Influence of daylight surface aggregation behavior on nutrient cycling during a Karenia brevis (Davis) G. Hansen & Ø. Moestrup bloom: Migration to the surface as a nutrient acquisition strategy

The toxic HAB dinoflagellate Karenia brevis (Davis) G. Hansen & Ø. Moestrup (formerly Gymnodinium breve) exhibits a migratory pattern atypical of dinoflagellates: cells concentrate in a narrow (∼0–5 cm) band at the water surface during daylight hours due to phototactic and negative geotactic responses, then disperse downward at night via non-tactic, random swimming. The hypothesis that this daylight surface aggregation behavior significantly influences bacterial and algal productivity and nutrient cycling within blooms was tested during a large, high biomass (chlorophyll a >19 μg L⁻¹) K. brevis bloom in October of 2001 by examining the effects of this surface layer aggregation on inorganic and organic nutrient concentrations, cellular nitrogen uptake, primary and bacterial productivity and the stable isotopic signature (δ¹⁵N, δ¹³C) of particulate material. During daylight hours, concentrations of K. brevis and chlorophyll a in the 0–5 cm surface layer were enhanced by 131% (±241%) and 32.1% (±86.1%) respectively compared with an integrated water sample collection over a 0–1 m depth. Inorganic (NH₄, NO₃₊₂, PO₄, SiO₄) and organic (DOP, DON) nutrient concentrations were also elevated within the surface layer as was both bacterial and primary productivity. Uptake of nitrogen (NH₄⁺, NO₃⁻, urea, dissolved primary amines, glutamine and alanine) compounds by K. brevis was greatest in the surface layer for all compounds tested, with the greatest enhancement evident in urea uptake rates, from 0.08 × 10⁻⁵ ng N K. brevis cell⁻¹ h⁻¹ to 3.1 × 10⁻⁵ ng N K. brevis cell⁻¹ h⁻¹. These data suggests that this surface aggregation layer is not only an area of concentrated cells within K. brevis blooms, but also an area of increased biological activity and nutrient cycling, especially of nitrogen. Additionally, the classic dinoflagellate migration paradigm of a downward migration for access to elevated NO₃⁻ concentrations during the dark period may not apply to certain dinoflagellates such as K. brevis in oligotrophic nearshore areas with no significant nitricline. For these dinoflagellates, concentration within a narrow surface layer in blooms during daylight hours may enhance nutrient supply through biological cycling and photochemical nutrient regeneration.     

Temporal and spatial variability in nitrogen uptake kinetics during harmful dinoflagellate blooms in the East China Sea

Temporal and spatial variability in the kinetic parameters of uptake of nitrate (NO₃⁻), ammonium (NH₄⁺), urea, and glycine was measured during dinoflagellate blooms in Changjiang River estuary and East China Sea coast, 2005. Karenia mikimotoi was the dominant species in the early stage of the blooms and was succeeded by Prorocentrum donghaiense. The uptake of nitrogen (N) was determined using ¹⁵N tracer techniques. The results of comparison kinetic parameters with ambient nutrients confirmed that different N forms were preferentially taken up during different stages of the bloom. NO₃⁻ (V_max 0.044 h⁻¹; K_s 60.8 μM-N) was an important N source before it was depleted. NH₄⁺ (V_max 0.049 h⁻¹; K_s 2.15 μM-N) was generally the preferred N. Between the 2 organic N sources, urea was more preferred when K. mikimotoi dominated the bloom (V_max 0.020 h⁻¹; K_s 1.35 μM-N) and glycine, considered as a dominant amino acid, was more preferred when P. donghaiense dominated the bloom (V_max 0.025 h⁻¹; K_s 1.76 μM-N). The change of N uptake preference by the bloom-forming algae was also related to the variation in ambient N concentrations.     

Nitrogen uptake kinetics of Prymnesium parvum (Haptophyte)

The uptake rates of different nitrogen (N) forms (NO₃⁻, urea, and the amino acids glycine and glutamic acid) by N-deficient, laboratory-grown cells of the mixotrophic haptophyte, Prymnesium parvum, were measured and the preference by the cells for the different forms determined. Cellular N uptake rates (ρ_cell, fmol N cell⁻¹ h⁻¹) were measured using ¹⁵N-labeled N substrates. P. parvum showed high preference for the tested amino acids, in particular glutamic acid, over urea and NO₃⁻ under the culture nutrient conditions. However, extrapolating these rates to Baltic Seawater summer conditions, P. parvum would be expected to show higher uptake rates of NO₃⁻ and the amino acids relative to urea because of the difference in average concentrations of these substrates. A high uptake rate of glutamic acid at low substrate concentrations suggests that this substrate is likely used through extracellular enzymes. Nitrate, urea and glycine, on the other hand, showed a non-saturating uptake over the tested substrate concentration (1–40 μM-N for NO₃⁻ and urea, 0.5–10 μM-N for glycine), indicating slower membrane-transport rates for these substrates.     

