Metabolic responses of Euglena gracilis under photoheterotrophic and heterotrophic conditions.

The protist Euglena gracilis has various trophic modes including heterotrophy and photoheterotrophy. To investigate how cultivation mode influences metabolic regulation, the chemical composition of cellular metabolites of Euglena gracilis grown under heterotrophic and photoheterotrophic conditions was monitored from the early exponential phase to the mid-stationary phase using two different techniques, i.e, nuclear magnetic resonance (NMR) spectroscopy and high-resolution mass spectrometry (HRMS). The combined metabolomics approach allowed an in-depth understanding of the mechanism of photoheterotrophic and heterotrophic growth for biomolecule production. Heterotrophic conditions promoted the production of polar amino and oxygenated compounds such as proteins and polyphenol compounds, especially at the end of the exponential phase while photoheterotrophic cells enhanced the production of organoheterocyclic compounds, carbohydrates, and alkaloids.

Molecular changes in phenolic compounds in Euglena gracilis cells grown under metal stress.

Metal presence in the aquatic ecosystem has increased and diversified over the last decades due to anthropogenic sources. These contaminants cause abiotic stress on living organisms that lead to the production of oxidizing molecules. Phenolic compounds are part of the defense mechanisms countering metal toxicity. In this study, the production of phenolic compounds by Euglena gracilis under three different metal stressors (i.e. cadmium, copper, or cobalt) at sub-lethal concentration was assessed using an untargeted metabolomic approach by mass spectrometry combined with neuronal network analysis (i.e. Cytoscape). The metal stress had a greater impact on molecular diversity than on the number of phenolic compounds. The prevalence of sulfur- and nitrogen-rich phenolic compounds were found in Cd- and Cu-amended cultures. Together these results confirm the impact of metallic stress on phenolic compounds production, which could be utilized to assess the metal contamination in natural waters.

Degradation pathways for organic matter of terrestrial origin are widespread and expressed in Arctic Ocean microbiomes.

BACKGROUND: The Arctic Ocean receives massive freshwater input and a correspondingly large amount of humic-rich organic matter of terrestrial origin. Global warming, permafrost melt, and a changing hydrological cycle will contribute to an intensification of terrestrial organic matter release to the Arctic Ocean. Although considered recalcitrant to degradation due to complex aromatic structures, humic substances can serve as substrate for microbial growth in terrestrial environments. However, the capacity of marine microbiomes to process aromatic-rich humic substances, and how this processing may contribute to carbon and nutrient cycling in a changing Arctic Ocean, is relatively unexplored. Here, we used a combination of metagenomics and metatranscriptomics to assess the prevalence and diversity of metabolic pathways and bacterial taxa involved in aromatic compound degradation in the salinity-stratified summer waters of the Canada Basin in the western Arctic Ocean. RESULTS: Community-scale meta-omics profiling revealed that 22 complete pathways for processing aromatic compounds were present and expressed in the Canada Basin, including those for aromatic ring fission and upstream funneling pathways to access diverse aromatic compounds of terrestrial origin. A phylogenetically diverse set of functional marker genes and transcripts were associated with fluorescent dissolved organic matter, a component of which is of terrestrial origin. Pathways were common throughout global ocean microbiomes but were more abundant in the Canada Basin. Genome-resolved analyses identified 12 clades of Alphaproteobacteria, including Rhodospirillales, as central contributors to aromatic compound processing. These genomes were mostly restricted in their biogeographical distribution to the Arctic Ocean and were enriched in aromatic compound processing genes compared to their closest relatives from other oceans. CONCLUSION: Overall, the detection of a phylogenetically diverse set of genes and transcripts implicated in aromatic compound processing supports the view that Arctic Ocean microbiomes have the capacity to metabolize humic substances of terrestrial origin. In addition, the demonstration that bacterial genomes replete with aromatic compound degradation genes exhibit a limited distribution outside of the Arctic Ocean suggests that processing humic substances is an adaptive trait of the Arctic Ocean microbiome. Future increases in terrestrial organic matter input to the Arctic Ocean may increase the prominence of aromatic compound processing bacteria and their contribution to Arctic carbon and nutrient cycles. Video Abstract.

