The influence of light on copper‐limited growth of an oceanic diatom,Thalassiosira oceanica (Coscinodiscophyceae)

Thalassiosira oceanica (CCMP 1005) was grown over a range of copper concentrations at saturating and subsaturating irradiance to test the hypothesis that Cu and light were interacting essential resources. Growth was a hyperbolic function of irradiance in Cu‐replete medium (263 fmol Cu′ · L^?1) with maximum rates achieved at 200 μmol photons · m^?2 · s^?1. Lowering the Cu concentration at this irradiance to 30.8 fmol Cu′ · L^?1 decreased cellular Cu quota by 7‐fold and reduced growth rate by 50%. Copper‐deficient cells had significantly slower (P < 0.0001) rates of maximum, relative photosynthetic electron transport (rETR_max) than Cu‐sufficient cells, consistent with the role of Cu in photosynthesis in this diatom. In low‐Cu medium (30.8 fmol Cu′ · L^?1), growth rate was best described as a positive, linear function of irradiance and reached the maximum value measured in Cu‐replete cells when irradiance increased to 400 μmol photons · m^?2 · s^?1. Thus, at high light, low‐Cu concentration was no longer limiting to growth: Cu concentration and light interacted strongly to affect growth rate of T. oceanica (P < 0.0001). Relative ETR_max and Cu quota of cells grown at low Cu also increased at 400 μmol photons · m^?2 · s^?1 to levels measured in Cu‐replete cells. Steady‐state uptake rates of Cu‐deficient and sufficient cells were light‐dependent, suggesting that faster growth of T. oceanica under high light and low Cu was a result of light‐stimulated Cu uptake.     

EXTREMELY HIGH GROWTH RATE OF THE SMALL DIATOM CHAETOCEROS SALSUGINEUM ISOLATED FROM AN ESTUARY IN THE EASTERN SETO INLAND SEA,JAPAN1

Small single‐celled Chaetoceros sp. are often widely distributed, but frequently overlooked. An estuarine diatom with an extremely high growth potential under optimal conditions was isolated from the Shinkawa‐Kasugagawa estuary in the eastern part of the Seto Inland Sea, western Japan. It was identified as Chaetoceros salsugineum based on morphological observations. This strain had a specific growth rate of 0.54 h^?1 at 30°C under 700 μmol · m^?2 · s^?1 (about 30% of natural maximal summer light) with a 14:10 L:D cycle; there was little growth in the dark. However, under continuous light it grew at only 0.35 h^?1 or a daily specific growth rate of 8.4 d^?1. In addition, cell density, chlorophyll a, and particulate organic carbon concentrations increased by about 1000 times in 24 h at 30°C under 700 μmol · m^?2 · s^?1 with a 14:10 L:D cycle, showing a growth rate of close to 7 d^?1. This very rapid growth rate may be the result of adaptation to this estuarine environment with high light and temperature. Thus, C. salsugineum can be an important primary producer in this estuary in summer and also an important organism for further physiological and genetic research.     

PHYSIOLOGICAL RESPONSES OF ANABAENA VARIABILIS (CYANOPHYCEAE) TO INSTANTANEOUS EXPOSURE TO VARIOUS COMBINATIONS OF LIGHT INTENSITY AND TEMPERATURE1

Light intensity and temperature interactions have a complex effect on the physiological process rates of the filamentous bluegreen alga Anabaena variabilis Kütz. The optimum temperature for photosynthesis increased with increasing light intensity from 10°C at 42 μE·m^?2·s^?1 to 35°C at 562 μE·m^?2·s^?1. The light saturation parameter, I_K, increased with increasing temperatures. The maximum photosynthetic rate (2.0 g C·g dry wt.^?1·d^?1) occurred at 35°C and 564 μE·m^?2·s^?1. At 15°C, the maximum rate was 1.25 g C·g dry wt.^?1·d^?1 at 332 μE·m^?2·s^?1. The dark respiration rate increased exponentially with temperature. Under favorable conditions of light intensity and temperature the percent of extracellular release of dissolved organic carbon was less than 5% of the total C fixed. This release increased to nearly 40% under combinations of low light intensity and high temperature. A mathematical model was developed to simulate the interaction of light intensity and temperature on photosynthetic rate. The interactive effects were represented by making the light-saturation parameters a function of temperature.     

