Algae impacted drinking water: Does switching to chloramination produce safer drinking water?

Harmful algal (cyanobacterial) blooms (HABs) are increasing throughout the world. HABs can be a direct source of toxins in freshwater sources, and associated algal organic matter (AOM) can act as precursors for the formation of disinfection by-products (DBPs) in drinking water. This study investigated the impacts of algae on DBP formation using treatment with chloramine, which has become a popular disinfectant in the U.S. and in several other countries because it can significantly lower the levels of regulated DBPs formed. Controlled laboratory chloraminations were conducted using live field-collected algal biomass dominated by either Phormidium sp. or Microseira wollei (formerly known as Lyngbya wollei) collected from Lake Wateree and Lake Marion, SC. Sixty-six priority, unregulated or regulated DBPs were quantified using gas chromatography (GC)-mass spectrometry (MS). The presence of HAB-dominated microbial communities in source waters led to significant increases in more toxic nitrogen-containing DBPs (1.5-5 fold) relative to lake waters collected in HAB-free waters. Compared to chlorinated Phormidium-impacted waters, chloraminated waters yielded lower total DBP levels (up to 123 µg/L vs. 586 µg/L for low Br-/I- waters), but produced a greater number of brominated, iodinated, and mixed halogenated DBPs in high Br-/I- waters. Among the DBPs formed in Phormidium-impacted chloraminated waters, dichloroacetic acid, trichloromethane, chloroacetic acid, chloropropanone, and dichloroacetamide were dominant. For Microseira wollei-impacted chloraminated waters, total DBP concentrations ranged from 33 to 145 µg/L (approximately 3-5 times lower than chlorination), with dichloroacetic acid, dichloroacetamide, and trichloromethane dominant. Overall, chloramination significantly reduced calculated cytotoxicity and genotoxicity in low Br- and I- waters, but produced 1.3 fold higher calculated cytotoxicity (compared to chlorine) with high Br-/I- waters due to increased formation of more toxic iodo- and mixed halogenated DBPs.

Shoreline Drying of Microseira (Lyngbya) wollei Biomass Can Lead to the Release and Formation of Toxic Saxitoxin Analogues to the Water Column.

The harmful, filamentous cyanobacteria Microseira (Lyngbya) wollei produces several toxic analogues of saxitoxin (Lyngbya wollei toxins 1-6, or LWTs 1-6), grows in shallow water, and can deposit significant biomass on nearby shorelines. Here, we show that the LWTs are stable in the biomass during subsequent drying but that the process facilitates the later release of LWTs upon return to the water column. Under basic conditions, LWTs hydrolyzed to generate products that were significantly more neurotoxic than the initial toxins. Aqueous LWTs were subjected to conditions of covarying temperature and pH, and their degradation rates and products were determined at each condition. LWTs 1, 5, and 6 degraded faster at pH ≥ 8 at all temperatures. Their degradation products, which included decarbamoyl saxitoxin and LWT 4, were consistent with a base-catalyzed hydrolysis mechanism and represented a net increase in total biomass toxicity normalized against the equivalent toxicity of saxitoxin. The corresponding pre-exponential terms and activation energies for hydrolysis were obtained for pH 6-10 over the temperature range 10-40 °C. A locally weighted scatterplot smoothing (LOWESS) regression was developed to predict the loss of parent toxins and subsequent products in the water column under conditions corresponding to those commonly encountered in cyanobacterial blooms.

Growth of the harmful benthic cyanobacterium Microseira wollei is driven by legacy sedimentary phosphorous.

