Controls on the photochemical production of hydrogen peroxide in Lake Erie.

In Lake Erie, toxin-forming harmful algal blooms (HABs) occur following high concentrations of hydrogen peroxide (H2O2). Correlation between H2O2 concentrations and HABs revealed knowledge gaps on the controls of H2O2 production in Lake Erie. One way H2O2 is produced is upon absorption of sunlight by the chromophoric fraction of dissolved organic matter (CDOM). Rates of this photochemical production of H2O2 may increase in proportion to the apparent quantum yield of H2O2 (ΦH2O2,λ) from CDOM. However, the ΦH2O2,λ for H2O2 production from CDOM remains too poorly constrained to predict the magnitude and range of photochemically produced H2O2, particularly in freshwaters like Lake Erie. To address this knowledge gap, the ΦH2O2,λ was measured approximately biweekly from June-September 2019 in the western basin of Lake Erie along with supporting analyses (e.g., CDOM concentration and composition). The average ΦH2O2,λ in Lake Erie was within previously reported ranges. However, the ΦH2O2,λ varied 5-fold in space and time. The highest ΦH2O2,λ was observed in the Maumee River, a tributary of Lake Erie. In nearshore waters of Lake Erie, the ΦH2O2,λ decreased about five-fold from June through September. Integration of the controls of photochemical production of H2O2 in Lake Erie show that the variability in rates of photochemical H2O2 production was predominantly due to the ΦH2O2,λ. In offshore waters, CDOM concentration also strongly influenced photochemical H2O2 production. Together, the results confirm prior work suggesting that photochemical production of H2O2 contributes but likely cannot account for all the H2O2 associated with HABs in Lake Erie.

Heterotrophic Bacteria Dominate Catalase Expression during Microcystis Blooms.

In the oligotrophic oceans, key autotrophs depend on "helper" bacteria to reduce oxidative stress from hydrogen peroxide (H2O2) in the extracellular environment. H2O2 is also a ubiquitous stressor in freshwaters, but the effects of H2O2 on autotrophs and their interactions with bacteria are less well understood in freshwaters. Naturally occurring H2O2 in freshwater systems is proposed to impact the proportion of microcystin-producing (toxic) and non-microcystin-producing (nontoxic) Microcystis in blooms, which influences toxin concentrations and human health impacts. However, how different strains of Microcystis respond to naturally occurring H2O2 concentrations and the microbes responsible for H2O2 decomposition in freshwater cyanobacterial blooms are unknown. To address these knowledge gaps, we used metagenomics and metatranscriptomics to track the presence and expression of genes for H2O2 decomposition by microbes during a cyanobacterial bloom in western Lake Erie in the summer of 2014. katG encodes the key enzyme for decomposing extracellular H2O2 but was absent in most Microcystis cells. katG transcript relative abundance was dominated by heterotrophic bacteria. In axenic Microcystis cultures, an H2O2 scavenger (pyruvate) significantly improved growth rates of one toxic strain while other toxic and nontoxic strains were unaffected. These results indicate that heterotrophic bacteria play a key role in H2O2 decomposition in Microcystis blooms and suggest that their activity may affect the fitness of some Microcystis strains and thus the strain composition of Microcystis blooms but not along a toxic versus nontoxic dichotomy. IMPORTANCE Cyanobacterial harmful algal blooms (CHABs) threaten freshwater ecosystems globally through the production of toxins. Toxin production by cyanobacterial species and strains during CHABs varies widely over time and space, but the ecological drivers of the succession of toxin-producing species remain unclear. Hydrogen peroxide (H2O2) is ubiquitous in natural waters, inhibits microbial growth, and may determine the relative proportions of Microcystis strains during blooms. However, the mechanisms and organismal interactions involved in H2O2 decomposition are unexplored in CHABs. This study shows that some strains of bloom-forming freshwater cyanobacteria benefit from detoxification of H2O2 by associated heterotrophic bacteria, which may impact bloom development.

Efficacy of selected pretreatment processes in the mitigation of low-pressure membrane fouling and its correlation to their removal of microbial DOM.

Membrane fouling by dissolved organic matter (DOM), especially microbially-derived DOM, is a major challenge for ultrafiltration (UF) membranes in water purification. Fouling may be mitigated by pretreating feed waters; however, there are no comprehensive studies that compare the fouling reduction efficacies across different pretreatment processes. Further, there is a limited understanding of the relationship between fouling reduction efficacy and microbially-derived DOM removal from source waters. Accordingly, the objectives of this study were to: (i) evaluate and compare the efficacies of five pretreatment processes in reducing UF membrane fouling by DOM; and (ii) investigate whether a relationship exists between membrane fouling reduction and microbially-derived DOM removal by pretreatment processes. We investigated seven water sources and a polyvinylidene fluoride hollow-fiber UF membrane using bench-scale fouling tests. Dissolved organic carbon content, ultraviolet absorbance and fluorescence excitation-emission matrix spectroscopy were used to assess DOM concentration and composition. Alum and ferric chloride coagulation were the most effective pretreatment processes in reducing membrane fouling, anion exchange was moderately effective, and PAC adsorption and chlorine pre-oxidation were the least effective. Consistent with previous studies, microbially-derived DOM was the major contributor to UF membrane fouling regardless of water source or pretreatment type. Fouling reduction was strongly correlated with the reduction of microbially-derived DOM in foulant layers but not from source waters. This result indicates that a fraction of the total microbially-derived DOM in feed waters was responsible for UF fouling. Overall, pretreatment processes that remove microbially-derived DOM are well-suited for UF membrane fouling reduction.

