Physiological changes in the coastal marine cyanobacterium Synechococcus sp. PCC 7002 exposed to low ferric ion levels

Cyanobacteria are ubiquitous in marine waters. These prokaryotic cells are of particular interest in areas of the ocean where the availability of iron may be limiting for cell growth since these organisms commonly excrete iron-specific organic ligands (siderophores) in response to low levels of iron. It is generally considered that the production of siderophores provides a competitive advantage over the competing microorganisms that do not produce these ligands.In order to ascertain the influence of iron availability on the physiology of picoplanktonic cyanobacteria we performed a series of experiments on the coastal coccoid cyanobacterium, Synechococcus sp. PCC 7002. Physiological responses were examined in cells grown in a continuous cuture system with influx media containing a range of iron concentrations (from 4.2 × 10⁻⁵ to 5.1 × 10⁻⁹ M FeCl₃). Steady-state growth rates, combined with growth data from batch cultures demonstrated a non-linear response between iron availability and cell proliferation: cell yields were considerably higher in the lowest-iron chemostats than predicted based on the yields in the higher-iron chemostats. The higher yields during low-iron growth corresponded with the production of the extracellular siderophores and the induction of the specific iron-siderophore membrane transport proteins. A comparison of iron transport and carbon acquisition rates between the low-iron grown cells and the high-iron grown cells indicates that under low-iron growth conditions, iron and carbon acquisition meets the growth demands of the cells, whereas growth at higher iron levels enabled excessive (luxury) carbon acquisition and storage. We conclude that cyanobacteria are efficiently adapted to grow in low-iron environments (providing sufficient light for photosynthesis is available) and the luxury-uptake of carbon may serve as the source material for the extracellular ligands released by these cells. Since the release of siderophores was at iron levels in excess of the levels that induce the siderophore-mediated transport of iron, cyanobacteria growing in an environment with varying levels of iron may contribute substantial amounts of their stored carbon reserves into the DOC as iron-specific ligands.     

The exceptionally stable cobalt(III)–desferrioxamine B complex

The biogeochemistry of trivalent iron, manganese, and cobalt in the oceans is dominated by soluble complexes formed with high-affinity organic ligands that are believed to be microbial siderophores or similar biogenic chelating agents. Desferrioxamine B (DFOB), a trihydroxamate siderophore found in both terrestrial and marine environments, has served as a useful model for a large class of microbial siderophores in studies of 1:1 complexes formed with trivalent iron and manganese. However, no data exist concerning DFOB complexes with Co(III), which we hypothesize should be as strong as those with Fe(III) and Mn(III) if the current picture of the ocean biogeochemistry of the three trivalent metals is accurate. We investigated the complexation reaction between DFOB and Co(III) in aqueous solution at seawater pH using base and redox titrations, and then characterized the resulting 1:1 complex Co(III)HDFOB⁺ using X-ray absorption, resonance Raman spectroscopy, and quantum mechanical structural optimizations. We found that the complex stability constant for Co(III)HDFOB⁺ (log K _{[Co(III)HDFOB⁺]} = 37.5 ± 0.4) is in fact five and seven orders of magnitude larger than that for Fe(III)HDFOB⁺ (log K_{[Fe(III)HDFOB⁺]} = 32.02) and Mn(III)HDFOB⁺ (log K_{[Mn(III)HDFOB}⁺_] = 29.9), respectively. Spectroscopic data and the supporting theoretical structural optimizations elucidated the molecular basis for this exceptional stability. Although not definitive, our results nevertheless are consistent with the evolution of siderophores as a response by bacteria to oxygenation, not only because of sharply decreasing concentrations of Fe(III), but also of Co(III).     