The effects of NH4+ and NO3− on growth,resource allocation and nitrogen uptake kinetics of Phragmites australis and Glyceria maxima

The effects of NH₄⁺ or NO₃⁻ on growth, resource allocation and nitrogen (N) uptake kinetics of two common helophytes Phragmites australis (Cav.) Trin. ex Steudel and Glyceria maxima (Hartm.) Holmb. were studied in semi steady-state hydroponic cultures. At a steady-state nitrogen availability of 34 μM the growth rate of Phragmites was not affected by the N form (mean RGR = 35.4 mg g⁻¹ d⁻¹), whereas the growth rate of Glyceria was 16% higher in NH₄⁺-N cultures than in NO₃⁻-N cultures (mean = 66.7 and 57.4 mg g⁻¹ d⁻¹ of NH₄⁺ and NO₃⁻ treated plants, respectively). Phragmites and Glyceria had higher S/R ratio in NH₄⁺ cultures than in NO₃⁻ cultures, 123.5 and 129.7%, respectively.Species differed in the nitrogen utilisation. In Glyceria, the relative tissue N content was higher than in Phragmites and was increased in NH₄⁺ treated plants by 16%. The tissue NH₄⁺ concentration (mean = 1.6 μmol g fresh wt⁻¹) was not affected by N treatment, whereas NO₃⁻ contents were higher in NO₃⁻ (mean = 1.5 μmol g fresh wt⁻¹) than in NH₄⁺ (mean = 0.4 μmol g fresh wt⁻¹) treated plants. In Phragmites, NH₄⁺ (mean = 1.6 μmol g fresh wt⁻¹) and NO₃⁻ (mean = 0.2 μmol g fresh wt⁻¹) contents were not affected by the N regime. Species did not differ in NH₄⁺ (mean = 56.5 μmol g⁻¹ root dry wt h⁻¹) and NO₃⁻ (mean = 34.5 μmol g⁻¹ root dry wt h⁻¹) maximum uptake rates (V_max), and V_max for NH₄⁺ uptake was not affected by N treatment. The uptake rate of NO₃⁻ was low in NH₄⁺ treated plants, and an induction phase for NO₃⁻ was observed in NH₄⁺ treated Phragmites but not in Glyceria. Phragmites had low K_m (mean = 4.5 μM) and high affinity (10.3 l g⁻¹ root dry wt h⁻¹) for both ions compared to Glyceria (K_m = 6.3 μM, affinity = 8.0 l g⁻¹ root dry wt h⁻¹). The results showed different plasticity of Phragmites and Glyceria toward N source. The positive response to NH₄⁺-N source may participates in the observed success of Glyceria at NH₄⁺ rich sites, although other factors have to be considered. Higher plasticity of Phragmites toward low nutrient availability may favour this species at oligotrophic sites.     

Contribution of diazotrophy to nitrogen inputs supporting Karenia brevis blooms in the Gulf of Mexico

Blooms of Karenia brevis plague the West Florida Shelf (WFS) region in the Gulf of Mexico (GOM) where they exert harmful effects on aquatic biota and humans. Because productivity on the WFS is N limited, new N inputs into the region are thought to trigger blooms of K. brevis. Here we examine the potential for new N inputs via N₂ fixation by Trichodesmium and other diazotrophic plankton to contribute to the N demand of K. brevis. Because of possible methodological biases, we also compared N₂ fixation rates by cultured Trichodesmium using the ¹⁵N₂ bubble addition method and the ¹⁵N₂ saturated seawater. Both methods yielded identical results in 12 and 24 h incubations; however, there was more variability in rate estimates made using the bubble addition method. Pelagic N₂ fixation rates by other planktonic diazotrophs ranged from 0 to 13.6 nmol N L⁻¹ d⁻¹, comparable to or higher than rates observed in oligotrophic gyres. These rates should be considered conservative estimates because they were made using the bubble addition method. Integrating over our study area, we estimate that new inputs of N to the WFS via N₂ fixation are on the order of 0.011 Tmol N annually. Further, we measured directly the trophic transfer of recently fixed N₂ to co-occurring plankton that included K. brevis and found that up to 47% of N₂ fixed was transferred to non-diazotrophic plankton even in short (<6 h) incubations where N₂ fixation was likely underestimated.     

