Description: Copper, Cd and Zn can be found at elevated concentrations in contaminated estuarine and coastal waters and have potential toxic effects on phytoplankton species. In this study, the effects of these metals on the intracellular production of the polypeptides phytochelatin and glutathione by the marine diatom Phaeodactylum tricornutum were examined in laboratory cultures. Single additions of Cu and Cd (0.4 μM Cu2 and 0.45 μM Cd2+) to the culture medium induced the production of short-chained phytochelatins ((γ-Glu-Cys) n -Gly where n = 2-5), whereas a single addition of Zn (2.2 μM Zn2+) did not stimulate phytochelatin production. Combination of Zn with Cu resulted in a similar phytochelatin production compared with a single Cu addition. The simultaneous exposure to Zn and Cd led to an antagonistic effect on phytochelatin production, which was probably caused by metal competition for cellular binding sites. Glutathione concentrations were affected only upon exposure to Cd (85 increase) or the combination of Cd with Zn (65 decrease), relative to the control experiment. Ratios of phytochelatins to glutathione indicated a pronounced metal stress in response to exposures to Cu or Cd combined with Zn. This study indicates that variabilities in phytochelatin and glutathione production in the field can be explained in part by metal competition for cellular binding sites. © Springer 2006.

Description: Phytochelatins (PCs) and glutathione (GSH) are -SH-containing compounds produced by a range of organisms for intracellular functions, such as protection against oxidative stress, metal detoxification and regulation of intracellular metal concentrations. These compounds, particularly PCs, have a potential use as metal-stress indicators for phytoplankton in natural waters. Despite their important roles, there is a paucity of data on intracellular GSH and PCs produced by natural phytoplankton assemblages. Current analytical methods for the determination of these compounds in phytoplankton from natural waters are based on high-performance liquid chromatography (HPLC) with detection of fluorescent derivatives, and comprise multi-step protocols. In this article, we discuss the analytical methods for HPLC determination of PCs and GSH, as there are limitations and practical challenges when they are applied to environmental studies. © 2005 Elsevier Ltd. All rights reserved.

Description: Phytoplankton deal with metal toxicity using a variety of biochemical strategies. One of the strategies involves glutathione (GSH) and phytochelatins (PCs), which are metal-binding thiol peptides produced by eukaryotes and these compounds have been related to several intracellular functions, including metal detoxification, homeostasis, metal resistance and protection against oxidative stress. This paper assesses our state of knowledge on the production of PCs and GSH by marine phytoplankton in laboratory and field conditions and the possible applications of PCs for environmental purposes. Good relationships have been observed between metal exposure and PC production in phytoplankton in the laboratory with Cd, Pb, and Zn showing the greatest efficacy, thereby indicating that PCs have a potential for application as a biomarker. Fewer studies on PC distributions in particulate material have been undertaken in the field. These studies show that free Cu has a strong relationship with the levels of PC in the particulate material. The reason for this could be because Cu is a common contaminant in coastal waters. However it could also be due to the lack of measurements of other metals and their speciation. GSH shows a more complex relationship to metal levels both in the laboratory and in the field. This is most likely due to its multifunctionality. However, there is evidence that phytoplankton act as an important source of dissolved GSH in marine waters, which may form part of the strong organic ligands that control metal speciation, and hence metal toxicity. © 2006 by the Phycological Society of America.

Description: Sorption of trace metals by suspended particulate matter (SPM) in estuarine systems has important implications for the fate of dissolved metals in these waters. This paper describes the development of a single extraction procedure for SPM-associated trace metals, using a ligand competition approach with EDTA as the added complexing ligand. The use of EDTA allows the determination of available particulate trace metals using well defined constraints with respect to the competition for trace metals between EDTA and the particles. Incubation experiments showed that equilibrium times between EDTA and particulate material of 72h were required to reach equilibrium for most of the metals studied (Cu, Zn, Mn, Ni, Co, Al, Fe, Pb and Mg). Optimum conditions included a 0.05M EDTA concentration and the use of an extractant: particulate matter ratio of 200:1 (v:w). Kinetic calculations on data from the incubation experiments were used to calculate the apparent stability constants (K(MeS)) for the metal-particulate matter interaction and indicated values ranging from 10-2.1 for K(MgS) to 10-13.5 for K(CuS). Copyright (C) 1999 Elsevier Science B.V.