On the relevance of iron adsorption to container materials in small-volume experiments on iron marine chemistry: Fe-aided assessment of capacity, affinity and kinetics

Iron chemistry in seawater has been extensively studied in the laboratory, mostly in small-volume sample bottles. However, little has been reported about iron wall sorption in these bottles. In this paper, radio-iron ⁵⁵Fe was used to assess iron wall adsorption, both in terms of capacity, affinity and kinetics. Various bottle materials were tested. Iron sorption increased from polyethylene/polycarbonate to polymethylmetacrylate (PMMA)/high-density polyethylene/polytetrafluoroethylene to glass/quartz, reaching equilibrium in a 25–70 h period. PMMA was studied in more detail: ferric iron (Fe(III)) adsorbed on the walls of the bottles, whereas ferrous iron (Fe(II)) did not. Considering that in seawater the inorganic iron pool mostly consists of ferric iron, the wall will be a factor that needs to be considered in bottle experiments.The present data indicate that for PMMA with specific surface (S)-to-volume (V) ratio S/V, both iron capacity (42 ± 16 × 10^− 9 mol/m² or 1.7 × 10^− 9 mol/L recalculated for the S/V-specific PMMA bottles used) and affinity (log K_Fe'W = 11.0 ± 0.3 m²/mol or 12.4 ± 0.3 L/mol, recalculated for the S/V-specific PMMA bottles used) are of similar magnitude as the iron capacity and -affinity of the natural ligands in the presently used seawater and thus cannot be ignored.Calculation of rate constants for association and dissociation of both Fe'L (iron bound to natural occurring organic ligands) and Fe'W (iron adsorbed on the wall of vessels) suggests that the two iron complexes are also of rather similar kinetics, with rate constants for dissociation in the order of 10 ⁻⁴–10^− 5 L/s and rate constants for association in the order of 10⁸ L/(mol s). This makes that iron wall sorption should be seriously considered in small-volume experiments, both in assessments of shorter-term dynamics and in end-point observations in equilibrium conditions. Therefore, the present data strongly advocate making use of iron mass balances throughout in experiments in smaller volume set-ups on marine iron (bio) chemistry.     

A comparison of two voltammetric techniques for determining zinc speciation in Northeast Pacific Ocean waters

Two independent voltammetric techniques, differential pulse cathodic stripping voltammetry (DPCSV) and differential pulse anodic stripping voltammetry (DPASV), determined that 95% of the dissolved zinc is organically complexed at two depths (60 and 150 m) within the surface euphotic zone at an open ocean station in the Northeast Pacific. Average values for the concentrations of the natural zinc-complexing organic ligands (C_L) obtained from duplicate determinations at these two depths by DPCSV versus DPASV are in excellent agreement: 1.60 ± 0.01 versus 1.76 ± 0.03 nM at 60 m, and 2.14 (n=1) versus 2.22 ± 0.06 nM at 150 m. Average values for the conditional stability constants (with respect to free Zn²⁺) of the natural zinc-organic complexes (log K^′_ZnL) from duplicate determinations at both depths by DPCSV versus DPASV are 10.3 ± 0.2 versus 11.2 ± 0.2. Additional research is required to assess the significance of the difference in the conditional stability constants determined by these two techniques. These results confirm recent observations that strong zinc complexes formed with an organic ligand class existing at nanomolar concentrations dominates zinc speciation in the North Pacific.     

Investigations of iron coordination and redox reactions in seawater using Fe radiometry and ion-pair solvent extraction of amphiphilic iron complexes

Iron coordination and redox reactions in synthetic and coastal seawater were investigated at nanomolar concentrations using ⁵⁹Fe radiometry and ion-pair solvent extraction of iron chelated by sulfoxine (8-hydroxyquinoline-5-sulfonate) and BPDS (bathophenanthroline disulfonate). Using sulfoxine, we determined the rate at which the monomeric Fe(III) hydroxide species present in seawater of pH 8 are complexed by the microbial siderophore deferriferrioxamine B and the synthetic chelator EDTA (ethylenediaminetetraacetic acid). Forward rate constants of 2 × 10⁶M⁻¹s⁻¹ and 20 M⁻¹s⁻¹, respectively, were obtained. The kinetics of these reactions have not been measured previously at pH values near that of seawater. Conditional equilibrium constants measured for the Fe(III)-EDTA system are consistent with published stability constants for EDTA complexes and for Fe(III) hydrolytic equilibria minus the neutral Fe(OH)₃^o species, suggesting it is not quantitatively significant near pH 8. Commercial humic acid was found to have sufficient affinity for iron to compete with Fe(III) hydrolysis in seawater, and limited evidence was obtained for an interaction with dissolved organic matter in coastal seawater.In our investigations of redox reactions using BPDS to trap Fe(II) produced in the medium, we observed enhanced photoreduction of Fe(III) by humic acid as well as reduction induced by solutes released from phytoplankton in seawater of pH 8. Although the method is sensitive enough to work at near-oceanic levels of iron, the difficulty in distinguishing Fe(II) generated by Fe(III)-BPDS interactions from Fe(II) produced by other means limits its utility. This analytical ambiguity may be generalizable to other methods which measure ferrous iron in seawater using Fe(II)-specific ligands.     