Large-Scale Interlaboratory DI-FT-ICR MS Comparability Study Employing Various Systems.

Ultrahigh resolution mass spectrometry (UHR-MS) coupled with direct infusion (DI) electrospray ionization offers a fast solution for accurate untargeted profiling. Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometers have been shown to produce a wealth of insights into complex chemical systems because they enable unambiguous molecular formula assignment even if the vast majority of signals is of unknown identity. Interlaboratory comparisons are required to apply this type of instrumentation in quality control (for food industry or pharmaceuticals), large-scale environmental studies, or clinical diagnostics. Extended comparisons employing different FT-ICR MS instruments with qualitative direct infusion analysis are scarce since the majority of detected compounds cannot be quantified. The extent to which observations can be reproduced by different laboratories remains unknown. We set up a preliminary study which encompassed a set of 17 laboratories around the globe, diverse in instrumental characteristics and applications, to analyze the same sets of extracts from commercially available standard human blood plasma and Standard Reference Material (SRM) for blood plasma (SRM1950), which were delivered at different dilutions or spiked with different concentrations of pesticides. The aim of this study was to assess the extent to which the outputs of differently tuned FT-ICR mass spectrometers, with different technical specifications, are comparable for setting the frames of a future DI-FT-ICR MS ring trial. We concluded that a cluster of five laboratories, with diverse instrumental characteristics, showed comparable and representative performance across all experiments, setting a reference to be used in a future ring trial on blood plasma.

Influence of Carbon Sources on the Phenolic Compound Production by Euglena gracilis Using an Untargeted Metabolomic Approach.

Industrial development and urbanization has led to the diverse presence of metals in wastewater that are often improperly treated. The microalgae Euglena gracilis can tolerate high concentrations of metal via the excretion of organic metabolites, including phenolics. This study aims to evaluate how carbon amendment stimulates phenolic compound production by E. gracilis. The number, relative intensity and molecular composition of the phenolic compounds were significantly different between each of four carbon amended cultures (i.e., glutamic acid, malic acid, glucose, reduced glutathione) during the log phase. Phenolic compounds were mainly produced during the minimum growth rate, likely a response to stressful conditions. A better understanding of phenolic compounds production by E. gracilis and the impact of growth conditions will help identify conditions that favor certain phenolic compounds for dietary and metal chelation applications.

Using chemometric models to predict the biosorption of low levels of dysprosium by Euglena gracilis.

The critical rare earth element dysprosium (Dy) is integral for sustainable technologies. What is concerning is that Dy is in imminent short supply and no current replacements yet exist, coupled with increasing environmental Dy levels influenced by anthropogenic activities. This study applies chemometric methods such as response surface methodology and artificial neural networks to predict low Dy removal levels using the biosorbent Euglena gracilis. A three-factor Box-Behnken experimental design was conducted with initial concentration (1 to 100 µg L-1), contact time (30 to 180 min), and pH (3 to 8) as the three independent variables, and percentage removal and sorption capacity (q) as dependent variables. Using Dy percentage removal as response, for the worst and best conditions ranged from 0 to 92% respectively, with an average removal of 66 ± 4%. Using sorption capacity (q) as a different response variable, q varied from 0 to 93 µg/g with 27 ± 4 µg/g capacity as average. Maximum removal was 92% (q = 93 µg/g) was at pH 3, a contact time of 105 min and at a concentration of 100 µg/L. Using sorption capacity as the response variable for ANOVA, pH and metal concentrations were statistically significant factors, with lower pH and higher metal concentration having improved Dy removal, with a desirability near 1. Statistical tests such as analysis of variance, lack-of-fit, and coefficient of determination (R2) confirmed model validity. A 3-10-1 ANN network array was used to model experimental responses (q). RSM and ANN effectively modeled Dy biosorption. E. gracilis proved to be a cheap and effective biosorbent for Dy biosorption and has the potential to remediate acid mine drainage areas exhibiting low Dy concentrations.