Description: Siderophores are chelates produced by bacteria as part of a highly specific iron uptake mechanism. They are thought to be important in the bacterial acquisition of iron in seawater and to influence iron biogeochemistry in the ocean. We have identified and quantified two types of siderophores in seawater samples collected from the Atlantic Ocean. These siderophores were identified as hydroxamate siderophores, both ferrioxamine species representative of the more soluble marine siderophores characterized to date. Ferrioxamine G was widely distributed in surface waters throughout the Atlantic Ocean, while ferrioxamine E had a more varied distribution. Total concentrations of the two siderophores were between 3 and 20 pM in the euphotic zone. If these compounds are fully complexed in seawater, they represent approximately 0.2-4.6 of the 〈0.2 μm iron pool. Our data confirm that siderophore-mediated iron acquisition is important for marine heterotrophic bacteria and indicate that siderophores play an important role in the oceanic biogeochemical cycling of iron. © 2008 American Chemical Society.

Description: Four surveys of the Huelva Estuary in southwest Spain and its sources, the Tinto and the Odiel Rivers, were carried out between 1996 and 1998. The surveys investigated the impact of metalliferous mining of sulfide-rich ores in the catchment area on metal speciation, metal concentrations in a macrophyte, and phytoplankton diversity and abundance. Chemical speciation measurements in the lower Tinto Estuary showed that metals were predominantly electrochemically labile (>99 of total dissolved Cu, Co, and Ni at 10 μM Cu, 424 nM Co, and 500 nM Ni, S = 28). Concentrations of Cu complexing ligands and free cupric ions Cu2+ in the Gulf of Cádiz ranged between 5.3 and 38 nM and 0.2-7.9 pM, respectively, with conditional stability constants of the ligands of log K′CuL = 11.7-12.6. At enhanced dissolved Cu concentrations in the lower Huelva Estuary, Cu complexing ligands were saturated with Cu, resulting in nanomolar Cu2+, which increased upstream. Metal tissue concentrations of the macrophyte Blindingia marginata were high, and a clear relationship between dissolved labile Cu and macrophyte tissue Cu concentrations was observed. A low biodiversity was observed in the Huelva system (Shannon-Wiener indices (H) typically 〈0.2). Nevertheless, the maximum biomass was observed in the lower Tinto Estuary, which showed high labile metal and nutrient concentrations and a low biodiversity (H 〈 0.02), thereby suggesting adaptation through evolutionary processes of the phytoplankton community to the harsh conditions. © 2007 American Chemical Society.

Description: The behaviour of a series of hydroxamate siderophores - microbially produced iron complexes - was investigated using electrospray ionisation mass spectrometry (ESI-MS). Three groups of iron hydroxamate siderophores, namely the ferrioxamines, ferrichromes and coprogens/ fusigens, were separated by high-performance liquid chromatography (HPLC) prior to ESI and MS2 fragmentation. For the majority of the siderophores, both protonated molecules and sodium adducts were observed. The most abundant ion was selected for collision-induced fragmentation. Potential fragmentation mechanisms are postulated and discussed. Fragmentation patterns differed between siderophore groups; however, common fragmentation patterns were observed for siderophore ions within the groups examined. Cleavage frequently occurred at carbon-nitrogen or carbon-oxygen bonds. Fragmentation of the ions also involved cleavage of iron-oxygen bonds and transfer of the charge to iron.