Effect of siderophores on the light-induced dissolution of colloidal iron(III) (hydr)oxides

Siderophores play an important role in biological iron acquisition in iron-limited aquatic systems. While it is widely accepted that the solubilization of iron-bearing mineral phases is a key function of siderophores, the mechanism of siderophore-promoted mineral dissolution in aquatic systems is largely unknown. In this study, we investigated the effect of siderophores (desferrioxamine B (DFOB) and aerobactin) on light-induced dissolution of goethite and lepidocrocite in the presence or absence of oxalate in aerated and deaerated suspensions at pH 6. For the irradiated two-ligand system (oxalate/siderophore), the experimental results suggest that oxalate acts as the electron donor for the formation of surface Fe(II), and the siderophore acts as an efficient shuttle for the transfer of surface Fe(II) into solution. Furthermore, even in the absence of an electron donor such as oxalate, both DFOB and aerobactin accelerated the light-induced dissolution of lepidocrocite as compared to the thermal dissolution. Experiments with dissolved Fe(III)–DFOB and Fe(III)–aerobactin complexes suggest that this enhancing effect is not due to photolysis of corresponding surface complexes but to efficient transfer of reduced surface Fe(II) into solution, where surface Fe(II) may be formed, e.g., through photolysis of surface Fe(III)–hydroxo groups. Based on this study, we conclude that the interplay of light and siderophores may play a key role in the dissolution of colloidal iron(III) (hydr)oxides in marine systems, particularly in the presence of efficient electron donors.     

Characterization of the binding sites for Al(III) and Be(II) in a sample of marine fulvic acids

A sample of coastal marine fulvic acids (mfua) was studied and its binding site structures that form stable complexes with metal ions were identified. A previously developed self-modeling methodology, based on the coupling of synchronous fluorescence (SyF) spectroscopy with evolving factor analysis (EFA), was used for the analysis of the interactions between mfua and two metal ions that act as probes for binding sites, namely Be(II) and Al(III). Two types of binding site structures were detected, probably of the salicylic acid and catechol types. The values of the conditional stability constants between mfua and the two metal ions were: for Be(II), at pH = 6, log K = 5.32(8); for Al(III), at pH = 4, log K = 5.1(2). The concentration of the corresponding binding sites was found to be less than one half of those found for soil fulvic acid samples.     

Organic complexation and its control of the dissolved concentrations of copper and zinc in the Scheldt estuary

Cathodic stripping voltammetry (CSV) is used to determine total (after UV-irradiation) and labile dissolved metal concentrations as well as complexing ligand concentrations in samples from the river Scheldt estuary. It was found that even at high added concentrations of catechol (1 m for copper and 0·4 m for iron) and of APDC (1 m for zinc) only part of the dissolved metal was labile (5–58% for copper, 34–69% for zinc, 10–38% for iron); this discrepancy could be explained by the low solubility of iron which is largely present as colloidal material, and by competition for dissolved copper and zinc by organic complexing ligands. Ligand concentrations varied between 28 and 206 n for copper and between 22 and 220 n for zinc; part of the copper complexing ligands could be sub-divided into strong complexing sites with concentrations between 23 and 121 n and weaker sites with concentrations between 44 and 131 n . Values for conditional stability constants varied between (logK′ values) 13·0 and 14·8 for strong and between 11·5 and 12·1 for weaker copper complexing ligands, whereas for zinc the values were between 8·6 and 10·6. The average products of ligand concentrations and conditional stability constants (a-coefficients) were 6 × 10² for zinc and 6 × 10⁶ for copper.The dissolved zinc concentration was found to co-vary with the zinc complexing ligand concentration throughout the estuary. It is argued that the zinc concentration is regulated, in this estuary at least, by interactions with dissolved organic complexing ligands. A similar relationship was apparent between the dissolved copper and the strong copper complexing ligand concentration. The total copper complexing ligand concentrations were much greater than the dissolved copper concentrations, suggesting that only strongly complexed copper is kept in solution.These results provide evidence for the first time that interactions of copper and zinc with dissolved organic complexing ligands determine the geochemical pathway of these metals.     