Process optimization and modeling for the cultivation of Nannochloropsis sp. and Tetraselmis striata via response surface methodology

The aim of this study was to determine the optimal physical process conditions for the cultivation of locally isolated strains of Nannochloropsis sp. and Tetraselmis striata to achieve maximum growth rate. It was essential to evaluate biomass production at different agitation rates, light intensities, and temperature levels. Central composite design and response surface methodology were applied to design the experiments and optimize the cultivation process for Nannochloropsis sp. and T. striata. The specific growth rate of 0.250 d^?1 was obtained for Nannochloropsis sp. cells under the light intensity of 54 μmol photons · m^?2 · s^?1, at the agitation rate of 151 rpm in 24.5°C. The optimal physical process conditions for T. striata were obtained under the light intensity of 56 μmol photons · m^?2 · s^?1 in 25.5°C at the agitation rate of 151 rpm in 25.5°C, resulting in a specific growth rate of 0.226 d^?1. The predicted values were justified by the verification tests. Good agreement between the predicted values and the experimental values confirmed the validity of the models for the cultivation of microalgal strains. In this article, the noteworthy result was that temperature was a dominant factor in obtaining high chl‐a content for Nannochloropsis sp., whereas the growth of T. striata strongly depended on light exposure.     

Feeding,egg production,and egg hatching success of the copepods Acartia tonsa and Temora longicornis on diets of the toxic diatom Pseudo-nitzschia multiseries and the non-toxic diatom Pseudo-nitzschia pungens

In 1987, there was an episode of shellfish poisoning in Canada with human fatalities caused by the diatom Pseudo-nitzschia multiseries, which produced the toxin domoic acid. In order to examine whether domoic acid in this diatom serves as a grazing deterrent for copepods, we compared feeding rates, egg production rates, egg hatching success and mortality of the calanoid copepods Acartia tonsa and Temora longicornis feeding on unialgal diets of the toxic diatom P. multiseries and the similarly-sized non-toxic diatom Pseudo-nitzschia pungens. Copepods were collected in summers of 1994, 1995 and 1996 from Shediac Bay, New Brunswick, Canada, near Prince Edward Island, the site of the 1987 episode of domoic acid shellfish poisoning. Rates of ingestion of the toxic versus the non-toxic diatom by A. tonsa and T. longicornis were similar, with only one significantly different pair of values obtained in 1994, for which A. tonsa had a higher mean rate of ingestion of the toxic than the non-toxic diatom. Thus, domoic acid did not appear to retard grazing. Analyses of copepods with high performance liquid chromatography (HPLC) revealed that copepods accumulated domoic acid when feeding on P. multiseries. Egg production rates of copepods when feeding on P. multiseries and P. pungens were very low, ranging from 0 to 2.79 eggs female^–1 d^–1. There did not appear to be differential egg production or egg hatching success on diets of the toxic and non-toxic diatoms. Mortality of females on the toxic diet was low, ranging from 0 to 20%, with a mean of 13%, and there was no apparent difference between mortality of copepods feeding on toxic versus non-toxic diatoms. Egg hatching success on both diets, although based on few eggs, ranged between 22% and 76%, with a mean percentage hatching of 45%. Diets of the non-toxic diatom plus natural seawater assemblages supplemented with dissolved domoic acid, revealed similar rates and percentages when compared to previous experiments. In summary, none of the variables measured indicated adverse effects on copepods feeding on the toxic compared to the non-toxic diatom.     

EFFECTS OF Si:P CONCENTRATION RATIOS AND NUTRIENT LIMITATION ON THE CELLULAR COMPOSITION AND MORPHOLOGY OF ASTERIONELLA FORMOSA (BACILLARIOPHYCEAE)1

The freshwater diatom Asterionella formosa Haas. was grown in semicontinuous culture at 20°C under continuous cool-white fluorescent light of ca. 20 μEin · m^?2· s ·^?1 in a medium containing Si: P in various concentration ratios. The cell quotas of P and Si changed in relation to the available concentrations of P and Si at constant μ= 0.11 and 0.16 d^?1. Under Si-limitation, the P cell quota increased by over an order of magnitude as the influent [Si:P] decreased. The Si cell quota increased with increase in [Si] in the influent medium, and it increased as [P] increased at a specific [Si]. Under P-limitation, the P cell quotas were fairly constant and low; the Si cell quotas were relatively high and decreased slightly as influent [P] and [Si] increased. Asterionella stored up to 28 times more P and 2 times more Si than needed. The number of Asterionella cells per colony varied as a function of the influent [Si:P] and nutrient limitation being usually less than or equal to 6 when P-limited, and greater than 10 when Si-limited.     