Direct noninvasive 1 H NMR analysis of stream water DOM: Insights into the effects of lyophilization compared with whole water.

NMR spectroscopy is widely used in the field of aquatic biogeochemistry to examine the chemical structure of dissolved organic matter (DOM). Most aquatic DOM analyzed by proton NMR (1 H NMR) is concentrated mainly by freeze-drying prior to analysis to combat low concentrations, frequently <100 µM C, and eliminate interference from water. This study examines stream water with low dissolved organic carbon content by 1 H NMR with a direct noninvasive analysis of whole water using a water-suppression technique. Surface waters, collected from the headwaters of the Rio Tempisquito, Costa Rica, were examined directly, and the spectral characteristics were compared with those of the traditional preanalysis freeze-drying approach revealing significant differences in the relative intensity of peaks between the whole water and freeze-dried DOM. The freeze-dried DOM required less time to obtain quality spectra, but several peaks were missing compared with the spectra of whole water DOM; notably the most dominant peak in the spectrum constituting roughly 10% of the DOM. The stream water DOM showed an increase in the relative intensity of aliphatic methyl and methylene groups and a decrease in carbonyl, carboxyl, and carbohydrate functionalities after freeze-drying. The results of this study show that freeze-drying alters the original composition of DOM and thus freeze-dried DOM may not represent the original DOM. The information gained from whole water analysis of stream water DOM in a noninvasive fashion outweighs the attraction of reduced analysis times for preconcentrated samples, particularly for studies interested in investigating the low molecular weight fraction of DOM.

Experimental metatranscriptomics reveals the costs and benefits of dissolved organic matter photo-alteration for freshwater microbes.

Microbes and sunlight convert terrigenous dissolved organic matter (DOM) in surface waters to greenhouse gases. Prior studies show contrasting results about how biological and photochemical processes interact to contribute to the degradation of DOM. In this study, DOM leached from the organic layer of tundra soil was exposed to natural sunlight or kept in the dark, incubated in the dark with the natural microbial community, and analysed for gene expression and DOM chemical composition. Microbial gene expression (metatranscriptomics) in light and dark treatments diverged substantially after 4 h. Gene expression suggested that sunlight exposure of DOM initially stimulated microbial growth by (i) replacing the function of enzymes that degrade higher molecular weight DOM such as enzymes for aromatic carbon degradation, oxygenation, and decarboxylation, and (ii) releasing low molecular weight compounds and inorganic nutrients from DOM. However, growth stimulation following sunlight exposure of DOM came at a cost. Sunlight depleted the pool of aromatic compounds that supported microbial growth in the dark treatment, ultimately causing slower growth in the light treatment over 5 days. These first measurements of microbial metatranscriptomic responses to photo-alteration of DOM provide a mechanistic explanation for how sunlight exposure of terrigenous DOM alters microbial processing and respiration of DOM.

Assessing the prevalence, products, and pathways of dissolved organic matter partial photo-oxidation in arctic surface waters.

In sunlit waters, photodegradation of dissolved organic matter (DOM) yields completely oxidized carbon (i.e., CO2) as well as a suite of partially oxidized compounds formed from oxygen incorporation (i.e., partial photo-oxidation). Of these two groups of DOM photo-products, more studies focus on CO2 (a greenhouse gas) than on partially oxidized DOM, which is likely a diverse group of compounds with poorly constrained roles in aquatic carbon cycling or biogeochemistry. The objective of this study is to address knowledge gaps on the prevalence, products, and pathways of DOM partial photo-oxidation. Here we traced the photochemical incorporation of isotopically labelled 18O2 into DOM isolated from Alaskan Arctic surface waters using high-resolution mass spectrometry. Complete and partial photo-oxidation of DOM was also quantified as CO2 production and O2 consumption. The majority of 18O-containing partial oxidation photo-products were classified as carboxylic rich alicyclic molecules (CRAM) and overlapped in composition with previously reported photo-products known to result from the oxidation of DOM by singlet oxygen. These results support a previously proposed hypothesis that photo-oxidation by singlet oxygen may contribute to the formation of CRAM, a compound class of DOM ubiquitously observed in surface waters. The novel application of an isotopic tracer for oxygen incorporation with a mass balance approach to quantify complete and partial photo-oxidation of DOM revealed that less than one mol of O2 is required to produce one mol of CO2. A sensitivity analysis based on this new knowledge demonstrated that the magnitude of DOM partial photo-oxidation may be underestimated by up to four-fold. Consequently, partial photo-oxidation likely plays a more prominent role in shaping DOM composition in sunlit waters of the Arctic than previously understood. Therefore, partial photo-oxidation should be increasingly incorporated into the experimental framework of studies focused on DOM composition in surface waters.