Molecular characterization of water extractable Euglena gracilis cellular material composition using asymmetrical flow field-flow fractionation and high-resolution mass spectrometry.

Asymmetrical flow field-flow fractionation (AF4) and high-resolution Orbitrap mass spectrometry (HRMS) were used to separate and characterize cellular fractions of the dark- and light-grown Euglena gracilis cellular material. Biological replicates analyzed by HRMS shared 21-73% of commonly detected m/z values. Greater variability in shared features was found in light-grown cellular fractions (p < 0.05), likely due to small variations in growth stage. Significant differences in molecular composition were observed between AF4 cellular fractions, with dark cell fractions showing a propensity towards carbohydrate-like and tannin-like compounds, and higher double-bond equivalent (DBE) and modified aromatic index (AImod) were associated with light-grown cell fractions. Fractionation and high-resolution mass spectrometry aided characterization demonstrated the power of the AF4 to selectively cater to certain compounds/cellular entities with distinct compositional classes and double-bond equivalents and aromaticity index characteristics. Graphical abstract.

An estimation of sulfur concentrations released by three algae (Chlorella vulgaris, Chlamydomonas reinhardtii, Scenedesmus obliquus) in response to variable growth photoperiods.

In this study, we estimated extracellular concentrations of algal-derived sulfur species in response to changing photoperiods. Cultures from three algal species (Chlorella vulgaris, Chlamydomonas reinhardtii, Scenedesmus obliquus) were subjected to five different light:dark cycles (12:12, 14:10, 16:8, 18:6, 20:4 h) for a period of 3 days. Sulfur compounds including total reactive thiol concentrations, electroactive reduced sulfur species (RSS), and thiol isomers were measured using qBBr fluorescence, differential pulse cathodic stripping voltammetry (DP-CSV), and high-resolution mass spectrometry (HRMS), respectively. Total reactive thiol concentrations were greater in Scenedesmus than in Chlamydomonas and Chlorella at low light regimes (i.e., 12:12 h) whereas Chlamydomonas produced more RSS than the other two species (p < 0.05) at any light regime. Scenedesmus was the only responsive species to produce maximal electroactive RSS, and HRMS equivalent thiol compounds under low light regime, congruent with previous studies. Principal component analysis revealed relationships between qBBr-equivalent thiol and GSH-equivalent RSS concentrations for Scenedesmus and Chlamydomonas (p < 0.05) suggesting that thiols were the dominant species in the pool of electroactive RSS for these two algal species. Overall, these results showed that the light growth conditions greatly influenced the production of S-rich compounds by algae, affecting the complexation of metals such as mercury and cadmium, especially during planktonic blooms.

Aerobic and Anaerobic Bacterial Mercury Uptake is Driven by Algal Organic Matter Composition and Molecular Weight.

The biological mobilization of mercury (Hg) into microbes capable of Hg methylation is one of the limiting steps in the formation of the neurotoxin methylmercury (MeHg). Although algal dissolved organic matter (DOM) has been associated with increased MeHg production, the relationship between bacterial Hg uptake and algal DOM remains unexplored. In this study, we aimed to address how the quantity and quality of DOM, freshly harvested from several algae, affected the bacterial uptake of Hg with the use of a biosensor capable of functioning both aerobically and anaerobically. We combined biosensor measurements with high-resolution mass spectrometry and field-flow fractionation to elucidate how DOM composition and molecular weight influenced microbial Hg uptake. We showed that freshly harvested DOM from Chlorophyte and Euglena mutabilis strongly inhibited aerobic and anaerobic Hg uptake, whereas DOM harvested from Euglena gracilis did not exhibit this same pronounced effect. Once fractionated, we found that amino acids and polyamines, most abundant in Euglena gracilis DOM, were positively correlated to increase Hg uptake, suggesting that these molecules are potentially underappreciated ligands affecting Hg bioavailability. As water quality is affected by eutrophication, algal community assemblages will change, leading to variations in the nature of autochthonous DOM released in aquatic systems. Our results highlight that variations in the emergent properties of DOM originating from varying algal species can have a profound effect on bacterial Hg uptake and thus methylation.