Models for cyanobacterial harmful algae blooms (cHABs) in fresh waters are usually predicated on the relationship between cyanobacterial ecology and dissolved nutrients, particularly phosphorous. Here we show legacy sediment-associated phosphorous as the primary driver of a benthic cHAB, not phosphorous in the water column. Biogeographical surveys by teams of citizen science volunteers working with the University of South Carolina identified over 200 distinct mats of Microseira wollei in Lake Wateree, SC based on toxin characterization. In sum these were estimated to affect approximately 175 km of the lake's shoreline. This growth occurred under water quality conditions that were near or below the regulatory total maximum daily load for phosphorous and nitrogen. A series of established predictive models for cyanobacterial biomass growth were applied retroactively to match the measured growth with measured water quality parameters. The only component of the system that successfully predicted microbial biomass was sedimentary phosphorous. Concentrations of the Lyngbya wollei toxins (LWTs) 1, 4, 5, and 6 were determined at multiple sites over an 18-month period and a toxin inventory for the lake was calculated. Toxin profiles between sites differed at the 95% level of confidence, establishing each site as a unique mat. An empirical model of toxin production potential based on sedimentary phosphorous was developed.

Microseira wollei and Phormidium algae more than doubles DBP concentrations and calculated toxicity in drinking water.

Warm weather and excess nutrients from agricultural runoff trigger harmful algal blooms, which can affect drinking water safety due to the presence of algal toxins and the formation of disinfection by-products (DBPs) during drinking water treatment. In this study, 66 priority, unregulated and regulated DBPs were quantified in chlorinated controlled laboratory reactions of harmful algae Microseira wollei (formerly known as Lyngbya wollei) and Phormidium using gas chromatography (GC)-mass spectrometry (MS). Live algae samples collected from algae-impacted lakes in South Carolina were chlorinated in both ultrapure water and real source waters containing natural organic matter. DBPs were also measured in finished water from a real drinking water plant impacted by a Microseira bloom. Results show that the presence of Microseira and Phormidium more than doubles total concentrations of DBPs formed by chlorination, with levels up to 586 µg/L formed in natural lake waters. Toxic nitrogen-containing DBPs also more than doubled in concentration, with levels up to 36.1, 3.6, and 37.9 µg/L for haloacetamides, halonitromethanes, and haloacetonitriles, respectively. In ultrapure water, DBPs also formed up to 314 µg/L when algae was chlorinated, demonstrating their ability to serve as direct precursors for these DBPs. When environmentally relevant levels of bromide and iodide were added to chlorination reactions, total DBPs increased 144, 51, and 24% for drinking water reservoir, Lake Marion and Lake Wateree Microseira respectively and 29% for Phormidium. Iodo-DBPs, bromochloroiodomethane, chloroiodoacetic acid, bromoiodoacetic acid, and diiodoacetic acid were observed in finished water from a drinking water plant impacted by Microseira, and bromochloroiodomethane and dibromoiodomethane were observed in chlorinated ultrapure water containing algae, bromide, and iodide. Notably, total calculated cytotoxicity tripled in Microseira-impacted waters and doubled for Phormidium-impacted waters. Calculated genotoxicity doubled for Microseira-impacted waters and more than doubled in Phormidium-impacted waters. Haloacetonitriles were major drivers of calculated cytotoxicity in algae-impacted waters, while haloacetic acids were major drivers of calculated genotoxicity in algae-impacted waters. These results provide the most extensive assessment of DBPs formed from chlorination of algae-impacted waters and highlight potential impacts to drinking water and human health. Results from this study are particularly applicable to drinking water treatment plants that employ pre-chlorination, which can cause the release of algal organic matter (AOM) precursors to form DBPs.

Fe-catalyzed sulfide oxidation in hydrothermal plumes is a source of reactive oxygen species to the ocean.

Historically, the production of reactive oxygen species (ROS) in the ocean has been attributed to photochemical and biochemical reactions. However, hydrothermal vents emit globally significant inventories of reduced Fe and S species that should react rapidly with oxygen in bottom water and serve as a heretofore unmeasured source of ROS. Here, we show that the Fe-catalyzed oxidation of reduced sulfur species in hydrothermal vent plumes in the deep oceans supported the abiotic formation of ROS at concentrations 20 to 100 times higher than the average for photoproduced ROS in surface waters. ROS (measured as hydrogen peroxide) were determined in hydrothermal plumes and seeps during a series of Alvin dives at the North East Pacific Rise. Hydrogen peroxide inventories in emerging plumes were maintained at levels proportional to the oxygen introduced by mixing with bottom water. Fenton chemistry predicts the production of hydroxyl radical under plume conditions through the reaction of hydrogen peroxide with the abundant reduced Fe in hydrothermal plumes. A model of the hydroxyl radical fate under plume conditions supports the role of plume ROS in the alteration of refractory organic molecules in seawater. The ocean's volume circulates through hydrothermal plumes on timescales similar to the age of refractory dissolved organic carbon. Thus, plume-generated ROS can initiate reactions that may affect global ocean carbon inventories.