Determination of conditional stability constants of organic copper and zinc complexes dissolved in seawater using ligand exchange method with EDTA

To clarify the nature of organic metal complexes dissolved in seawater, a ligand exchange reaction between ligands of natural origin and an aminopolycarboxylic acid (EDTA) was used to determine the conditional stability constants of organic metal complexes. The results indicate that more than two organic molecules complexed with copper and zinc exist in surface seawater. It is found that the conditional stability constants of these naturally-occurring organic metal complexes are 1–3 orders of magnitude higher than those of EDTA-Cu and EDTA-Zn complexes. These estimates of the conditional stability constants for the dominant species of organic copper and zinc complexes are 10^11.8 and 10^9.3, respectively, at pH 8.1. The results indicate that these naturally-occurring organic metal complexes are stable species and not easily dissociated or displaced with others in the marine environment.     

The speciation of Fe(II) and Fe(III) in natural waters

The interactions of Fe(II) and Fe(III) with the inorganic anions of natural waters have been examined using the specific interaction and ion pairing models. The specific interaction model as formulated by Pitzer is used to examine the interactions of the major components (Na⁺, Mg²⁺, Ca²⁺, K⁺, Sr²⁺, Cl⁻, SO₄⁻, HCO₃⁻, Br⁻, CO₃²⁻, B(OH)₄⁻, B(OH)₃ and CO₂) of seawater and the ion pairing model is used to account for the strong interaction of Fe(II) and Fe(III) with major and minor ligands (Cl⁻, SO₄²⁻, OH⁻, HCO₃⁻, CO₃²⁻ and HS⁻) in the waters. The model can be used to estimate the activity and speciation of iron in natural waters as a function of composition (major sea salts) and ionic strength (0 to 3 M). The measured stability constants (K_FeX^*) of Fe(II) and Fe(III) have been used to estimate the thermodynamic constants (K_FeX) and the activity coefficient of iron complexes (γ_FeX) with a number of inorganic ligands in NaClO₄ medium at various ionic strengths: In(K_FeX/γ_Feγ_X) = InK_FeX − In(γ_FeX) The activity coefficients for free ions (γ_Fe, γ_x) needed for this extrapolation have been estimated from the Pitzer equations. The activity coefficients of the ion pairs have been used to determine Pitzer parameters (B_FeX, B_FeX⁰, C_FeX^φ) for the iron complexes. These results make it possible to estimate the stability constants for the formation of Fe(II) and Fe(III) complexes over a wide range of ionic strengths and in different media. The model has been used to determine the solubility of Fe(III) in seawater as a function of pH. The results are in good agreement with the measurements of Byrne and Kester and Kuma et al. When the formation of Fe organic complexes is considered, the solubility of Fe(III) in seawater is increased by about 25%.     

The reliability of the thermodynamic constants for the dissociation of carbonic acid in seawater

Laboratory measurements of all four CO₂ parameters [f_CO2 ( = fugacity of CO₂), pH, TCO₂ ( = total dissolved inorganic carbon), and TA ( = total alkalinity)] were made on the same sample of Gulf Stream seawater (S = 35) as a function of temperature (5–35 °C) and the ratio of TA/TCO₂ (X) (1.0–1.2). Overall the measurements were consistent to ±8 μ atm in f_CO2, ± 0.004 in pH, ± 3 μ mol kg⁻¹ in TCO₂, and ± 3 μ mol kg⁻¹ in TA with the thermodynamic constants of Goyet and Poisson (1989), Roy et al. (1993), and Millero (1995). Deviations between the measured pH, TCO₂, TA and those calculated from various input combinations increase with increasing X when the same constants are used. This trend in the deviations indicates that the uncertainties in pK₂ become important with increasing X (surface waters), but are negligible for samples with the lower X (deep waters). This trend is < 5 μ mol kg⁻¹ when the pK₂ values of Lee and Millero (1995) are used.The overall probable error of the calculated f_CO2 due to uncertainties in the accuracy of the parameters (pH, TCO₂, TA, pK₀, pk₁, and pK₂) is ± 1.2%, which is similar to the differences between the measured values and those calculated using the thermodynamic constants of Millero (1995).The calculated values of pK₁, (from f_CO2-TCO₂-TA) agree to within ± 0.004 compared to the results of Dickson and Millero (1987), Goyet and Poisson (1989), Roy et al. (1993), and Millero (1995) over the same experimental conditions. The calculated values of pK₂ (from pH-TCO₂-TA) are in good agreement (± 0.004) with the results of Lee and Millero (1995) and also in reasonable agreement (± 0.008) with the results of Goyet and Poisson (1989), Roy et al. (1993), and Millero (1995). The salinity dependence of our derived values of pK₁ and pK₂, (S = 35) can be estimated using the equations determined by Millero (1995).     