Description: Short-term iron enrichment experiments were carried out with samples collected in areas with different phytoplankton activity in the northern North Sea and northeast Atlantic Ocean in the summer of 1993. The research area was dominated by high numbers of pico-phytoplankton, up to 70,000 ml−1. Maximum chlorophyll a concentrations varied from about 1.0 μg l−1 in a high-reflectance zone (caused by loose coccoliths, remnants from a bloom of Emiliania huxleyi) and about 3.5 μg l−1 in a zone in which the phytoplankton were growing, to about 0.5 μg l−1 in the northeast Atlantic Ocean. From the high-reflectance zone to the northeast Atlantic Ocean, nitrate concentrations increased from 0.5 μM to 6.0 μM. Concentrations of reactive iron in surface water showed an opposite trend and decreased from about 2.6 nM in the high-reflectance zone to 〈1.0 nM in the northeast Atlantic Ocean. In the research area, no signs of true iron deficiency were found, but iron enrichments in the high-reflectance zone, numerically dominated by Synechococcus sp., resulted in increased nitrate uptake. Ammonium uptake was hardly affected. Strong support for the effect of Fe on cell physiology is given by the increase in the f-ratio. Net growth rates of the phytoplankton (changes in cell numbers over 24 h) were almost unchanged. Phytoplankton collected from the northeast Atlantic Ocean, did not show changes in the nitrogen metabolism upon addition of iron. Net growth rates in these incubations were low or negative, with only slightly higher values with additional iron.

Description: Eight techniques for cleaning particulate material and epibionts from the tissue of Fucus vesiculosus were tested on seaweed collected from a metal-contaminated estuary in the southwest of England. When assessed by scanning electron microscopy, only one was found to have efficiently removed the particulate material on the tissue. This technique consisted of applying a 1:9 ethanol:seawater mixture to the surface of the thallus, and scraping with a PTFE spatula. Analysis of Cu, As, Fe, Mn, and Zn content in the tissues cleaned by this technique showed that, in this case, only Fe concentrations decreased significantly when compared to tissue cleaned by rinsing in seawater only. The study is the first to systematically assess cleaning techniques for marine macroalgae and illustrates the need for standard methods for the sampling and analysis of seaweed tissue

Description: Variations in the speciation of iron in the northern North Sea were investigated in an area covering at least two different water masses and an algal bloom, using a combination of techniques. Catalytic cathodic stripping voltammetry was used to measure the concentrations of reactive iron (FeR) and total iron (FeT) in unfiltered samples, while dissolved iron (FeD) was measured by GFAAS after extraction of filtered sea water. FeR was defined by the amount of iron that complexed with 20 μM 1-nitroso-2-napthol (NN) at pH 6.9. FeT was determined after UV-digestion at pH 2.4. Concentrations of natural organic iron complexing ligands and values for conditional stability constants, were determined in unfiltered samples by titration. Mean concentrations of 1.3 nM for FeR, 10.0 nM for FeT and 1.7 nM for FeD were obtained for the area sampled. FeR concentrations increased towards the south of the area investigated, as a result of the increased influence of continental run off. FeR concentrations were found to be enhanced below the nutricline (below ∼40 m) as a result of the remineralisation of organic material. Enhanced levels of FeT were observed in some surface samples and in samples collected below 30 m at stations in the south of the area studied, thought to be a result of high concentrations of biogenic particulate material and the resuspended sediments respectively. FeD concentrations varied between values similar to those of FeT in samples from the north of the area to values similar to those of FeR in the south. The bloom was thought to have influenced the distribution of both FeR and FeT, but less evidence was observed for any influence on FeR and FeD. The concentration of organic complexing ligands, which could possibly include a contribution from adsorption sites on particulate material, increased slightly in the bloom area and in North Sea waters. Iron was found to be fully (99.9%) complexed by the organic complexing ligands at a pH of 6.9 and largely complexed (82–96%) at pH 8. The ligands were almost saturated with iron suggesting that the ligand concentration could limit the concentration of iron occurring as dissolved species.