Genomic evidence for the degradation of terrestrial organic matter by pelagic Arctic Ocean Chloroflexi bacteria.

The Arctic Ocean currently receives a large supply of global river discharge and terrestrial dissolved organic matter. Moreover, an increase in freshwater runoff and riverine transport of organic matter to the Arctic Ocean is a predicted consequence of thawing permafrost and increased precipitation. The fate of the terrestrial humic-rich organic material and its impact on the marine carbon cycle are largely unknown. Here, a metagenomic survey of the Canada Basin in the Western Arctic Ocean showed that pelagic Chloroflexi from the Arctic Ocean are replete with aromatic compound degradation genes, acquired in part by lateral transfer from terrestrial bacteria. Our results imply marine Chloroflexi have the capacity to use terrestrial organic matter and that their role in the carbon cycle may increase with the changing hydrological cycle.

Rates and processes affecting As speciation and mobility in lake sediments during aging.

Sediments from an arsenic (As) contaminated groundwater vent site were used to investigate As(III) binding, transformation and redistribution in native and iron oxide amended lake sediments using aging spiked batch reactions and a sequential extraction procedure that maintains As(V) and As(III) speciation. In the native sediments, fractionation analysis revealed that 10% of the spiked As(III) remained intact after a 32-day aging experiment and was predominantly adsorbed to the strongly sorbed (NH4H2PO4 extractable) and amorphous Fe oxide bound (H3PO4 extractable) fractions. Kinetic modelling of the experimental results allowed identifying the dominant reaction path for depletion of dissolved As(III) to As(III) absorbed on to the solid phase, followed by oxidation in the solid phase. Arsenite was initially adsorbed primarily to the easily exchangeable fraction ((NH4)2SO4 extractable), then rapidly transformed into As(V) and redistributed to the strongly sorbed and amorphous Fe oxide bound fractions. Oxidation of As(III) in recalcitrant fractions was less efficient. The iron oxide amendments illustrated the controls that iron oxides can have on As(III) binding and transformation rates. In goethite amended samples As(III) oxidation was faster and primarily occurred in the strongly sorbed and amorphous Fe oxide bound fractions. In these samples, 19.3µg Mn was redistributed (compared to the native sediment) from the easily exchangeable and crystalline Fe oxide bound fractions to the strongly sorbed and amorphous Fe oxide bound fractions, indicating that goethite may act as a catalyst for Mn(II) oxidation, thereby producing sorbed Mn(III/IV), which then appears to be involved in rapidly oxidizing As(III).

Metal (Pb, Cd, and Zn) Binding to Diverse Organic Matter Samples and Implications for Speciation Modeling.

This study evaluated the influence of dissolved organic matter (DOM) properties on the speciation of Pb, Zn, and Cd. A total of six DOM samples were categorized into autochthonous and allochthonous sources based on their absorbance and fluorescence properties. The concentration of free metal ions ( CM2+) measured by titration using the absence of gradients and Nernstian equilibrium stripping (AGNES) method was compared with that predicted by the Windermere humic aqueous model (WHAM). At the same binding condition (pH, dissolved organic carbon, ionic strength, and total metal concentration) the allochthonous DOM showed a higher level of Pb binding than the autochthonous DOM (84- to 504-fold CPb2+ variation). This dependency, however, was less pronounced for Zn (12- to 74-fold CZn2+ variation) and least for Cd (2- to 14-fold CCd2+ variation). The WHAM performance was affected by source variation through the active DOM fraction ( F). The commonly used F = 1.3 provided reliable CPb2+ for allochthonous DOMs and acceptable CCd2+ for all DOM, but it significantly under-predicted CPb2+ and CZn2+ for autochthonous DOM. Adjusting F improved CM2+ predictions, but the optimum F values were metal-specific (e.g., 0.03-1.9 for Pb), as shown by linear correlations with specific optical indexes. The results indicate a potential to improve WHAM by incorporating rapid measurement of DOM optical properties for site-specific F.