AN ALTERNATE GROWTH PATTERN FOR LAMINARIA LONGICRURIS1,2

A population of Laminaria longicruris de la Pylaie was followed for a year at Bic Island, Quebec, Canada where nutrient levels in the seawater were elevated throughout the year. Tagged kelp were measured each month for growth and analyzed for alginic acid, laminaran, mannitol, carbon, nitrogen, and nitrate. Maximum growth (3.5 cm · d^?1) was observed in June, and minimal growth (0.18 cm · d^?1) from December to February, when ice cover limited light levels. No reserves of carbon or nitrate were formed. Laminaran levels remained below 2.7% dry weight while tissue nitrate did not exceed 0.75 μmol · g^?1 dry weight. Total carbon produced per plant was 40 g C · yr^?1. Nutrient availability enables the kelp to take advantage of summer light and temperature conditions to grow rapidly.     

EFFECT OF LIGHT AND TEMPERATURE INTERACTIONS ON GROWTH OF CRYPTOMONAS EROSA (CRYPTOPHYCEAE)1

Cryptomonas erosa Skuja, a planktonic alga, was grown in batch culture at different combinations of light intensity and temperature, under nutrient saturation. Growth was maximal (1.2 divisions · day^?1) at 23.5 C and 0.043 ly · min^?1, declining sharply with temperature (0.025 divisions-day^?1 at 1 C). With decreasing temperature, the cells showed both light saturation and inhibition at much reduced light intensities. At the same time the compensation light intensity for growth declined towards a minimum of slightly above 0.4 × 10^?4 ly · min^?1 (~1 ft-c) at 1 C or <0.1 ly · day^?1 (PAR). Cell division was more adversely affected by low temperature than carbon uptake, and the resulting excess production of photosynthate was both stored and excreted. Extreme storage of carbohydrates resulted in cell volumes and carbon content ca. 22 and 30 × greater, respectively, than the maxima observed for cells incubated in the dark, whereas, at growth inhibitory light levels, as much as 57% of the total assimilated carbon was excreted. A marked increase in cell pigment was observed at the lowest light levels (<10^?3 ly · min^?1), at high temperature. The growth response of C. erosa in culture provides insight into the abundance and distribution of cryptomonads and other small algal flagellates in nature.     

IMPORTANCE OF PHYSICAL VARIABLES ON THE SEASONAL DYNAMICS OF EPILITHIC ALGAE IN A HIGHLY SHADED CANYON STREAM1

The seasonal abundance of epilithic algae was correlated with major physico-chemical parameters in a first-order, heavily shaded stream in northern Arizona. Diatoms made up over 85%, by numerical abundance, of the epilithon community Light energy, water temperature, and stream discharge were most highly correlated with seasonal abundance of epilithic diatom taxa when analyzed with stepwise multiple regression. None of the chemical variables measured in the study (NO₃-N, O-PO₄, SiO₂, including PH) was found to be significantly correlated with the seasonal community structure of epilithic diatoms. Total diatom cell densities showed a significant negative correlation to stream bed light energy. Likewise, total diatom cell densities along a transect in the stream bed showed a negative correlation to current velocity during those months when base flow was low and stable, and current velocity was ≤25 cm·sec-¹. Most diatom taxa had highest cell densities at temperatures < 16°C and at daily mean stream bed light levels < 400 μE·m^?2·s^?1. Highest cell densities of green algae occurred at temperatures between 6–16°C and at daily mean stream bed light levels of > 400 μE·m^?2·s^?1. Blue-green algae (cyanobacteria) grew best at the highest recorded water temperatures and daily mean stream bed light energy (16–20°C and 900–1200 μE·m^?2·s^?1). Abrupt increases in NO₃-N coincided with a brief pulse of Nostoc pruniforme colonies during June, and leaf drop from Alnus oblongifolia during October.     