Glutathione S-transferase from an antarctic fish, Dissostichus mawsoni

Glutathione S-transferase activity was measured in the hepatic cytosol front Dissostichus mawsoni and Pagothenia borchgrevinki. Activity measures with 1-chloro-2,4-dinitrobenzene as substrate were 11·2 and 16·7 μmol/min/g tissue respectively. Little or no activity was detected when p-nitrobenzyl chloride or 3,4-dichloro-1-nitrobenzene were used as substrate. The hepatic glutathione S-transferases from D. mawsoni were partially purified using gel filtration and chromatofocusing. Three peaks of activity were resolved. The major isozyme (158-fold purification) eluting at pH7·1 appeared to be catalytically a homodimer. The isozyme was highly inhibited by triphenyltin chloride (IC₅₀ = 0·1 μ) while inhibition constants for Cibicron Blue 3GA, bromosulphophalein and hematin were 1·1, 20 and 34 μ respectively.     

Meridional distribution and carbon biomass of autotrophic picoplankton in the Central North Pacific Ocean during late northern summer 1990

The meridional distribution of autotrophic picoplankton groups in the central north Pacific was studied during the late northern summer of 1990. Sampling was along a section at 175°N which extended from 45°N to 8°S. The section is far from coastal regions and included subarctic, central gyre, and equatorial areas. Five autotrophic picoplankton groups, autotrophic microflagellate, red-fluorescing picoplankton,Synechococcus, prochlorophyte, and orange-fluorescing picoplankton, were identified from samples taken at stations distributed along this section. These five groups showed distinctive differences in their meridional and vertical distributions. The autotrophic microflagellates and red-fluorescing picoplankton showed distributions that were similar to that of chlorophyll a, which was dominated by the <3 μm size fraction. However, the vertical distribution of these groups was different.Synechococcus was found mostly in surface waters (PAR<10%) and was particularly abundant in the Kuroshio Extension and south of the equatorial region where the nitracline was shallow (50–75 m). Prochlorophytes were abundant in the deep euphotic layer (PAR 1-0.1%) from the south of the Kuroshio Extension to the south of the equatorial area. Orange-fluorescing picoplankton, which may be one kind of cyanobacteria but is larger than typical Synechococcus, were mostly distributed in the oligotrophic surface waters of the central gyre. The carbon biomass estimates for these organisms showed that these five groups dominated in different areas. The vertical distribution of carbon biomass did not correspond to that of chlorophyll a in the central gyre and south of the equator because of the larger carbon/ chlorophyll a ratio of Synechococcus and orange-fluorescing picoplankton relative to that of the other picoplankton.     

Long-term effect of beach replenishment on natural recovery of shallow Posidonia oceanica meadows

The recovery capacity of shallow Posidonia oceanica meadows degraded by beach replenishment eighteen years before was assessed in two impacted meadows and compared with other two undisturbed localities. Inside each locality, we selected randomly three sites separated by 500–1000 m. At site level we study the vitality of P. oceanica meadow assessing the vegetative growth, leaf characteristics, and non-structural carbohydrates of the plants. Additionally, at locality level, silt-clay fraction, organic matter, pH and light intensity incident on the sea bottom were measured to evaluate the environmental conditions. Covering of P. oceanica was significantly lower at the impacted localities while amount of dead “matte” was higher. Leaf production of horizontal rhizomes (14.6 ± 1.11 vs 19.47 ± 1.45 leaves y⁻¹), net total rhizomes recruitment (2.33 ± 0.17 vs 4.3 ± 0.33 branches y⁻¹) and starch concentration (43.625 ± 0.67 vs 54.45 ± 0.74 mg per g of rhizome) at impacted meadows were significantly lower than controls. Leaf features, epiphytes biomass, colonization, elongation and horizontal and vertical rhizome production did not show significant differences. Sediments at impacted localities contained higher silt-clay fraction and higher organic matter load while pH was lower. Light intensity on the sea bottom measured at all localities was over the minimum light requirements estimated for P. oceanica. Our results show that the press impact produced by beach replenishment was enduring in the time slowing natural recovery by 45%. This impact may be related with changes in the sediment features.     