Human perturbation increases the fluxes of dissolved molybdenum from land to ocean - The case of the Jiulong River in China.

Rivers contribute a substantial amount of trace metals including molybdenum (Mo) into the oceans. The driving forces controlling the riverine fluxes of dissolved metals still remain not fully understood. Our study then investigated the spatial variations of dissolved metals including molybdenum in a typically human perturbed river, the Jiulong River (JR), China. The aim of the study is to elucidate the relevance of anthropogenic perturbation on the fluxes of dissolved metals such as molybdenum from land to ocean. Our study shows a large spatial variability of dissolved Mo across tributary to main stream of the JR. Particularly, dissolved Mo was generally low (average: 5 ± 1 nM) in the "pristine" JR headwaters, and elevated (19 ± 6 nM) along the lower river continuum. Sporadically high levels of dissolved Mo occurred in the upper North River (77 ± 19 nM), as a result of mining activities locally. Significant correlations of dissolved Mo with total dissolved solids (TDS) and dissolved strontium (Sr) were observed in the whole JR (Mo = 1.4* TDS -1.7, R2 = 0.86, p < .01; Mo = 1.2*Sr - 2.2, R2 = 0.70, p < .01, logarithmic scales). This indicates that dissolved Mo is mobilized mainly along with other major ions such as Sr during similar mineral dissolution processes. From the "pristine" headwaters to the mouth of the JR, riverine Mo fluxes at the mouth of the JR has elevated by at least 3 times due to human perturbation. Compiled historic data regarding metal fluxes from world rivers further confirmed that small and medium rivers are relatively more sensitive to human perturbation.

Influence of dissolved organic matter on dissolved vanadium speciation in the Churchill River estuary (Manitoba, Canada).

Diffusive gradients in thin films (DGT) devices were used to investigate the temporal and spatial changes in vanadium (V) speciation in the Churchill estuary system (Manitoba). Thirty-six DGT sets and 95 discrete water samples were collected at 8 river and 3 estuary sites during spring freshet and summer base flow. Dissolved V concentration in the Churchill River at summer base flow was approximately 5 times higher than those during the spring high flow (27.3 ± 18.9 nM vs 4.8 ± 3.5 nM). DGT-labile V showed an opposite trend with greater values found during the spring high flow (2.6 ± 1.8 nM vs 1.4 ± 0.3 nM). Parallel factor analysis (PARAFAC) conducted on 95 excitation-emission matrix spectra validated four humic-like (C1C4) and one protein-like (C5) fluorescent components. Significant positive relationship was found between protein-like DOM and DGT-labile V (r = 0.53, p < 0.05), indicating that protein-like DOM possibly affected the DGT-labile V concentration in Churchill River. Sediment leachates were enriched in DGT-labile V and protein-like DOM, which can be readily released when river sediment began to thaw during spring freshet.

Advanced Residuals Analysis for Determining the Number of PARAFAC Components in Dissolved Organic Matter.