EFFECTS OF CELL DENSITY,TEMPERATURE, AND LIGHT INTENSITY ON GROWTH AND STALK PRODUCTION IN THE BIOFOULING DIATOM ACHNANTHES LONGIPES (BACILLARIOPHYCEAE)1

Achnanthes longipes Ag. is a marine stalk‐forming diatom that grows in dense biofilms. The effects of cell density, temperature, and light on growth and stalk production were examined in the laboratory to determine how they affected the ability of this diatom to form a biofilm. Stalk production abruptly increased when A. longipes was cultured at a density of 5.4 × 10³ cells·mL ^{? 1} 1 Received 23 February 2002. Accepted 22 July 2002.
, with a lag before stalk production occurring in cultures initiated at lower densities. Growth occurred at all temperatures from 8 to 32° C, with maximum growth at 26° C. Growth rate was light saturated at 60 μmol photons·m ^{? 2}·s ^{? 1} 1 Received 23 February 2002. Accepted 22 July 2002.
. Stalk production was determined as the proportion of cells producing stalks and stalk length in response to various temperatures and light intensities at high (5000 cells·mL ^{? 1} 1 Received 23 February 2002. Accepted 22 July 2002.
) and low (500 cells·mL ^{? 1} 1 Received 23 February 2002. Accepted 22 July 2002.
) densities. More cells formed stalks at high density, with no difference in stalk length. The proportion of cells producing stalks was maximal at 20° C, with little change at 17–32° C. Stalk length was at a maximum between 14 and 26° C. Stalk production showed little change in response to varying light intensity. The results of an earlier investigation on the effects of bromide concentration on stalk formation were expressed as the proportion of cells forming stalks and the lengths of the stalks. Both measures of stalk production varied with bromide concentration, with maximum values at 30 mM bromide. The increased stalk production at higher densities may be a means of elevating cells above the substrate to avoid competition in the dense biofilm.     

CONFIRMATION OF DOMOIC ACID PRODUCTION BY PSEUDO‐NITZSCHIA AUSTRALIS (BACILLARIOPHYCEAE) ISOLATED FROM IRISH WATERS1

A nonaxenic isolate of the potentially toxic diatom Pseudo‐nitzschia australis (Frenguelli) from Irish waters was tested in two separate batch culture experiments. When grown under a low irradiance (～12 μmol photons·m ^{? 2}·s ^{? 1} 1 Received 20 March 2001. Accepted 21 August 2002.
; 16:8‐h light:dark cycle) for up to 40 days, the culture produced only trace amounts of the neurotoxin domoic acid (DA) during late stationary phase. Growth at a higher irradiance (～115 μmol photons·m ^{? 2}·s ^{? 1} 1 Received 20 March 2001. Accepted 21 August 2002.
; 12:12‐h light:dark cycle) resulted in DA production starting during late exponential phase and reaching a maximum concentration of 26 pg DA·cell ^{? 1} 1 Received 20 March 2001. Accepted 21 August 2002.
during late stationary phase. Liquid chromatography coupled to mass spectrometry was used to confirm the identity of DA in the culture. Irradiance and photoperiod could be important factors that contribute directly or indirectly to the control of DA production in P. australis. This is the first record of a DA‐producing diatom in Irish waters, and results indicate P. australis may have been the source of DA that has recently contaminated shellfisheries in this area.     

EFFECTS OF LIGHT AND SUBSTRATE ON THE BENTHIC DIATOMS IN AN OLIGOTROPHIC LAKE: A COMPARISON BETWEEN NATURAL AND ARTIFICIAL SUBSTRATES1

Benthic diatoms form a particularly important community in oligotrophic lakes, but factors influencing their distribution are not well known. This study reports the depth distribution of living motile and total diatoms (living plus dead diatoms) on both natural (from sand to fine organic mud) and artificial substrates in an oligotrophic lake. On artificial substrates, motile diatom densities peaked in abundance (24–30 cells · mm^?2) between 0.6 and 1.9 m depth; on natural sediment surfaces, motile diatoms were generally more numerous and peaked in abundance (925 cells · mm^?2) at 1.3 m depth. Total diatom densities on artificial substrates were highest (1260 valves · mm^?2) at 0.6 m depth, with very low values below 3 m depth; on natural sediment surfaces, total diatom abundances were generally much higher (21600 valves · mm^?2) at 3 m depth and declined gradually with depth. Significant relationships were found between light and diatom densities on the artificial substrate. Ordination analysis indicated that substrate type significantly correlated with the variation of diatom composition on artificial and natural substrates. Our results suggest that in oligotrophic lakes, light influences benthic diatom abundance, whereas substrate type has more influence on benthic diatom composition.     