Emerging Lyngbya wollei toxins: A new high resolution mass spectrometry method to elucidate a potential environmental threat.

Mass spectrometric methods for the quantitative and qualitative analyses of algal biotoxins are often complicated by co-eluting compounds that present analytically as interferences. This issue is particularly critical for organic polyamines, where co-eluting materials can suppress the formation of cations during electrospray ionization. Here we present an extraction procedure designed specifically to overcome matrix-derived ion suppression of algal toxins in samples of Lyngbya wollei, a filamentous benthic algae known to produce several saxitoxin analogues. Lyngbya wollei samples were collected from a large, persistent harmful algal bloom in Lake Wateree, SC. Six known Lyngbya wollei-specific toxins (LWT1-6) were successfully resolved and quantified against saxitoxin using hydrophilic interaction liquid chromatography coupled with triple quadrupole and quadrupole time-of-flight mass spectrometry. The parent ions [M2+ - H+]+ were observed for LWTs 1-6 and the [M]2+ ion was observed for LWT5. High resolution mass spectra and unique fragmentation ions were obtained for LWTs 1-6. A dilution factor of 50 resulted in a linear calibration of saxitoxin in the algae matrix. Ion suppression was resolved by sample dilution, which led to linear, positive correlations between peak area and mass of the extracted sample (R2 > 0.96). Optimized sample extraction method and instrument parameters are presented.

Aflatoxin-Exposure of Vibrio gazogenes as a Novel System for the Generation of Aflatoxin Synthesis Inhibitors.

Aflatoxin is a mycotoxin and a secondary metabolite, and the most potent known liver carcinogen that contaminates several important crops, and represents a significant threat to public health and the economy. Available approaches reported thus far have been insufficient to eliminate this threat, and therefore provide the rational to explore novel methods for preventing aflatoxin accumulation in the environment. Many terrestrial plants and microbes that share ecological niches and encounter the aflatoxin producers have the ability to synthesize compounds that inhibit aflatoxin synthesis. However, reports of natural aflatoxin inhibitors from marine ecosystem components that do not share ecological niches with the aflatoxin producers are rare. Here, we show that a non-pathogenic marine bacterium, Vibrio gazogenes, when exposed to low non-toxic doses of aflatoxin B1, demonstrates a shift in its metabolic output and synthesizes a metabolite fraction that inhibits aflatoxin synthesis without affecting hyphal growth in the model aflatoxin producer, Aspergillus parasiticus. The molecular mass of the predominant metabolite in this fraction was also different from the known prodigiosins, which are the known antifungal secondary metabolites synthesized by this Vibrio. Gene expression analyses using RT-PCR demonstrate that this metabolite fraction inhibits aflatoxin synthesis by down-regulating the expression of early-, middle-, and late- growth stage aflatoxin genes, the aflatoxin pathway regulator, aflR and one global regulator of secondary metabolism, laeA. Our study establishes a novel system for generation of aflatoxin synthesis inhibitors, and emphasizes the potential of the under-explored Vibrio's silent genome for generating new modulators of fungal secondary metabolism.

Sediment nickel bioavailability and toxicity to estuarine crustaceans of contrasting bioturbative behaviors--an evaluation of the SEM-AVS paradigm.