Uptake and inhibition kinetics of nitrogen in Microcystis aeruginosa: Results from cultures and field assemblages collected in the San Francisco Bay Delta,CA

The nitrogen (N) uptake kinetic parameters for Microcystis field assemblages collected from the San Francisco Bay Delta (Delta) in 2012 and non-toxic and toxic laboratory culture strains of M. aeruginosa were assessed. The ¹⁵N tracer technique was used to investigate uptake of ammonium (NH₄⁺), nitrate (NO₃⁻), urea and glutamic acid over short-term incubations (0.5–1 h), and to study inhibition of NO₃⁻, NH₄⁺ and urea uptake by NH₄⁺, NO₃⁻ and NH₄⁺, respectively. This study demonstrates that Delta Microcystis can utilize different forms of inorganic and organic N, with the greatest capacity for NH₄⁺ uptake and the least for glutamic acid uptake, although N uptake did not always follow the classic Michaelis–Menten hyperbolic relationship at substrate concentrations up to 67 μmol N L⁻¹. Current ambient N concentrations in the Delta may be at sub-saturating levels for N uptake, indicating that if N loading (especially NH₄⁺) were to increase, Delta Microcystis assemblages have the potential for increased N uptake rates. Delta Microcystis had the highest specific affinity, α, for NH₄⁺ and the lowest for NO₃⁻. In culture, N uptake by non-toxic and toxic M. aeruginosa strains was much higher than from the field, but followed similar N utilization trends to those in the field. Neither strain showed severe inhibition of NO₃⁻ uptake by NH₄⁺ or inhibition of NH₄⁺ uptake on NO₃⁻, but both strains showed some inhibition of urea uptake by NH₄⁺.     

Morphology and bloom dynamics of Cochlodinium geminatum (Schütt) Schütt in the Pearl River Estuary,South China Sea

An unarmored dinoflagellate bloom of Cochlodinium geminatum (Schütt) Schütt has been identified in the Pearl River Estuary, South China Sea during the severe dry season, from late October to early November, 2009, when temperature and salinity ranged between 20.0–27.2 °C and 10.6–33.4, respectively. Light and scanning electron microscopy were used to identify the characteristics of C. geminatum and provided the clear morphological structure for this species. The organism was primarily found in chains of two cells or single cell, and no longer chains were observed. Cells were irregularly spherical or slightly dorso-ventrally, with size ranged between 28 and 36 μm and longer than wide. A large nucleus in the center with numerous golden chloroplasts was present, and the cingulum made 1.5 turns around the cell. The concentration of C. geminatum ranged from 10² to greater than 10⁷ cells l⁻¹ during the bloom period. Nutrient concentration ranges during the bloom were 1.29–81.00 μM NO₃, 0.14–12.14 μM NO₂, 0.21–6.29 μM NH₄, 0.23–6.26 μM PO₄ and 3.29–171.43 μM SiO₃, respectively. Total biomass expressed in terms of chlorophyll a ranged from 2.44 to 135.45 μg l⁻¹, with an average 19.9 μg l⁻¹ in surface water throughout the PRE. Two main clusters corresponding to the water sectors were defined with multivariate analysis (cluster and nMDS). Based on the composition and abundance of phytoplankton, spatial variations were observed at a significant level (ANOSIM, R = 0.44, P < 0.01). Although the pairwise correlation analysis detected no significant effect of any single environmental variable on the abundance of C. geminatum, the multivariate analysis (BIO-ENV) between biotic and abiotic variables resulted in the best variables combination with all measured factors involved (temperature, salinity, turbidity, NO₃, NO₂, NH₄, PO₄ and SiO₃) which showed a combined effect during the bloom of C. geminatum in the Pearl River Estuary (ρ_w = 0.477).     

Detection of Karenia brevis blooms on the west Florida shelf using in situ backscattering and fluorescence data

Using shipboard data collected from the central west Florida shelf (WFS) between 2000 and 2001, an optical classification algorithm was developed to differentiate toxic Karenia brevis blooms (>10⁴ cells l⁻¹) from other waters (including non-blooms and blooms of other phytoplankton species). The identification of K. brevis blooms is based on two criteria: (1) chlorophyll a concentration ≥1.5 mg m⁻³ and (2) chlorophyll-specific particulate backscattering at 550 nm ≤ 0.0045 m² mg⁻¹. The classification criteria yielded an overall accuracy of 99% in identifying both K. brevis blooms and other waters from 194 cruise stations. The algorithm was validated using an independent dataset collected from both the central and south WFS between 2005 and 2006. After excluding data from estuarine and post-hurricane turbid waters, an overall accuracy of 94% was achieved with 86% of all K. brevis bloom data points identified successfully. Satisfactory algorithm performance (88% overall accuracy) was also achieved when using underway chlorophyll fluorescence and backscattering data collected during a repeated alongshore transect between Tampa Bay and Florida Bay in 2005 and 2006. These results suggest that it may be possible to use presently available, commercial optical backscattering instrumentation on autonomous platforms (e.g. moorings, gliders, and AUVs) for rapid and timely detection and monitoring of K. brevis blooms on the WFS.     