Light History Influences the Response of Fluvial Biofilms to Zn Exposure

Fluvial biofilms are subject to multistress situations in natural ecosystems, such as the co‐occurrence of light intensity changes and metal toxicity. However, studies simultaneously addressing both factors are rare. This study evaluated in microcosm conditions the relationship between short‐term light intensity changes and Zn toxicity on fluvial biofilms with long‐term photoacclimation to different light conditions. Biofilms that had long‐term photoacclimation to 25 μmol photons · m^?2 · s^?1 (low light [LL] biofilms), 100 μmol photons · m^?2 · s^?1 (medium light [ML] biofilms), and 500 μmol photons · m^?2 · s^?1 (high light [HL] biofilms) were characterized by different structural (Chlorophyll‐a [Chl‐a], total biomass‐AFDW, EPS, algal groups, and diatom taxonomy) and physiological attributes (ETR‐I curves and photosynthetic pigments). HL biofilms showed higher light saturation intensity and a higher production of xanthophylls than LL biofilms. In contrast, LL biofilms had many structural differences; a higher proportion of diatoms and lower AFDW and EPS contents than ML and HL biofilms. A clear effect of light intensity changes on Zn toxicity was also demonstrated. Zn toxicity was enhanced when a sudden increase in light intensity also occurred, mainly with LL biofilms, causing higher inhibition of both the Φ′_PSII and the Φ_PSII. A decoupling of NPQ from de‐epoxidation reaction (DR) processes was also observed, indicating substantial damage to photoprotective mechanisms functioning in biofilms (i.e., xanthophyll cycle of diatoms) due to Zn toxicity. This study highlights the need to take into account environmental stress (e.g., light intensity changes) to better assess the environmental risks of chemicals (e.g., metals).     

LIGHT,TEMPERATURE AND PHOTOPERIOD AS FACTORS CONTROLLING REPRODUCTION IN ULOTHRIX ZONATA (ULVOPHYCEAE)1

In many lakes in the northern United States and Canada the filamentous green alga Ulothrix zonata (Weber and Mohr) Kütz grows abundantly in early spring in shallow waters. Asexual reproduction occurs by formation of quadriflagellate zoospores which disrupt, the integrity of the cells upon release causing the filament to disintegrate. Study of the effects of 100 different combinations of irradiance, temperature and photoperiod revealed that zoospore formation is favored by high temperatures near 20°C, high light levels of 520 μE·m^?2·s^?1 and photoperiods of either short day (8:16 h light-dark) or long day cycles (16:8 h light-dark). Zoospore formation is minimal under conditions of low temperature (5°C), low irradiance (32.5 μE·m^?2·s^?1) and neutral day-lengths (12:12 h light-dark). These observations explain the decline in U. zonata biomass when water temperatures rise above 10° C. The combined effect of rising water temperatures and increasing daylengths causes progressively more filaments to switch from vegetable growth to zoospore production resulting in an increasing loss of biomass.     

Differential effects of changes in spectral irradiance on photoacclimation,primary productivity and growth in Rhodomonas salina (Cryptophyceae) and Skeletonema costatum (Bacillariophyceae) in simulated blackwater environments