Robust sediment quality criteria require chemistry and toxicity data predictive of concentrations where population/community response should occur under known geochemical conditions. Understanding kinetic and geochemical effects on toxicant bioavailability is key, and these are influenced by infaunal sediment bioturbation. This study used fine-scale sediment and porewater measurement of contrasting infaunal effects on carbon-normalized SEM-AVS to evaluate safe or potentially toxic nickel concentrations in a high-binding Spartina saltmarsh sediment (4%TOC; 35-45 µmol-S2-·g(-1)). Two crustaceans producing sharply contrasting bioturbation--the copepod Amphiascus tenuiremis and amphipod Leptocheirus plumulosus--were cultured in oxic to anoxic sediments with SEM[Ni]-AVS, TOC, porewater [Ni], and porewater DOC measured weekly. From 180 to 750 µg-Ni·g(-1) sediment, amphipod bioturbation reduced [AVS] and enhanced porewater [Ni]. Significant amphipod uptake, mortality, and growth-depression occurred at the higher sediment [Ni] even when [SEM-AVS]/foc suggested acceptable risk. Less bioturbative copepods produced higher AVS and porewater DOC but exhibited net population growth despite porewater [Ni] 1.3-1.7× their aqueous [Ni] LOEC. Copepod aqueous tests with/without dissolved organic matter showed significant aqueous DOC protection, which suggests porewater DOC attenuates sediment Ni toxicity. The SEM[Ni]-AVS relationship was predictive of acceptable risk for copepods at the important population-growth level.

Geochemical production of reactive oxygen species from biogeochemically reduced Fe.

The photochemical reduction of Fe(III) complexes to Fe(II) is a well-known initiation step for the production of reactive oxygen species (ROS) in sunlit waters. Here we show a geochemical mechanism for the same in dark environments based on the tidally driven, episodic movement of anoxic groundwaters through oxidized, Fe(III) rich sediments. Sediment samples were collected from the top 5 cm of sediment in a saline tidal creek in the estuary at Murrell's Inlet, South Carolina and characterized with respect to total Fe, acid volatile sulfides, and organic carbon content. These sediments were air-dried, resuspended in aerated solution, then exposed to aqueous sulfide at a range of concentrations chosen to replicate the conditions characteristic of a tidal cycle, beginning with low tide. No detectable ROS production occurred from this process in the dark until sulfide was added. Sulfide addition resulted in the rapid production of hydrogen peroxide, with maximum concentrations of 3.85 µM. The mechanism of hydrogen peroxide production was tested using a simplified three factor representation of the system based on hydrogen sulfide, Fe(II) and Fe(III). The resulting predictive model for maximum hydrogen peroxide agreed with measured hydrogen peroxide in field-derived samples at the 95% level of confidence, although with a persistent negative bias suggesting a minor undiscovered peroxide source in sediments.

Surface charge controls the fate of Au nanorods in saline estuaries.

This work reports the distribution of negatively charged, gold core nanoparticles in a model marine estuary as a function of time. A single dose of purified polystyrene sulfonate (PSS)-coated gold nanorods was added to a series of three replicate estuarine mesocosms to emulate an abrupt nanoparticle release event to a tidal creek of a Spartina -dominated estuary. The mesocosms contained several phases that were monitored: seawater, natural sediments, mature cordgrass, juvenile northern quahog clam, mud snails, and grass shrimp. Aqueous nanorod concentrations rose rapidly upon initial dosing and then fell to stable levels over the course of approximately 50 h, after which they remained stable for the remainder of the experiment (41 days total). The concentration of nanorods rose in all other phases during the initial phase of the experiment; however, some organisms demonstrated depuration over extended periods of time (100+ h) before removal from the dosed tanks. Clams and biofilm samples were also removed from the contaminated tanks post-exposure to monitor their depuration in pristine seawater. The highest net uptake of gold (mass normalized) occurred in the biofilm phase during the first 24 h, after which it was stable (to the 95% level of confidence) throughout the remainder of the exposure experiment. The results are compared against a previous study of positively charged nanoparticles of the same size to parameterize the role of surface charge in determining nanoparticle fate in complex aquatic environments.