Parallel factor analysis (PARAFAC) has facilitated an explosion in research connecting the fluorescence properties of dissolved organic matter (DOM) to its functions and biogeochemical cycling in natural and engineered systems. However, the validation of robust PARAFAC models using split-half analysis requires an oft unrealistically large number (hundreds to thousands) of excitation-emission matrices (EEMs), and models with too few components may not adequately describe differences between DOM. This study used self-organizing maps (SOM) and comparing changes in residuals with the effects of adding components to estimate the number of PARAFAC components in DOM from two data sets: MS (110 EEMs from nine leaf leachates and headwaters) and LR (64 EEMs from the Lena River). Clustering by SOM demonstrated that peaks clearly persisted in model residuals after validation by split-half analysis. Plotting the changes to residuals was an effective method for visualizing the removal of fluorophore-like fluorescence caused by increasing the number of PARAFAC components. Extracting additional PARAFAC components via residuals analysis increased the proportion of correctly identified size-fractionated leaf leachates from 56.0 ± 0.8 to 75.2 ± 0.9%, and from 51.7 ± 1.4 to 92.9 ± 0.0% for whole leachates. Model overfitting was assessed by considering the correlations between components, and their distributions amongst samples. Advanced residuals analysis improved the ability of PARAFAC to resolve the variation in DOM fluorescence, and presents an enhanced validation approach for assessing the number of components that can be used to supplement the potentially misleading results of split-half analysis.

Molecular characterization of phytoplankton dissolved organic matter (DOM) and sulfur components using high resolution Orbitrap mass spectrometry.

Orbitrap high resolution mass spectrometry (HRMS) with electrospray ionization in both positive and negative polarity was conducted on Suwannee River fulvic acid (SRFA), Pony Lake fulvic acid (PLFA) standards, and dissolved organic matter (DOM) released by freshwater phytoplankton (Scenedesmus obliquus, Euglena mutabilis, and Euglena gracilis). Three-dimensional van Krevelen diagrams expressing various oxygenation states of sulfur molecules and abundance plots of sulfur-containing species were constructed. Orbitrap HRMS analysis of SRFA found a high density of peaks in the lignin region (77 %) and low density of protein material (6.53 %), whereas for PLFA, 25 % of the total peaks were lignin related compared to 56 % of peaks in protein regions, comparable with other HRMS studies. Phytoplankton-derived DOM of S. obliquus, E. mutabilis, and E. gracilis was dominated by protein molecules at respective percentages of 36, 46, and 49 %, and is consistent with previous experiments examining phytoplankton-derived DOM composition. The normalized percentage of SO-containing compounds was determined among the three phytoplankton to be 56 % for Scenedesmus, 54 % for E. mutabilis, and 47 % for E. gracilis, suggesting variation between sulfur content in phytoplankton-derived DOM and differences in metal binding capacities. These results suggest the level of resolution by Orbitrap mass spectrometry is sufficient for preliminary characterization of phytoplankton DOM at an affordable cost relative to other HRMS techniques.

Examining concentrations and molecular weights of thiols in microorganism cultures and in Churchill River (Manitoba) using a fluorescent-labeling method coupled to asymmetrical flow field-flow fractionation.

In this study, molecular weights of thiols from four laboratory cultures (Scenedesmus obliquus, Chlorella vulgaris, Euglena gracilis, and Attheya septentrionalis) and the Churchill River (Manitoba) were assessed using a fluorescent-labeling method such as monobromotrimethylammoniobimane (qBBr) and asymmetrical flow field-flow fractionation (AF4) coupled to a fluorescence detector. Concentrations of thiols in extracellular fractions ranged from 6.39 ± 3.39 to 39.2 ± 7.43 µmol g(-1), and intracellular concentrations ranged from 11.5 ± 4.52 to 41.0 ± 4.1 µmol g(-1). In addition, molecular weights (MW) of intracellular thiol ranged from 493 ± 24 to 946 ± 12 Da whereas extracellular thiol MWs varied from 443 ± 36 to 810 ± 174 Da. The novel method of combining AF4 to an on-line fluorometer and the incorporation of the thiol tag provided information regarding thiol concentration and composition of controlled and natural systems. Furthermore, the proposed methods allow for the simultaneous measurement of thiol and DOM MWs produced by microorganisms. By assessing characteristics of naturally produced thiols and lab-grown thiols, information regarding heavy metal complexation can be determined.

Characterization of biochar-derived dissolved organic matter using UV-visible absorption and excitation-emission fluorescence spectroscopies.