Transformation dynamics and reactivity of dissolved and colloidal iron in coastal waters

We have investigated the chemical forms, reactivities and transformation kinetics of Fe(III) species present in coastal water with ion exchange and filtration methods. To simulate coastal water system, a mixture of ferric iron and fulvic acid was added to filtered seawater and incubated for a minute to a week. At each incubation time, the seawater sample was acidified with hydrochloric acid and then applied to anion exchange resin (AER) to separate negatively charged species (such as fulvic acid, its complexes with iron and iron oxyhydroxide coated with fulvic acid) from positively charged inorganic ferric iron (Fe(III)′). By monitoring the acid-induced Fe(III)′ over an hour, it was found that iron complexed by fulvic acid dissociated rapidly to a large extent (86–92% at pH 2), whereas amorphous ferric oxyhydroxide particles associated with fulvic acid (AFO-L) dissociated very slowly with the first-order dissociation rate constants ranging from 6.1 × 10^− 5 for pH 3 to 2.7 × 10^− 4 s^− 1 for pH 2. Therefore, a brief acidification followed by the AER treatment (acidification/AER method) was likely to be able to determine fulvic acid complexes and thus differentiate the complexes from the AFO-L particles (the dissolution of AFO-L was insignificant during the brief acidification). The acidification/AER method coupled with a simple filtration technique suggested that the iron–fulvic acid complexes exist in both the < 0.02 μm and 0.02–0.45 μm size fractions in our coastal water system. The truly dissolved iron (< 0.02 μm) was relatively long-lived with a life-time of 14 days, probably due to the complexation by strong ligands. Such an acid-labile iron may be an important source of bioavailable iron in coastal environments, as a significant relationship between the chemical lability and bioavailability of iron has been well recognised.     

Spectrophotometric pH measurement in estuaries using thymol blue and m-cresol purple

The conditional acid dissociation constants (pK_a′) of two sulfonephthalein dyes, thymol blue (TB) and m-cresol purple (mCP), were assessed throughout the estuarine salinity range (0<S<40) using a tris/tris–HCl buffer and spectrophotometric measurement. The salinity dependence of the pK_a′ of both dyes was fitted to the equations (25 °C, total proton pH scale, mol kg soln⁻¹):

The estimated accuracy of pH measurements using these calculated pK_a′ values is considered to be comparable to that possible with careful use of a glass electrode (±0.01 pH unit) but spectrophotometric measurements in an estuary have the significant advantage that it is not necessary to calibrate an electrode at different salinities. pH was measured in an estuary over a tidal cycle with a precision of ±0.0005 pH unit at high (S>30) salinity, and ±0.002 pH unit at low (S<5) salinity. The pH increased rapidly in the lower salinity ranges (0<S<15) but less rapidly at higher salinities.     

Determination of Henry's law constants for hexachlorocyclohexanes in distilled water and artificial seawater as a function of temperature

Henry's law constants were determined for α- and γ-hexachlorocyclohexane (HCH) as a function of temperature (0.5–45°C) in artificial seawater (SW; 30‰) and distilled water (DW) using the gas stripping method. Water samples (1–5 ml) were withdrawn from the stripping vessel during the stripping process (30–360 h), solvent extracted and analyzed by gas chromatography—electron-capture detection. The effect of bubbling depth was checked to ensure that bubbles leaving the system were at equilibrium with HCHs in the aqueous phase. Henry's law constants determined at 35 and 45°C in SW were significantly higher (P≤ 0.05) than in DW for both α- and γ-HCH, but not at lower temperatures. The slopes (m) and intercepts (b) of log H vs. 1 / T plots were: α-HCH (DW, 0.5–45°C); m = −2810 ± 110, B = 9.31 ± 0.38; α-HCH (SW, 0.5–23°C); M = −2969 ± 218, B = 9.88 ± 0.76; γ-HCH (DW, 0.5–45°C); M = −2382 ± 160, B = 7.54 ± 0.54; γ-HCH (SW, 0.5–23°C); M = −2703 ± 276, B = 8.68 ± 0.96. Henry's law constants determined in this study compared well with those calculated from reported vapor pressure and solubility data.     