Uptake of dissolved inorganic and organic nitrogen by the benthic toxic dinoflagellate Ostreopsis cf. ovata

Environmental factors that shape dynamics of benthic toxic blooms are largely unknown. In particular, for the toxic dinoflagellate Ostreopsis cf. ovata, the importance of the availability of nutrients and the contribution of the inorganic and organic pools to growth need to be quantified in marine coastal environments. The present study aimed at characterizing N-uptake of dissolved inorganic and organic sources by O. cf. ovata cells, using the ¹⁵N-labelling technique. Experiments were conducted taking into account potential interactions between nutrient uptake systems as well as variations with the diel cycle. Uptake abilities of O. cf. ovata were parameterized for ammonium (NH₄⁺), nitrate (NO₃⁻) and N-urea, from the estimation of kinetic and inhibition parameters. In the range of 0 to 10 μmol N L⁻¹, kinetic curves showed a clear preference pattern following the ranking NH₄⁺ > NO₃⁻ > N-urea, where the preferential uptake of NH₄⁺ relative to NO₃⁻ was accentuated by an inhibitory effect of NH₄⁺ concentration on NO₃⁻ uptake capabilities. Conversely, under high nutrient concentrations, the preference for NH₄⁺ relative to NO₃⁻ was largely reduced, probably because of the existence of a low-affinity high capacity inducible NO₃⁻ uptake system. Ability to take up nutrients in darkness could not be defined as a competitive advantage for O. cf. ovata. Species competitiveness can also be defined from nutrient uptake kinetic parameters. A strong affinity for NH₄⁺ was observed for O. cf. ovata cells that may partly explain the success of this toxic species during the summer season in the Bay of Villefranche-sur-mer (France).     

Vertical migration of Karenia brevis in the northeastern Gulf of Mexico observed from glider measurements

The toxic marine dinoflagellate, Karenia brevis (the species responsible for most of red tides or harmful algal blooms in the Gulf of Mexico), is known to be able to swim vertically to adapt to the light and nutrient environments, nearly all such observations have been made through controlled experiments using cultures. Here, using continuous 3-dimensional measurements by an ocean glider across a K. brevis bloom in the northeastern Gulf of Mexico between 1 and 8 August 2014, we show the vertical migration behavior of K. brevis. Within the bloom where K. brevis concentration is between 100,000 and 1,000,000 cells L⁻¹, the stratified water shows a two-layer system with the depth of pycnocline ranging between 14–20 m and salinity and temperature in the surface layer being <34.8 and >28 °C, respectively. The bottom layer shows the salinity of >36 and temperature of <26 °C. The low salinity is apparently due to coastal runoff, as the top layer also shows high amount of colored dissolved organic matter (CDOM). Within the top layer, chlorophyll-a fluorescence shows clear diel changes in the vertical structure, an indication of K. brevis vertical migration at a mean speed of 0.5–1 m h⁻¹. The upward migration appears to start at sunrise at a depth of 8–10 m, while the downward migration appears to start at sunset (or when surface light approaches 0) at a depth of ∼2 m. These vertical migrations are believed to be a result of the need of K. brevis cells for light and nutrients in a stable, stratified, and CDOM-rich environment.     

Comparative diel oxygen cycles preceding and during a Karenia bloom in Sarasota Bay,Florida, USA

The diel change in dissolved oxygen concentrations were recorded with an automated incubator containing a pulsed oxygen sensor in Sarasota Bay, Florida. The deployments occurred during a ‘pre-bloom’ period in May to June 2006, and during a harmful algal bloom dominated by Karenia brevis in September 2006. The diurnal (daylight) increase in dissolved oxygen concentrations varied from 16 to 104 μmol O₂ l⁻¹ with the corresponding nocturnal decrease in oxygen varying from 16 to 77 μmol O₂ l⁻¹. Nocturnal respiration consumed 42–113% of the diurnal net oxygen production with the minimum and maximum during the pre-bloom period. Hourly production rates closely followed fluctuations in irradiance with maximum rates in the late morning. Hourly oxygen utilization rates (community respiration) at night were highest during the first few hours after sunset.     