Surface-functionalization effects on uptake of fluorescent polystyrene nanoparticles by model biofilms.

A study was conducted to investigate the role of nanoparticle (NP) surface functionalization/charge on their uptake by biofilms. Biofilms, bacterial colonies attached to surfaces via extracellular polymers, are effective at removing suspended nanomaterials from the aqueous phase. However, the mechanisms regulating particle uptake are unknown. Here, it was shown that the mechanism was strongly dependent on the nanoparticle surface ionization, and not the core composition of the NP. Uptake experiments were conducted using laboratory-cultured biofilms. The biofilms were incubated in the presence of fluorescent polystyrene NPs with either negatively-charged surfaces (i.e. functionalized with sulfated (SO(4) (-)-NP) or carboxylated (COO(-)-NP) groups) or positively-charged surfaces (functionalized with primary amines, Amine-P). Particles with negatively-charged sulfated surfaces associated most strongly to biofilms across all experimental conditions. Associations of positively-charged amine particles with biofilms were greatest at high ionic conditions resembling those of seawater, but were sensitive to changes in ionic strength. Sorption of COO(-)-NPs was lowest, relative to other particle types, and was not sensitive to ionic strength. The results of this study support an emerging precedent that biofilms may be an effective player in the binding and sequestration of nanoparticles in aqueous systems.

Combinatorial parameter space as an empirical tool for predicting water chemistry: Fe(II) oxidation across a watershed.

Fe(II) oxidation kinetics in surface waters are a complex function of the concentration of several dissolved species that vary geographically and temporally across watersheds. This work reports an empirical, combinatorial investigation of Fe(II) oxidation that simultaneously evaluated these variations across the pH, Fe(II), PO4³â», Cl⁻, Br(-), CO3²â», and natural organic matter (NOM) axes. The work assayed the effects of independent and dependent variables through application of a novel experimental design that varied Fe(II), PO4³â», Cl⁻, Br⁻, and CO3²â» along the pH axis. Each factor was varied across concentration ranges corresponding to the natural variation between typical fresh and salt water. The system was designed to describe the oxidation of Fe(II) that occurs when Fe(II)-rich groundwaters are mixed rapidly with oxic overlaying waters as a result of tidal movement, bioturbation, dredging, and other mixing/resuspension events. Factors and interfactor interactions were statistically evaluated to determine their importance to Fe(II) oxidation at the 95% level of confidence. Significant factors were retained and used to construct predictive numerical models of Fe(II) oxidation rates. Two models (M1 and M2) were constructed to represent the conditional endmembers of unrestricted Fe cycling (M1) and restricted Fe cycling (due to forced precipitation of Fe(III), M2). The models were challenged to predict net Fe(II) oxidation rates across a watershed (the Congaree/Santee rivers, sampled at ten different locations in South Carolina). The models were generally capable of predicting Fe(II) oxidation rates to within the 95% confidence interval, although M2 consistently overpredicted the rate relative to M1. The minimum initial Fe(II) concentration needed to observe Fe cycling is estimated based on the model output.

Short-term Fe cycling during Fe(II) oxidation: exploring joint oxidation and precipitation with a combinatorial system.

The net oxidation of Fe(II)aq by dioxygen initiates a suite of reactions including the oxidation of multiple Fe(II) complexes, generation of secondary oxidants, Fe(III) reduction, and precipitation of insoluble products. This work reports application of a multifactorial strategy to describe the oxidation of Fe(II) under conditions that correspond to those found where Fe(II)-rich groundwaters mix rapidly with overlying oxygenated waters. Response surfaces were constructed describing the relationship of the net oxidation process with mixtures of the common ligands chloride (Cl-), bromide (Br-), total carbonate (CO3(2-)), Fe(II), and Suwannee River natural organic matter (SRNOM) at pH 8.00. Response surfaces were generated in the presence and absence of added phosphate, representing conditional end members corresponding to geographical locations where Fe(III) precipitation is respectively forced and unconstrained. Comparison of net Fe(II) oxidation rates in the presence and absence of phosphate then enabled resolution of the relative contributions of Fe(II) oxidation and Fe(III) reduction to the overall process. The differences between the two surfaces demonstrated the importance of Fe(II) regeneration on the rapid (min) time scale during net oxidation. The minimum Fe(II) concentration necessary to initiate measurable cycling is reported. The presence of reactive oxygen species was evaluated through the use of probes added to the center point condition of the experimental matrix. Analysis of the statistical significance of the Fe(II)-factor relationships demonstrated that over the conditional scale of the experiments complexation of Fe(II) by the selected ligands did not correlate to the experimental outcome.