The underwater light field in blackwater environments is strongly skewed toward the red end of the electromagnetic spectrum due to blue light absorption by colored dissolved organic matter (CDOM). Exposure of phytoplankton to full spectrum irradiance occurs only when cells are mixed up to the surface. We studied the potential effects of mixing‐induced changes in spectral irradiance on photoacclimation, primary productivity and growth in cultures of the cryptophyte Rhodomonas salina and the diatom Skeletonema costatum. We found that these taxa have very different photoacclimation strategies. While S. costatum showed classical complementary chromatic adaption, R. salina showed inverse chromatic adaptation, a strategy previously unknown in the cryptophytes. Transfer of R. salina to periodic full spectrum light (PFSL) significantly enhanced growth rate (μ) by 1.8 times and primary productivity from 0.88 to 1.35 mg C · (mg Chl^?1) · h^?1. Overall, R. salina was less dependent on PFSL than was S. costatum, showing higher μ and net primary productivity rates. In the high‐CDOM simulation, carbon metabolism of the diatom was impaired, leading to suppression of growth rate, short‐term ¹⁴C uptake and net primary production. Upon transfer to PFSL, μ of the diatom increased by up to 3‐fold and carbon fixation from 2.4 to 6.0 mg C · (mg Chl^?1) · h^?1. Thus, a lack of PFSL differentially impairs primarily CO₂‐fixation and/or carbon metabolism, which, in turn, may determine which phytoplankton dominate the community in blackwater habitats and may therefore influence the structure and function of these ecosystems.     

THE INFLUENCE OF SUBMERGED MACROPHYTES ON SEDIMENTARY DIATOM ASSEMBLAGES1

Submerged macrophytes are a central component of lake ecosystems; however, little is known regarding their long‐term response to environmental change. We have examined the potential of diatoms as indicators of past macrophyte biomass. We first sampled periphyton to determine whether habitat was a predictor of diatom assemblage. We then sampled 41 lakes in Quebec, Canada, to evaluate whether whole‐lake submerged macrophyte biomass (BiomEpiV) influenced surface sediment diatom assemblages. A multivariate regression tree (MRT) was used to construct a semiquantitative model to reconstruct past macrophyte biomass. We determined that periphytic diatom assemblages on macrophytes were significantly different from those on wood and rocks (ANOSIM R = 0.63, P < 0.01). A redundancy analysis (RDA) of the 41‐lake data set identified BiomEpiV as a significant (P < 0.05) variable in structuring sedimentary diatom assemblages. The MRT analysis classified the lakes into three groups. These groups were (A) high‐macrophyte, nutrient‐limited lakes (BiomEpiV ≥525 μg · L^?1; total phosphorus [TP] <35 μg · L^?1; 23 lakes); (B) low‐macrophyte, nutrient‐limited lakes (BiomEpiV <525 μg · L^?1; TP <35 μg · L^?1; 12 lakes); and (C) eutrophic lakes (TP ≥35 μg · L^?1; six lakes). A semiquantitative model correctly predicted the MRT group of the lake 71% of the time (P < 0.001). These results suggest that submerged macrophytes have a significant influence on diatom community structure and that sedimentary diatom assemblages can be used to infer past macrophyte abundance.     

THE PERFORMANCE OF SPRAY‐IRRIGATED ULVA LACTUCA (ULVOPHYCEAE,CHLOROPHYTA) AS A CROP AND AS A BIOFILTER OF FISHPOND EFFLUENTS1

The seaweed Ulva lactuca L. was spray cultured by mariculture effluents in a mattress‐like layer, held in air on slanted boards by plastic netting. Air‐agitated seaweed suspension tanks were the reference. Growth rate, yield, and ammonia‐N removal rate were 11.8% · d^?1, 171 g fresh weight (fwt) · m^?2 · d^?1, and 5 g N · m^?2 · d^?1, respectively, by the spray‐cultured U. lactuca, and 16.9% · d^?1, 283 g fwt · m^?2 · d^?1, and 7 g N · m^?2 · d^?1, respectively, by the tank U. lactuca. Biomass protein content was similar in both treatments. Dissolved oxygen in the fishpond effluent water was raised by >3 mg · L^?1 and pH by up to half a unit, upon passage through both culture systems. The data suggest that spray‐irrigation culture of U. lactuca in this simple green‐mattress‐like system supplies the seaweed all it needs to grow and biofilter at rates close to those in standard air‐agitated tank culture.     