Impact of environmental factors on in situ determination of iron in seawater by flow injection analysis

A sensitive method for iron determination in seawater has been adapted on a submersible chemical analyser for in situ measurements. The technique is based on flow injection analysis (FIA) coupled with spectrophotometric detection. When direct injection of seawater was used, the detection limit was 1.6 nM, and the precision 7%, for a triplicate injection of a 4 nM standard. At low iron concentrations, on line preconcentration using a column filled with 8-hydroxyquinoline (8HQ) resin was used. The detection limit was 0.15 nM (time of preconcentration = 240 s), and the precision 6%, for a triplicate determination of a 1 nM standard, allowing the determination of Fe in most of the oceanic regimes, except the most depleted surface waters. The effect of temperature, pressure, salinity, copper, manganese, and iron speciation on the response of the analyser was investigated. The slope of the calibration curves followed a linear relation as a function of pressure (C_p = 2.8 × 10^{− 5}P + 3.4 × 10^{− 2} s nmol^{− 1}, R² = 0.997, for Θ = 13 °C) and an exponential relation as a function of temperature (C_Θ = 0.009e^0.103Θ, R² = 0.832, for P = 3 bar). No statistical difference at 95% confidence level was observed for samples of different salinities (S = 0, 20, 35). Only very high concentration of copper (1000 × [Fe]) produced a detectable interference. The chemical analyser was deployed in the coastal environment of the Bay of Brest to investigate the effect of iron speciation on the response of the analyser. Direct injection was used and seawater samples were acidified on line for 80 s. Dissolved iron (DFe, filtered seawater (0.4 μm), acidified and stored at pH 1.8) corresponded to 29 ± 4% of Fe_a (unfiltered seawater, acidified in line at pH 1.8 for 80 s). Most of Fe_a (71 ± 4%) was probably a fraction of total dissolvable iron (TDFe, unfiltered seawater, acidified and stored at pH 1.8).     

Association of picophytoplankton distribution with ENSO events in the equatorial Pacific between 145°E and 160°W

Climatological variability of picophytoplankton populations that consisted of >64% of total chlorophyll a concentrations was investigated in the equatorial Pacific. Flow cytometric analysis was conducted along the equator between 145°E and 160°W during three cruises in November–December 1999, January 2001, and January–February 2002. Those cruises were covering the La Niña (1999, 2001) and the pre-El Niño (2002) periods. According to the sea surface temperature (SST) and nitrate concentrations in the surface water, three regions were distinguished spatially, viz., the warm-water region with >28 °C SST and nitrate depletion (<0.1 μmol kg⁻¹), the upwelling region with <28 °C SST and high nitrate (>4 μmol kg⁻¹) water, and the in-between frontal zone with low nitrate (0.1–4 μmol kg⁻¹). Picophytoplankton identified as the groups of Prochlorococcus, Synechococcus and picoeukaryotes showed a distinct spatial heterogeneity in abundance corresponding to the watermass distribution. Prochlorococcus was most abundant in the warm-water region, especially in the nitrate-depleted water with >150×10³ cells ml⁻¹, Synechococcus in the frontal zone with >15×10³ cells ml⁻¹, and picoeukaryotes in the upwelling region with >8×10³ cells ml⁻¹. The warm-water region extended eastward with eastward shift of the frontal zone and the upwelling region during the pre-El Niño period. On the contrary, these regions distributed westward during the La Niña period. These climatological fluctuations of the watermass significantly influenced the distribution of picophytoplankton populations. The most abundant area of Prochlorococcus and Synechococcus extended eastward and picoeukaryotes developed westward during the pre-El Niño period. The spatial heterogeneity of each picophytoplankton group is discussed here in association with spatial variations in nitrate supply, ambient ammonium concentration, and light field.