Reaction of N-acylhomoserine lactones with hydroxyl radicals: rates, products, and effects on signaling activity.

Chemical communication in bacteria, sometimes called quorum sensing, is a fundamental microbial process that is based on the exchange of molecular signals between cells. The signaling molecules involved in this process are thermodynamically unstable in some environments and their degradation affects microbial communication. This work reports the oxidation of a series of substituted N-acylhomoserine lactones (AHLs, a class of quorum sensing signals) by hydroxyl radicals. The corresponding bimolecular rate constants were obtained and correlated positively with the length of the acyl side chain (C, in numbers of carbon atoms) ranging from 2.4 × 10(9) M(-1) s(-1) to 9.4 × 10(9) M(-1) s(-1) (C4- to C10-AHL), 2.4 × 10(8) M(-1) s(-1) for 3-oxo C6-AHL, and 2.94 × 10(9) M(-1) s(-1) for 3-oxo C8-AHL. Liquid chromatography-mass spectrometric techniques were applied to qualify the identity and quantify the yields of the hydroxyl radical oxidation products of C6-AHL (aldo, keto, and hydroxylated C6-analogues identified). The biological activity of C6-AHL and associated products was determined using the Vibrio harveyi bioluminescence bioassay. Oxidation resulted in a net increase in assay response indexed against the starting AHL. This result suggested that the application of HO• based technologies such as advanced oxidation processes for biofilm control may result in unintended quorum sensing responses by microbial communities.

Multivariate examination of Fe(II)/Fe(III) cycling and consequent hydroxyl radical generation.

The introduction of Fe(II)(aq) into aerated solutions resulted in net Fe(II) oxidation with concomitant, rapid Fe(II)/Fe(IIII) cycling and concurrent generation of reactive oxygen species. The effect of mixtures of naturally occurring solutes on Fe(II)/Fe(III) cycling and the concurrent oxidation of dissolved organics is reported. The experimental strategy was based on a multivariate, microscale, high-throughput approach for evaluating the effect of covarying concentrations of bromide, iodide, Suwannee River natural organic matter (SRNOM), chloride, and total carbonate species. Superoxide and HO• were evaluated at the center point condition of the experimental design with selective scavengers (superoxide dismutase and benzoic acid). The rate of Fe(II) oxidation decreased in the presence of these scavengers, indicating it is a function of oxygen, superoxide, and HO•. HO• generated during Fe(II)/Fe(III) cycling was quantified with the selective probe 1,3-dicyanotetrachlorobenzene (DCTCB). Through the range of experimental conditions of this design, the ratio of the number of moles of HO• produced to the number of moles of Fe(II) consumed varied from 3 to 750, corresponding to approximately 10 to 2200 Fe(II)/Fe(III) cycles, respectively. The implications of these findings with respect to organic oxidation during the aeration of anoxic Fe(II) rich groundwaters are discussed.

Transfer of gold nanoparticles from the water column to the estuarine food web.

