cyclic light/dark regime for about one and one half weeks. Most notable was a net increase in carbon and chlorophyll a per cell. Spores then subjected to darkness for over one week appeared to retain most of the carbon and chlorophyll a previously synthesized. These findings support the idea that resting spores enhance the survival capabilities of a species under adverse conditions.     

Effect of bryozoan colonization on inorganic nitrogen acquisition by the kelps Agarum fimbriatum and Macrocystis integrifolia

The effect of bryozoan colonization on inorganic nitrogen acquisition by Agarum fimbriatum Harv. and Macrocystis integrifolia Bory., collected from the west coast of Vancouver Island, British Columbia, Canada, was examined in laboratory experiments during June and July 1992. Pieces of kelp blades that were completely covered on one side by the bryozoans Lichenopora novae-zelandiae Busk or Membranipora membranacea, L., or uncolonized (clean treatment), were used to estimate the rate at which nitrate and ammonium were removed from the surrounding seawater. In addition, the rate of ammonium excretion by bryozoans isolated from their associated kelp was measured and also estimated from the results of the uptake experiments. Values obtained were used to estimate the contribution of ammonium excreted by bryozoans to the total amount of inorganic nitrogen available to the associated kelp. Both bryozoan species reduced the ability of the associated kelp to remove nitrate and ammonium from seawater but provided a source of ammonium to the kelp through excretion. The nitrogen status of colonized and clean kelp disks was determined from the ratio of total particulate carbon to total particulate nitrogen (C:N ratio). The C:N ratios for A. fimbriatum colonized with either L. novae-zelandiae or M. membranacea were similar (C:N=12 to 14), and differences between colonized and clean treatments were not significant. For A. fimbriatum, therefore, the C:N ratio indicates that this species was not nitrogen limited at the time of the present study. In contrast, both colonized and clean disks of M. integrifolia were nitrogen limited, but colonized disks (C:N=19) were significantly less limited by nitrogen than clean disks (C:N=29). Results are discussed in relation to the different environments inhabited by both kelp species and are consistent with the hypothesis that ammonium excreted by bryozoans was an important source of inorganic nitrogen to M. integrifolia, but not to A. fimbriatum, at the time of the study.     

Quantum yield,relative specific absorption and fluorescence in nitrogen-limited Chaetoceros gracilis

Decreases in cell-nitrogen quota resulted in changes in the carbon-based quantum yield of photosynthesis, the chlorophyll a-specific absorption coefficient, and in vivo fluorescence in the marine diatom Chaetoceros gracilis in laboratory experiments performed in 1983 and 1984. The three parameters were independently determined for the two spectral regions dominated by either chlorophyll a or fucoxanthin absorption. As cell-nitrogen quota decreased, the quantum yield for both pigments decreased; the specific absorption coefficient for chlorophyll a and the in vivo chlorophyll a fluorescence excited by each pigment increased. The observed increase in the in vivo fluorescence per chlorophyll a could be partially attributed to the increased specific absorption coefficient for chlorophyll a; the remainder of the fluorescence increase was related to a decline in photosystem activity. Energy transfer efficiency between light-harvesting pigments appeared to be maintained as cell-nitrogen quota decreased. The decrease in a fluorescence index [(F _DCMU-F _O)/F _DCMU] with nitrogen starvation suggested a decrease in Photosystem II activity. These results imply that decreases in reaction center and/or electron-transport system activity were responsible for the decline in rates of photosynthesis under conditions of notrogen deficiency.     

Effects of nitrogen and phosphorus enrichement on in vivo symbiotic zooxanthellae of Pocillopora damicornis

The reef coral Pocillopora damicornis (Linnaeus) was grown for 8 wk in four nutrient treatments: control, consisting of ambient, unfiltered Kaneohe Bay seawater [dissolved inorganic nitrogen (DIN, 1.0 M) and dissolved inorganic phosphate (DIP, 0.3 M)]; nitrogen enrichment (15 M DIN as ammonium); phosphorus enrichment (1.2 M DIP as inorganic phosphate); and 15 M DIN+1.2 M DIP. Analyses of zooxanthellae for C, N, P and chlorophyll a after the 8 wk experiment indicated that DIN enrichment increased the cellular chlorophyll a and excess nitrogen fraction of the algae, but did not affect C cell^-1. DIP enrichment decreased both C and P cell^-1, but the decrease was proportionally less for C cell^-1. the response of cellular P to both DIN and DIP enrichment appeared to be in the same direction and could not be explained as a primary effect of external nutrient enrichment. The observed response of cellular P might be a consequence of in situ CO₂ limitation. DIN enrichment could increase the CO₂ (aq) demand by increasing the net production per unit area. DIP enrichment could slow down calcification, thus decreasing the availability of CO₂ (aq) in the coral tissue.Hawaii Institute of Marine Biology Contribution No. 920     

Silicon and the ecology of marine plankton diatoms. II. Silicate-uptake kinetics in five diatom species

The variation of the rate of silicate uptake with varying silicate concentration in the medium was investigated in short-term experiments with the following marine diatom species:Skeletonema costatum, Thalassiosira pseudonana, T. decipiens, Ditylum brightwellii, andLicmophora sp. The uptake conformed to Michaelis-Menten kinetics only after a correction had been made for reactive silicate that apparently could not be utilized by the diatoms. The magnitude of this correction was in the range of 0.3 to 1.3 g-at Si/l. Mean values of the half-saturation constant of silicate uptake were calculated for the different species. The lowest value was found inS. costatum (0.80 g-at Si/l) and the highest inT. decipiens (3.37 g-at Si/l). Growth limitation by low silicate concentrations could be a cause of species succession in marine plankton-diatom blooms.     