In recent years, biochar has become of considerable interest for a variety of environmental applications. However, the feasibility of its application is entirely dependent on its physical and chemical properties, including the characteristics of biochar-derived dissolved organic matter (DOM). The goal of this study was to assess the use of optical analysis for the purpose of characterizing biochar-derived DOM. Three different biochars (slow pyrolysis birch and maple; fast pyrolysis maple) were produced and leached in distilled water over 17d. Samples were taken on days 3, 10, 13 and 17, filtered, and analyzed for DOC content. Samples were also subjected to optical analysis using UV-visible absorption and excitation-emission matrix (EEM) fluorescence spectroscopies. EEM fluorescence data were further analyzed using parallel factor analysis (PARAFAC). Absorbance and fluorescence results were combined and examined using principal component analysis (PCA). Significant differences in the water soluble organic carbon content were observed for all biochar types. The estimated aromaticity (SUVA254) and mean molecular weight (S275-295) of biochar-derived DOM were also found to differ based on biochar type. PARAFAC analysis identified three humic-like components and one protein-like component. Distinct DOM signatures were observed for each biochar type. Transformations in biochar DOM characteristics over time were also observed. The PCA showed a clear delineation in biochar types based on their optical properties. The results of this study indicate that optical analysis may provide valuable information regarding the characteristics of biochar-derived DOM.

Impacts of microbial activity on the optical and copper-binding properties of leaf-litter leachate.

Dissolved organic matter (DOM) is a universal part of all aquatic systems that largely originates with the decay of plant and animal tissue. Its polyelectrolytic and heterogeneous characters make it an effective metal-complexing agent with highly diverse characteristics. Microbes utilize DOM as a source of nutrients and energy and their enzymatic activity may change its composition, thereby altering the bioavailability and toxicity of metals. This study investigated the impacts of microbial inoculation upon the optical and copper-binding properties of freshly produced leaf-litter leachate over 168 h. Copper speciation was measured using voltammetry, and using fluorescence quenching analysis of independent fluorophores determined using parallel factor analysis (PARAFAC). Two protein/polyphenol-like and two fulvic/humic-like components were detected. Thirty-five percent of total protein/polyphenol-like fluorescence was removed after 168-h of exposure to riverine microbes. The microbial humic-like and tryptophan-like PARAFAC components retained significantly different log K values after 168 h of incubation (p < 0.05), while their complexing capacities were similar. Using voltammetry, a sixfold increase in copper-complexing capacity (CC, from 130 to 770 µmol Cu g C(-1)) was observed over the exposure period, while the conditional binding constant (log K) decreased from 7.2 to 5.8. Overall binding parameters determined using voltammetry and fluorescence quenching were in agreement. However, the electrochemically based binding strength was significantly greater than that exhibited by any of the PARAFAC components, which may be due to the impact of non-fluorescent DOM, or differences in the concentration ranges of metals analyzed (i.e., different analytical windows). It was concluded that the microbial metabolization of maple leaf leachate has a significant impact upon DOM composition and its copper-binding characteristics.

Structural and optical characterization of dissolved organic matter from the lower Athabasca River, Canada.

Dissolved organic matter (DOM) is a ubiquitous constituent of natural waters and is comprised of a variety of chemically heterogeneous molecular structures and functional groups. DOM is often considered to be a major ligand for metals in most natural waters and its reactivity is thought to be strongly dependent on its chemical composition and structure. In this study, a combination of UV/visible, emission excitation matrix fluorescence (EEM) and (1)H NMR spectroscopies were used to characterize DOM from the Athabasca River (Alberta, Canada). The chemical characterization of river DOM showed that the most upstream samples located in agricultural areas were blue-shifted and less aromatic and contained more hydrogens connected with oxygen functional groups than those in the wetland dominated area in the Athabasca oil sand deposit region. The presence of paramagnetic ions (Fe and Al) was not found to significantly affect the structural composition of DOM as revealed by (1)H NMR. Such change in the quality of DOM may have a profound impact on metal binding in the Athabasca River watershed.