An optimized method for automated analysis of algal pigments by HPLC

A recent development in algal pigment analysis by high-performance liquid chromatography (HPLC) is the application of automation. An optimization of a complete sampling and analysis protocol applied specifically in automation has not yet been performed. In this paper we show that automation can only be successful if the various methodological aspects of the sampling and analysis protocol are considered in coherence. We introduce an optimized protocol that involves freeze-drying of the sample, subsequent extraction in 90% acetone and the application of water-packing during analysis. The method was evaluated on both natural plankton populations and a broad spectrum of microalgal cultures: Thalassiosira weisflogii (Bacillariophyceae), Emiliania huxleyi (Prymnesiophyceae), Phaeocystis globosa and Phaeocystis antarctica (Prymnesiophyceae) and Pyramimonas sp. (Prasinophyceae). Whereas pigment extracts were unstable in methanol, with recorded chlorophyll a losses from 10% to 60% per day, pigment degradation rates in acetone were generally less than 1% over 18 h storage in the autosampler (4 °C). In addition, it was found that the extraction efficiency of acetone significantly increased upon freeze-drying prior to extraction. Increases as high as 50–60% were measured in P. antarctica. The application of water-packing of the sample during injection resulted in improved peak shape and peak separation, without diluting the pigment concentrations. Automation is especially beneficial for application in the field, when mixed algal assemblages and low biomass put a high demand on the sensitivity as well as reproducibility of the method.     

Effects of salinity, temperature and food type on the uptake and elimination rates of cd, cr, and zn in the asiatic clamcorbicula fluminea

Laboratory radiotracer experiments were conducted to determine assimilation efficiencies (AE) from ingested algal food and oxic sediment particles, uptake rates from the dissolved phase, and the efflux rates of Cd, Cr and Zn in the Asiatic clamCorbicula fluminea. Among three elements, AE from both algal and sediment food was greatest for Cd, followed by Zn and Cr. The AEs of tested elements from algal food(Phaeodactylum tricornutum) were consistently higher than those from sediments at a given salinity and temperature. The influence of salinity (0, 4 and 8 psu) and temperature (5, 13 and 21¼) on the metal AEs was not evident for most tested elements, except Cd AEs from sediment. The rate constant of metal uptake from the dissolved phase (k _u was greatest for Cd, followed by Zn and Cr in freshwater media. However, in saline water, thek _u of Zn were greater than those of Cd. The influx rate of all tested metals increased with temperature. The efflux rate constant was greatest for Cr (0.02 d^-1), followed by Zn (0.010~0.017 d^-1) and Cd (0.006 d^-1). The efflux rate constant for Zn in clam tissues depurated in 0 psu (0.017 d^-1) was faster than that in 8 psu (0.010 d^-1). Overall results showed that the variation of salinity and temperature in estuarine systems can considerably influence the metal bioaccumulation potential in the estuarine clamC. fluminea. The relatively high Cd accumulation capacity ofC. fluminea characterized by the high AE, high dissolved influx rate and low efflux rate, suggested that this clam species can be used as an efficient biomonitor for the Cd contamination in freshwater and estuarine environments.     

Cloning of cytochrome P450 1A (CYP1A) genes from the hermaphrodite fish Rivulus marmoratus and the Japanese medaka Oryzias latipes

To use two small fish Rivulus marmoratus (Cyprinodontiformes, Rivulidae) and the Japanese medaka Oryzias latipes (Belloniformes) as testing models in molecular ecotoxicology, we have cloned the cytochrome P450 1A (CYP1A) gene after screening of both genomic DNA libraries, and sequenced 11,863 and 7,243 bp including all the exons and introns with promoter regions, respectively. The Rivulus and the medaka CYP1A gene consisted of seven exons (including non-coding exons) with high homology to mammals. In the promoter region, Rivulus CYP1A gene has seven xenobiotic response elements (XREs) and two metal response elements (MREs), while the Japanese medaka CYP1A gene has six XREs and four MREs. Interestingly, medaka CYP1A gene has a number of MREs at the promoter, which may affect its response on metal exposure. We describe here the gene structure of both fish CYP1A genes.     

Application of the Single-Bubble Airgun Technique for OBS-Data Acquisition Across the Jan Mayen Ridge,North Atlantic

A 140 km long wide-angle seismic profile has been acquired by use of 6 Ocean Bottom Seismographs across the Jan Mayen Ridge, North Atlantic. The profile was acquired twice; once with a traditionally tuned standard source and secondly with a somewhat smaller source tuned on the first bubble pulse. Analysis of the frequency content of the data reveals that the single-bubble source within the 10-15 Hz frequency range generates a signal with a level about 5 db above that of the standard source. These differences can partly be related to differences in airgun depth. The higher output level for these frequencies enables the single-bubble source to resolve intra-crustal structures with a higher degree of certainty, when compared to the data acquired by use of the standard source array. The standard source seems to generate slightly more energy for frequencies around 6 Hz, probably due to the use of a large 1200 in/sup3 gun in this array. These low frequencies a re of importance for mapping of lower crustal and upper mantle structures, and it is recommended that this is taken into account when seismic sources for mapping of deep crustal and upper mantle structures are designed.     