Effect of nutrient enrichment in the field on the biomass, growth and calcification of the giant clam Tridacna maxima

Nutrients were added separately and combined to an initial concentration of 10 μM (ammonium) and/or 2 μM (phosphate) in a series of experiments carried out with the giant clam Tridacna maxima at 12 microatolls in One Tree Island lagoon, Great Barrier Reef, Australia (ENCORE Project). These nutrient concentrations remained for 2 to 3 h before returning to natural levels. The additions were made every low tide (twice per day) over 13 and 12 mo periods for the first and second phase of the experiment, respectively. The nutrients did not change the wet tissue weight of the clams, host C:N ratio, protein content of the mantle, calcification rates or growth rates. However, ammonium (N) enrichment alone significantly increased the total population density of the algal symbiont (Symbiodinium sp.: C = 3.6 · 10⁸ cell clam⁻¹, N = 6.6 · 10⁸ cell clam⁻¹, P = 5.7 · 10⁸ cell clam⁻¹, N + P = 5.7 · 10⁸ cell clam⁻¹; and C = 4.1 · 10⁸ cell clam⁻¹, N = 5.1 · 10⁸ cell clam⁻¹, P = 4.7 · 10⁸ cell clam⁻¹, N + P = 4.5 · 10⁸ cell clam⁻¹, at the end of the first and second phases of the experiment, respectively), although no differences in the mitotic index of these populations were detected. The total chlorophyll a (chl a) content per clam but not chlorophyll a per cell also increased with ammonium addition (C = 7.0 mg chl a clam⁻¹, N = 13.1 mg chl a clam⁻¹, P = 12.9 mg chl a clam⁻¹, N + P = 11.8 mg chl a clam⁻¹; and C = 8.8 mg chl a clam⁻¹, N = 12.8 mg chl a clam⁻¹; P = 11.2 mg chl a clam⁻¹, N + P = 11.3 mg chl a clam⁻¹, at the end of the first and second phases of the experiment, respectively). The response of clams to nutrient enrichment was quantitatively small, but indicated that small changes in inorganic nutrient levels affect the clam–zooxanthellae association. Received: 2 June 1997 / Accepted: 9 June 1997     

Carbon and nitrogen dynamics during growth and degradation of phytoplankton under natural surface irradiance

Under conditions of natural irradiance, the development and decline of a flagellate-dominated phytoplankton population was followed in a coastal North Atlantic pond over a 3 d period in summer 1986. Irradiance negatively affected phytoplankton biomass estimated as chlorophyll a, which decreased during the day at photosynthetically available radiation (PAR) levels above 600 to 1000 mol m^-2s^-1; chlorophyll a increased at PAR values below this threshold. In addition, an inverse relationship was found between changes in chlorophyll a and changes in dissolved inorganic nitrogen, indicating synthesis of nitrogenous biomass mainly at night and degradation mainly during the day, with intense exchanges of material between the particulate and dissolved nitrogen fractions. The natural abundance of ¹³C in particulate matter increased initially, and then remained constant, and was controlled mainly by the ratio -carboxylases activity: ribulose biphosphate carboxylase activity. The hypothesis that the latter enzyme is broken down under high irradiance and is partly responsible for increases in external dissolved nitrogen was rejected.     

Silicon and the ecology of marine plankton diatoms. I. Thalassiosira pseudonana (Cyclotella nana) grown in a chemostat with silicate as limiting nutrient

The small marine plankton diatom Thalassiosira pseudonana Hasle and Heimdal (Guillard's clone 3H) was grown in chemostats with silicate as the limiting nutrient. The calculated maximum growth rates were comparable to those previously reported for this species. The silica content of the diatom shells varied with the growth rate. As the growth rate approached zero, there were still measurable quantities of residual reactive silicate in the medium. In one of the two chemostats used, silicate assimilation by the cells was inefficient due to some unknown internal or external factor. In the other chemostat, statistically calculated half-saturation constants of growth were in the range of 0.5 to 0.8 g—at Si/l, depending on which kind of correction was made for residual silicate. Half-saturation constants of steady-state mean silicate uptake per cell and hour, calculated in a similar fashion, were in the range of 1.4 to 2.6 g—at Si/l. These results indicate that the silicate concentrations causing a reduced silicate uptake by this species in nature do not necessarily result in a correspondingly reduced growth rate. Growth in coastal waters is likely to become seriously limited by a shortage of silicate only when most of the silicate originally present has been removed in the course of a diatom bloom.     

Light absorption by a marine diatom: experimental observations and theoretical calculations of the package effect in a small Thalassiosira species

The relationship between in vivo light absorption efficiency of whole cells and in vitro absorption efficiency of algal pigments has been examined experimentally in the marine diatom Thalassiosira sp. In vitro absorption spectra were obtained for cells disrupted by either ultrasonic treatment or high-pressure shearing stress in a low-temperature (-40°C) pressure cell. A dimensionless measure of the magnitude of the package effect (Q _a ^*), calculated from the ratio of whole-cell to disrupted-cell absorption, ranged from about 0.5 at the blue absorption peak of chlorophyll a (λ=435 nm) to 0.7 at the red chlorophyll a peak (λ=670 nm) to 1.0 at the absorption minimum (λ=600 nm). Cell diameter was found to be an inappropriate measure of size for assessing the magnitude of the package effect. Instead, the effective optical diameter for calculation of intracellular self-shading was found to be less than the cell diameter. This observation is consistent with the fact that most algal pigments are contained within chloroplasts, and that chloroplast volume is necessarily smaller than cell volume.