A nutrient biotic index (NBI) for use with benthic macroinvertebrate communities

Aquatic macroinvertebrates have been among the principal biological communities used for freshwater monitoring and assessment for several decades, but macroinvertebrate biomonitoring has not incorporated nutrient measures into assessment strategies. Two nutrient biotic indices were developed for benthic macroinvertebrate communities, one for total phosphorus (NBI-P), and one for nitrate (NBI-N). Weighted averaging was used to assess the distributions of 164 macroinvertebrate taxa across TP and NO₃⁻ gradients and to establish nutrient optima and subsequent nutrient tolerance values. Both the NBI-P and NBI-N were correlated with increasing mean TP and NO₃⁻ values (r = 0.68 and r = 0.57, respectively, p < 0.0001). A three-tiered scale of eutrophication for TP and NO₃⁻ (oligotrophic: ≤0.0175 mg/l TP, ≤0.24 mg/l NO₃⁻, mesotrophic: >0.0175 to ≤0.065 mg/l TP, >0.24 to ≤0.98 mg/l NO₃⁻, eutrophic: >0.065 mg/l TP, >0.98 mg/l NO₃⁻) was also established through cluster analysis of invertebrate communities using Bray–Curtis (quantitative) similarity. Significant differences (p < 0.0001) were detected between median NBI-P and NBI-N scores among the three trophic states. Therefore, the nutrient biotic indices (NBIs) appear to accurately reflect changes in stream trophic state. Multimetric water quality assessments were also used to identify thresholds of impairment among the three trophic states. Hodges-Lehman estimation indicated that the greatest change in assessment results occurred between the mesotrophic and eutrophic states. The eutrophic state also represented the highest percentage of overall impairment. Therefore, the suggested threshold for nutrient impairment is the boundary between mesotrophic and eutrophic (0.065 mg/l TP and 0.98 mg/l NO₃⁻). The corresponding NBI-P score (6.1) and NBI-N score (6.0) for this threshold incorporate predictive capabilities into the NBIs. The NBI and index score thresholds of impairment will provide monitoring programs with a robust measure of stream nutrient status and serve as a useful tool in enforcing regional nutrient criteria.     

Effect of NH4+/NO3− availability on nitrate reductase activity and nitrogen accumulation in wetland helophytes Phragmites australis and Glyceria maxima

The effect of NH₄⁺/NO₃⁻ availability on nitrate reductase (NR) activity in Phragmites australis and Glyceria maxima was studied in sand and water cultures with the goal to characterise the relationship between NR activity and NO₃⁻ availability in the rhizosphere and to describe the extent to which NH₄⁺ suppresses the utilization of NO₃⁻ in wetland plants.The NR activity data showed that both wetland helophytes are able to utilize NO₃⁻. This finding was further supported by sufficient growth observed under the strict NO₃⁻ nutrition. The distribution of NR activity within plant tissues differed between species. Phragmites was proved to be preferential leaf NO₃⁻ reducer with high NR activity in leaves (NR_max > 6.5 μmol NO₂⁻ g dry wt⁻¹ h⁻¹) under all N treatments, and therefore Phragmites seems to be good indicator of NO₃⁻ availability in flooded sediment. In the case of Glyceria the contribution of roots to plant NO₃⁻ reduction was higher, especially in sand culture. Glyceria also tended to accumulate NO₃⁻ in non-reduced form, showing generally lower leaf NR activity levels. Thus, the NR activity does not necessarily correspond with plant ability to take up NO₃⁻ and grow under NO₃⁻-N source. Moreover, the species differed significantly in the content of compounds interfering with NR activity estimation. Glyceria, but not Phragmites, contained cyanogenic glycosides releasing cyanide, the potent NR inhibitor. It clearly shows that the use of NR activity as a marker of NO₃⁻ utilization in individual plant species is impossible without the precise method optimisation.     