Sheet flow and suspension of sand in oscillatory boundary layers

after revisionTime-dependent measurements of flow velocities and sediment concentrations were conducted in a large oscillating water tunnel. The measurements were aimed at the flow and sediment dynamics in and above an oscillatory boundary layer in plane bed and sheet-flow conditions. Two asymmetric waves and one sinusoidal wave were imposed using quartz sand with D₅₀ = 0.21 mm. A new electro-resistance probe with a large resolving power was developed for the measurement of the large sediment concentrations in the sheet-flow layer. The measurements revealed a three layer transport system consisting of a pick-up/deposition layer, an upper sheet flow layer and a suspension layer.In the asymmetric wave cases the total net transport was directed “onshore” and was mainly concentrated in the thin sheet flow layer (< 0.5 cm) at the bed. A small net sediment flux was directed “offhore” in the upper suspension layer. The measured flow velocities, sediment concentrations and sedimenl fluxes showed a good qualitative agreement with the results of a (numerical) 1DV boundary-layer flow and transport model. Although the model did not describe all the observed processes in the sheet-flow and suspension layer, the computational results showed a reasonable agreement with measured net transport rates in a wide range of asymmetric wave conditions.     

A new method for the generation of second-order random waves

In order to prevent the generation of spurious free sub- and superharmonics of random waves in a laboratory channel, the control signal for the wave board has to be derived according to a higher-order wave theory. An expression for this control signal has been derived with the perturbation method of multiple scales. It is much less complex and requires less computation time than the expressions obtained from the full second-order theory. The new method for second-order subharmonics was verified experimentally for waves with bichromatic and continuous first-order spectra. The data were analysed with the complex-harmonic principal-component analysis to reduce the influence of noise.     

Measurement of the redox speciation of iron in seawater by catalytic cathodic stripping voltammetry

Catalytic cathodic stripping voltammetry (CSV) preceded by adsorptive collection of complexes of 1-nitroso-2-napthol (NN) can be used to determine iron in seawater. It is shown here that iron(II) is effectively masked in the presence of 2,2-dipyridyl (Dp) so that iron(III) is measured selectively. The concentration of iron(II) is then calculated as the difference between the concentrations of reactive iron (Fe_R) in the absence and presence of 2 μM Dp, Fe_R being defined as that which was complexed by 20 μM NN at pH 6.9 in the presence of 1.8 mM H₂O₂ and 5 ppm sodium dodecyl sulphate. A 30 min reaction time was allowed for Dp to react with iron(II) in seawater prior to the determination of reactive iron(III) using the same conditions as used for Fe_R. Detection limits of 0.08 nM, 0.077 nM and 0.12 nM were obtained for Fe_R, iron(III) and iron(II), respectively, using a 60 s deposition time.The method was utilised to determine the redox speciation of iron in the northern North Sea. Concentrations of Fe_R ranged between 0.8 and 3.5 nM with nutrient-like depth profiles. Iron(II) was found to be present at concentrations up to 1.2 nM, the highest concentrations occurring in the upper 20 m of the water column.     

Risk of dike failure due to flow slides

Flow slides may affect the stability of dikes. A flow slide is an instability of a submerged slope caused by liquefaction of loose, (medium) fine sand. Whether a flow slide will occur depends on the properties of the sand, which are a function of its density, and the geometry of the slope, as determined by wave and current induced scour and sedimentation. The influence of sand properties and the geometry parameters on the risk of flow slides are discussed. The application of a flow slide prediction method to an example and a risk analysis is briefly discussed.     

Circulation

Low-frequency current and temperature variability on the southeast US continental shelf during summer conditions of weak wind forcing and vertical stratification was found to be similar in many aspects to previous findings for winter, when stronger wind forcing and vertical homogeneity prevails. Subtidal variability in the outer shelf is dominated by the weekly occurrence of Gulf Stream frontal eddies and meanders. These baroclinic events strongly affect the balance of momentum in the outer shelf, but not at mid-shelf. A negative alongshore sea level slope of order −10⁻⁷ is required to balance mean along-shelf momentum at the shelf edge, similar to oceanic estimates, and can contribute to the observed northward mean flow over the shelf.Low-frequency flow at mid-shelf and coastal sea level fluctuations appear to occur as a forced wave response to local alongshore wind stress events that are coherent over the shelf domain. Momentum balances indicate a trapped wave response similar to the arrested topographic wave found in the mid-Atlantic Bight (CSANADY, 1978). Density driven currents from river discharge do not appear to be significant at mid-shelf. Cold, subsurface intrusions of deeper, nutrient rich Gulf Stream waters can occasionally penetrate to mid- and inner-shelf regions north of Cape Canaveral, causing strong phytoplankton and zooplankton responses. These events were observed following the simultaneous occurrence of upwellings from northward winds and Gulf Stream frontal eddies at the shelf break during periods when the Stream was in an onshore position. Subsurface Gulf Stream intrusions to mid-shelf occur only during the summer, when the shelf is vertically stratified and cross-shelf density gradients do not present a barrier as in winter.