CHANGES IN DOMOIC ACID PRODUCTION AND CELLULAR CHEMICAL COMPOSITION OF THE TOXIGENIC DIATOM PSEUDO-NITZSCHIA MULTISERIES UNDER PHOSPHATE LIMITATION1

Production of domoic acid (DA), a neurotoxin, by the diatom Pseudo-nitzschia multiseries (previously Nitzschia pungens f. multiseries) Hasle and its cellular chemical composition were studied in phosphate-limited chemostat continuous cultures and in subsequent batch cultures. Under steady-state chemostat conditions, DA production increased from 0.01 to 0.26 pg DA · cell^?1· d^?1 as the growth rate decreased. When the nutrient supply was discontinued (to produce a batch culture), DA production was enhanced by a factor of ca. 3. DA production was temporarily suspended upon addition of phosphate to the batch cultures but resumed 1 d later at a higher rate coincident with the decline of phosphate uptake. In both steady-state continuous culture and batch culture, more DA was produced when alkaline phosphatase activity (APA) was high. The association of high DA production with high levels of APA and high cellular N:P ratios strongly suggests that phosphate limitation enhances DA production. Also, DA production was high when other primary metabolism (e.g. uptake of carbon, nitrogen, phosphorus and silicon, and cell division) was low, but chlorophyll a and adenosine triphosphate were generally high. This suggests that the synthesis of DA requires a substantial amount of biogenic energy.     

Effects of light and temperature on the growth of Takayama helix (Dinophyceae): mixotrophy as a survival strategy against photoinhibition

Takayama helix is a mixotrophic dinoflagellate that can feed on diverse algal prey. We explored the effects of light intensity and water temperature, two important physical factors, on its autotrophic and mixotrophic growth rates when fed on Alexandrium minutum CCMP1888. Both the autotrophic and mixotrophic growth rates and ingestion rates of T. helix on A. minutum were significantly affected by photon flux density. Positive growth rates of T. helix at 6–58 μmol photons · m^?2 · s^?1 were observed in both the autotrophic (maximum rate = 0.2 · d^?1) and mixotrophic modes (0.4 · d^?1). Of course, it did not grow both autotrophically and mixotrophically in complete darkness. At ≥247 μmol photons · m^?2 · s^?1, the autotrophic growth rates were negative (i.e., photoinhibition), but mixotrophy turned these negative rates to positive. Both autotrophic and mixotrophic growth and ingestion rates were significantly affected by water temperature. Under both autotrophic and mixotrophic conditions, it grew at 15–28°C, but not at ≤10 or 30°C. Therefore, both light intensity and temperature are critical factors affecting the survival and growth of T. helix.     

Nitrogen Utilization and Toxin Production by Two Diatoms of the Pseudo‐nitzschia pseudodelicatissima Complex: P. cuspidata and P. fryxelliana

The toxigenic diatom Pseudo‐nitzschia cuspidata, isolated from the U.S. Pacific Northwest, was examined in unialgal batch cultures to evaluate domoic acid (DA) toxicity and growth as a function of light, N substrate, and growth phase. Experiments conducted at saturating (120 μmol photons · m^?2 · s^?1) and subsaturating (40 μmol photons · m^?2 · s^?1) photosynthetic photon flux density (PPFD), demonstrate that P. cuspidata grows significantly faster at the higher PPFD on all three N substrates tested [nitrate (NO₃^?), ammonium (NH₄⁺), and urea], but neither cellular toxicity nor exponential growth rates were strongly associated with one N source over the other at high PPFD. However, at the lower PPFD, the exponential growth rates were approximately halved, and the cells were significantly more toxic regardless of N substrate. Urea supported significantly faster growth rates, and cellular toxicity varied as a function of N substrate with NO₃^?‐supported cells being significantly more toxic than both NH₄⁺‐ and urea‐supported cells at the low PPFD. Kinetic uptake parameters were determined for another member of the P. pseudodelicatissima complex, P. fryxelliana. After growth of these cells on NO₃^? they exhibited maximum specific uptake rates (V_max) of 22.7, 29.9, 8.98 × 10^?3 · h^?1, half‐saturation constants (K_s) of 1.34, 2.14, 0.28 μg‐at N · L^?1, and affinity values (α) of 17.0, 14.7, 32.5 × 10^?3 · h^?1/(μg‐at N · L^?1) for NO₃^?, NH₄⁺ and urea, respectively. These labo‐ratory results demonstrate the capability of P. cuspidata to grow and produce DA on both oxidized and reduced N substrates during both exponential and stationary growth phases, and the uptake kinetic results for the pseudo‐cryptic species, P. fryxelliana suggest that reduced N sources from coastal runoff could be important for maintenance of these small pennate diatoms in U.S. west coast blooms, especially during times of low ambient N concentrations.