Monitoring,management, and mitigation of Karenia blooms in the eastern Gulf of Mexico

Annual blooms of the toxic dinoflagellate Karenia brevis in the eastern Gulf of Mexico represent one of the most predictable global harmful algal bloom (HAB) events, yet remain amongst the most difficult HABs to effectively monitor for human and environmental health. Monitoring of Karenia blooms is necessary for a variety of precautionary, management and predictive purposes. These include the protection of public health from exposure to aerosolized brevetoxins and the consumption of toxic shellfish, the protection and management of environmental resources, the prevention of bloom associated economic losses, and the evaluation of long term ecosystem trends and for potential future bloom forecasting and prediction purposes. The multipurpose nature of Karenia monitoring, the large areas over which blooms occur, the large range of Karenia cell concentrations (from 5 × 10³ cells L^?1 to >1 × 10⁶ cells L^?1) over which multiple bloom impacts are possible, and limitations in resources and knowledge of bloom ecology have complicated K. brevis monitoring, mitigation and management strategies. Historically, K. brevis blooms were informally and intermittently monitored on an event response basis in Florida, usually in the later bloom stages after impacts (e.g. fish kills, marine mammal mortalities, respiratory irritation) were noted and when resources were available. Monitoring of different K. brevis bloom stages remains the most practical method for predicting human health impacts and is currently accomplished by the state of Florida via direct microscopic counts of water samples from a state coordinated volunteer HAB monitoring program. K. brevis cell concentrations are mapped weekly and disseminated to stakeholders via e-mail, web and toll-free phone numbers and provided to Florida Department of Agriculture and Consumer Services (FDACS) for management of both recreational and commercial shellfish beds in Florida and to the National Oceanic and Atmospheric Administration (NOAA) for validation of the NOAA Gulf of Mexico HAB bulletin for provision to environmental managers. Many challenges remain for effective monitoring and management of Karenia blooms, however, including incorporating impact specific monitoring for the diverse array of potential human and environmental impacts associated with blooms, timely detection of offshore bloom initiation, sampling of the large geographic extent of blooms which often covers multiple state boundaries, and the involvement of multiple Karenia species other than K. brevis (several of which have yet to be isolated and described) with unknown toxin profiles. The implementation and integration of a diverse array of optical, molecular and hybrid Karenia detection technologies currently under development into appropriate regulatory and non-regulatory monitoring formats represents a further unique challenge.     

Exogenous sodium sulfide improves morphological and physiological responses of a hybrid Populus species to nitrogen dioxide

Gaseous nitrogen dioxide (NO₂) can disturb normal plant growth and trigger complex physiological responses. NO₂-induced responses are influenced by biotic or abiotic factors. In this study, we investigated the effects of exogenous sodium sulfide (Na₂S, 5 mmol L⁻¹) on epidermis and stomata related physico-chemical responses of hybrid poplar cuttings (Pouplus alba × P. berolinensis) to gaseous NO₂ (4 μl 1⁻¹) for three time periods (0, 14 and 48 h). We also investigated hydrogen sulfide (H₂S), nitrate-nitrogen and nitrate reductase activity (NR) in control and Na₂S treated plants. Our results showed that NO₂ exposure for 48 h led to the decline of NR, maximal PSII quantum yield (F_v/F_m), net photosynthetic rate (P_n), and dark respiration rate (R_d). The maximum rate for the post-illumination carbon dioxide burst (PIB) occurred in 48-h exposed leaves 13–15 s after darkening. Moreover, NO₂ exposure resulted in a significant increase in nitrogen percentage (from 0 to 33%) and a decrease in the macro and micro-elements of leaf surface. Spraying Na₂S aqueous solution on the leaf surfaces significantly increased the thicknesses of palisade/spongy tissue and H₂S content. Na₂S pretreatment alleviated NO₂-caused toxic effects as indicated by increased NR and higher values of P_n, F_v/F_m, and actual photochemical efficiency in light (ФPSII) compared with the control. Na₂S pretreatment had no significant impacts on PIB-based photorespiration or elements composition of a leaf surface.     

Pea plant responsiveness under elevated [CO2] is conditioned by the N source (N2 fixation versus NO3− fertilization)

The main goal of this study was to test the effect of [CO₂] on C and N management in different plant organs (shoots, roots and nodules) and its implication in the responsiveness of exclusively N₂-fixing and NO₃⁻-fed plants. For this purpose, exclusively N₂-fixing and NO₃⁻-fed (10 mM) pea (Pisum sativum L.) plants were exposed to elevated [CO₂] (1000 μmol mol⁻¹ versus 360 μmol mol⁻¹ CO₂). Gas exchange analyses, together with carbohydrate, nitrogen, total soluble proteins and amino acids were determined in leaves, roots and nodules. The data obtained revealed that although exposure to elevated [CO₂] increased total dry mass (DM) in both N treatments, photosynthetic activity was down-regulated in NO₃⁻-fed plants, whereas N₂-fixing plants were capable of maintaining enhanced photosynthetic rates under elevated [CO₂]. In the case of N₂-fixing plants, the enhanced C sink strength of nodules enabled the avoidance of harmful leaf carbohydrate build up. On the other hand, in NO₃⁻-fed plants, elevated [CO₂] caused a large increase in sucrose and starch. The increase in root DM did not contribute to stimulation of C sinks in these plants. Although N₂ fixation matched plant N requirements with the consequent increase in photosynthetic rates, in NO₃⁻-fed plants, exposure to elevated [CO₂] negatively affected N assimilation with the consequent photosynthetic down-regulation.     