Within the next five years the manufacture of large quantities of nanomaterials may lead to unintended contamination of terrestrial and aquatic ecosystems. The unique physical, chemical and electronic properties of nanomaterials allow new modes of interaction with environmental systems that can have unexpected impacts. Here, we show that gold nanorods can readily pass from the water column to the marine food web in three laboratory-constructed estuarine mesocosms containing sea water, sediment, sea grass, microbes, biofilms, snails, clams, shrimp and fish. A single dose of gold nanorods (65 nm length x 15 nm diameter) was added to each mesocosm and their distribution in the aqueous and sediment phases monitored over 12 days. Nanorods partitioned between biofilms, sediments, plants, animals and sea water with a recovery of 84.4%. Clams and biofilms accumulated the most nanoparticles on a per mass basis, suggesting that gold nanorods can readily pass from the water column to the marine food web.

The adsorption of saxitoxin to clays and sediments in fresh and saline waters.

The adsorption of saxitoxin to Na- and Ca-montmorillonite, kaolin (crystalline and amorphous), kaolinite, Bread and Butter Creek sediment (an estuarine tidal creek), Gulf of Mexico sediment, and Santa Barbara Basin sediment in deionized water and 32 per thousand salinity simulated seawater (Instant Ocean) is reported. Adsorption was partially reversible for all cases and best described using a Freundlich isotherm. The corresponding Freundlich constants (K(F)) ranged from 8.83 x 10(3)micromol/kg to 6.76 x 10(4)micromol/kg for freshwater and 4.73 x 10(3)micromol/kg-1.11 x 10(4)micromol/kg for seawater. There is a positive linear correlation seen between the K(F) values and the cation-exchange capacity of the adsorbents. The release of saxitoxin from previously equilibrated adsorbents was determined in freshwater (0-18%) and seawater (4-53%).

Identification of hydroxyl radical oxidation products of N-hexanoyl-homoserine lactone by reversed-phase high-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry.

A reversed-phase high-performance liquid chromatography/electrospray tandem mass spectrometry method was developed for the characterization of hydroxyl radical oxidation products of N-hexanoyl-homoserine lactone (C6-HSL), a member of the N-acylhomoserine lactone (AHL) class of microbial quorum-sensing signaling molecules identified in many Gram-negative strains of bacteria. Six products were identified: four with molecular weight (MW) of 213 and two with MW of 260. The characteristic product ions formed through collision-induced dissociation (CID) provided diagnostic structural information. One of the photolysis products was determined to be N-(3-oxohexanoyl)homoserine lactone (3OC6-HSL), a highly active quorum-sensing signal, by comparison with a reference standard. Three structural isomers with the same mass as 3OC6-HSL were identified as acyl side chain oxidized C6-HSL (keto/enol functionalized) by accurate mass measurement and the structures of these products were proposed from CID spectral interpretation. Two structural isomers formed from concurrent oxidation and nitration of C6-HSL were also observed and their structures were postulated based on CID spectra. In addition to the six hydroxyl radical oxidation products formed from the C6-HSL precursor, five additional compounds generated from combined oxidation and lactonolysis of C6-HSL were identified and structures were postulated.

Use of multiparametric techniques to quantify the effects of naturally occurring ligands on the kinetics of Fe(II) oxidation.

A multifactorial experimental design was employed to quantify and rank the effects of a series of ligands on the rate of Fe(II) (18 microM) oxidation in a system containing chloride, sulfate, carbonate/bicarbonate, fluoride, and natural organic matter (NOM) at pH 8.34 +/- 0.13. Several factors and combinations thereof correlated with the rate of Fe(II) oxidation at the 95% level of confidence. Presented in decreasing order of significance, those factors were carbonate/bicarbonate, NOM, sulfate, chloride, the sulfate/fluoride interaction, and fluoride. The center point of the experimental design was repeated with different organic matters substituted, including Nordic Reservoir NOM, fulvic and humic acids; Suwannee River NOM, fulvic and humic acids; and Pony Lake fulvic acid. Despite the widely differing geographical origins of these organic materials, their overall impact on the oxidation rate of Fe(II) was consistent with the observed rate varying no more than a factor of 2 as a function of different organic matters (on a milligrams of carbon per liter basis). The utility of the pentafactorial response surface model (based on Nordic Lake NOM) to predict Fe(II) oxidation rates was evaluated for different natural water samples, including two seawater and one freshwater.