Nitrogen uptake and regeneration (ammonium regeneration,nitrification and photoproduction) in waters of the West Florida Shelf prone to blooms of Karenia brevis

The West Florida Shelf (WFS) encompasses a range of environments from inshore estuarine to offshore oligotrophic waters, which are frequently the site of large and persistent blooms of the toxic dinoflagellate, Karenia brevis. The goals of this study were to characterize the nitrogen (N) nutrition of plankton across the range of environmental conditions on the WFS, to quantify the percentage of the plankton N demand met through in situ N regeneration, and to determine whether planktonic N nutrition changes when high concentrations of Karenia are present. In the fall of 2007, 2008, and 2009 we measured ambient nutrient concentrations and used stable isotope techniques to measure rates of primary production and uptake rates of inorganic N (ammonium, NH₄⁺, and nitrate, NO₃⁻), and organic N and carbon (C; urea and amino acids, AA) in estuarine, coastal, and offshore waters, as well as coastal sites with Karenia blooms present. In parallel, we also measured rates of in situ N regeneration – NH₄⁺ regeneration, nitrification, and photoproduction of NH₄⁺, nitrite and AA. Based on microscope observations, ancillary measurements, and previous monitoring history, Karenia blooms sampled represented three bloom stages – initiation in 2008, maintenance in 2007, and late maintenance/stationary phase in 2009. Nutrient concentrations were highest at estuarine sampling sites and lowest at offshore sites. Uptake of NH₄⁺ and NO₃⁻ provided the largest contribution to N nutrition at all sites. At the non-Karenia sites, in situ rates of NH₄⁺ regeneration and nitrification were generally sufficient to supply these substrates equal to the rates at which they were taken up. At Karenia sites, NO₃⁻ was the most important N substrate during the initiation phase, while NH₄⁺ was the most important N form used during bloom maintenance and stationary phases. Rates of NH₄⁺ regeneration were high but insufficient (85 ± 36% of uptake) to support the measured NH₄⁺ uptake at all the Karenia sites although nitrification rates far exceeded uptake rates of NO₃⁻. Taken together our results support the “no smoking gun” nutrient hypothesis that there is no single nutrient source or strategy that can explain Karenia's frequent dominance in the waters where it occurs. Consistent with other papers in this volume, our results indicate that Karenia can utilize an array of inorganic and organic N forms from a number of N sources.     

In situ nutrient monitoring: A tool for capturing nutrient variability and the antecedent conditions that support algal blooms

While increased nutrient concentrations and eutrophication are recognized to be among the causative factors contributing to algal blooms, including harmful algal blooms (HABs), relationships between nutrient fluxes and specific blooms are often not well defined or understood. In an attempt to better decipher these relationships, we employed in situ nutrient monitors to collect time-series data to document variability ranging from hourly to monthly, including rain events and the resulting response of phytoplankton biomass. Multiple deployments are reported here, all conducted in tributaries of Chesapeake Bay. The suite of nutrients that were monitored varied with deployment; examples given here include nitrate + nitrite (NO₃⁻ + NO₂⁻), ammonium (NH₄⁺), phosphate (PO₄³⁻), and urea. Common features in the data included highly varying concentrations on time-scales of a few hours related to tidal oscillations, and longer-term responses on the scale of days related to rainfall events. Increases following rainfall events for all nutrients generally tended to be many fold, up to an order of magnitude, higher than pre-rainfall concentrations. However, the time scale of response to rainfall for individual nutrients varied. Ephemeral increases in PO₄³⁻ and urea typically were contemporaneous with rain events, and were also followed by longer-term sustained increases relative to pre-rainfall levels. Increases in NO₃⁻ + NO₂⁻ and NH₄⁺ lagged rainfall events by a period of several days and generally lasted for several days. These dynamics generally would be missed by traditional manual sampling. Algal responses tended to follow the increases in nitrogen, underscoring nitrogen limitation in these systems even when ambient concentrations were not depleted.     

Alteration of plankton communities and biogeochemical cycles by harmful Cochlodinium polykrikoides (Dinophyceae) blooms

Cochlodinium polykrikoides is a globally distributed, ichthyotoxic, bloom-forming dinoflagellate. Blooms of C. polykrikoides manifest themselves as large (many km²) and distinct patches with cell densities exceeding 10³ ml⁻¹ while water adjacent to these patches can have low cell densities (<100 cells ml⁻¹). While the effect of these blooms on fish and shellfish is well-known, their impacts on microbial communities and biogeochemical cycles are poorly understood. Here, we investigated plankton communities and the cycling of carbon, nitrogen, and B-vitamins within blooms of C. polykrikoides and compared them to areas in close proximity (<100 m) with low C. polykrikoides densities. Within blooms, C. polykrikoides represented more than 90% of microplankton (>20 μm) cells, and there were significantly more heterotrophic bacteria and picoeukaryotic phytoplankton but fewer Synechococcus. Terminal restriction fragment length polymorphism analysis of 16S and 18S rRNA genes revealed significant differences in community composition between bloom and non-bloom samples. Inside the bloom patches, concentrations of vitamin B₁₂ were significantly lower while concentrations of dissolved oxygen were significantly higher. Carbon fixation and nitrogen uptake rates were up to ten times higher within C. polykrikoides bloom patches. Ammonium was a more important source of nitrogen, relative to nitrate and urea, for microplankton within bloom patches compared to non-bloom communities. While uptake rates of vitamin B₁ were similar in bloom and non-bloom samples, vitamin B₁₂ was taken up at rates five-fold higher (>100 pmol⁻¹ L⁻¹ d⁻¹) in bloom samples, resulting in turn-over times of hours during blooms. This high vitamin demand likely led to the vitamin B₁₂ limitation of C. polykrikoides observed during nutrient amendment experiments conducted with bloom water. Collectively, this study revealed that C. polykrikoides blooms fundamentally change microbial communities and accelerate the cycling of carbon, some nutrients, and vitamin B₁₂.     

The effects of light intensity on the growth of Japanese Gambierdiscus spp. (Dinophyceae)

Marine toxic dinoflagellates of the genus Gambierdiscus are the causative agents of ciguatera fish poisoning (CFP), a form of seafood poisoning that is widespread in tropical, subtropical and temperate regions worldwide. The distributions of Gambierdiscus australes, Gambierdiscus scabrosus and two phylotypes of Gambierdiscus spp. type 2 and type 3 have been reported for the waters surrounding the main island of Japan. To explore the bloom dynamics and the vertical distribution of these Japanese species and phylotypes of Gambierdiscus, the effects of light intensity on their growth were tested, using a photoirradiation-culture system. The relationship between the observed growth rates and light intensity conditions for the four species/phylotypes were formulated at R > 0.92 (p < 0.01) using regression analysis and photosynthesis-light intensity (P-L) model. Based on this equation, the optimum light intensity (L_max) and the semi-optimum light intensity range (L_s-opt) that resulted in the maximum growth rate (μ_max) and ≥80% μ _max values of the four species/phylotypes, respectively, were as follows: (1) the L_max and L_s-opt of G. australes were 208 μmol photons m⁻² s⁻¹ and 91–422 μmol photons m⁻² s⁻¹, respectively; (2) those of G. scabrosus were 252 and 120–421 μmol photons m⁻² s⁻¹, respectively; (3) those of Gambierdiscus sp. type 2 were 192 and 75–430 μmol photons m⁻² s⁻¹, respectively; and (4) those of Gambierdiscus sp. type 3 were ≥427 and 73–427 μmol photons m⁻² s⁻¹, respectively. All four Gambierdiscus species/phylotypes required approximately 10 μmol photons m⁻² s⁻¹ to maintain growth. The light intensities in coastal waters at a site in Tosa Bay were measured vertically at 1 m intervals once per season. The relationships between the observed light intensity and depth were formulated using Beer’s Law. Based on these equations, the range of the attenuation coefficients at Tosa Bay site was determined to be 0.058–0.119 m⁻¹. The values 1700 μmol photons m⁻² s⁻¹, 500 μmol photons m⁻² s⁻¹, and 200 μmol photons m⁻² s⁻¹ were substituted into the equations to estimate the vertical profiles of light intensity at sunny midday, cloudy midday and rainy midday, respectively. Based on the regression equations coupled with the empirically determined attenuation coefficients for each of the four seasons, the ranges of the projected depths of L_max and L_s-opt for the four Gambierdiscus species/phylotypes under sunny midday conditions, cloudy midday conditions, and rainy midday conditions were 12–38 m and 12–54 m, 1–16 m and 1–33 m, and 0 m and 0–16 m, respectively. These results suggest that light intensity plays an important role in the bloom dynamics and vertical distribution of Gambierdiscus species/phylotypes in Japanese coastal waters.