Effects of vertical hydrodynamic mixing on photomineralization of dissolved organic carbon in arctic surface waters.

Photomineralization, the transformation of dissolved organic carbon (DOC) to CO2 by sunlight, is an important source of CO2 in arctic surface waters. However, quantifying the role of photomineralization in inland waters is limited by the understanding of hydrologic controls on this process. To bridge this gap, this study evaluates mixing limitations, i.e., whether and by how much vertical mixing limits the depth-integrated photomineralization rate, in freshwater systems. We developed a conceptual model to qualitatively assess mixing limitations across the range of light attenuation and hydrologic conditions observed in freshwaters. For the common case of exponential light attenuation over depth, we developed a mathematical model to quantify mixing limitation, and used this model to assess a range of arctic freshwater systems. The results demonstrate that mixing limitations are important when there is significant light attenuation by suspended sediment (SS), which is the case in some arctic, boreal and temperate waters. Mixing limitation is pronounced when light attenuation over depth is strong and when the photomineralization rate at the water surface exceeds the vertical mixing rate. Arctic streams and rivers have strong vertical mixing relative to surface photomineralization, such that model results demonstrate no mixing limitation regardless of how much SS is present. Our analysis indicates that well-mixed assumptions used in prior work are valid in many, but not all, arctic surface waters. The effects of mixing limitations in reducing the photomineralization rate must be considered in arctic lakes with high SS concentrations.

Photochemical alteration of organic carbon draining permafrost soils shifts microbial metabolic pathways and stimulates respiration.

In sunlit waters, photochemical alteration of dissolved organic carbon (DOC) impacts the microbial respiration of DOC to CO2. This coupled photochemical and biological degradation of DOC is especially critical for carbon budgets in the Arctic, where thawing permafrost soils increase opportunities for DOC oxidation to CO2 in surface waters, thereby reinforcing global warming. Here we show how and why sunlight exposure impacts microbial respiration of DOC draining permafrost soils. Sunlight significantly increases or decreases microbial respiration of DOC depending on whether photo-alteration produces or removes molecules that native microbial communities used prior to light exposure. Using high-resolution chemical and microbial approaches, we show that rates of DOC processing by microbes are likely governed by a combination of the abundance and lability of DOC exported from land to water and produced by photochemical processes, and the capacity and timescale that microbial communities have to adapt to metabolize photo-altered DOC.The role of dissolved organic carbon (DOC) photo-alteration in the microbial respiration of DOC to CO2 is unclear. Here, the authors show that the impact of this mechanism depends on whether photo-alteration of DOC produces or removes molecules used by native microbial communities prior to light exposure.

Cyanobacterial harmful algal blooms are a biological disturbance to Western Lake Erie bacterial communities.

Human activities are causing a global proliferation of cyanobacterial harmful algal blooms (CHABs), yet we have limited understanding of how these events affect freshwater bacterial communities. Using weekly data from western Lake Erie in 2014, we investigated how the cyanobacterial community varied over space and time, and whether the bloom affected non-cyanobacterial (nc-bacterial) diversity and composition. Cyanobacterial community composition fluctuated dynamically during the bloom, but was dominated by Microcystis and Synechococcus OTUs. The bloom's progression revealed potential impacts to nc-bacterial diversity. Nc-bacterial evenness displayed linear, unimodal, or no response to algal pigment levels, depending on the taxonomic group. In addition, the bloom coincided with a large shift in nc-bacterial community composition. These shifts could be partitioned into components predicted by pH, chlorophyll a, temperature, and water mass movements. Actinobacteria OTUs showed particularly strong correlations to bloom dynamics. AcI-C OTUs became more abundant, while acI-A and acI-B OTUs declined during the bloom, providing evidence of niche partitioning at the sub-clade level. Thus, our observations in western Lake Erie support a link between CHABs and disturbances to bacterial community diversity and composition. Additionally, the short recovery of many taxa after the bloom indicates that bacterial communities may exhibit resilience to CHABs.

Minimization of short-term low-pressure membrane fouling using a magnetic ion exchange (MIEX(®)) resin.

Two challenges to low-pressure membrane (LPM) filtration are limited rejection of dissolved organic matter (DOM) and membrane fouling by DOM. The magnetic ion exchange resin MIEX(®) (Ixom Watercare Inc.) has been demonstrated to remove substantial amounts of DOM from many source waters, suggesting that MIEX can both reduce DOM content in membrane feed waters and minimize LPM fouling. We tested the effect of MIEX pretreatment on the reduction of short-term LPM fouling potential using feed waters varying in DOM concentration and composition. Four natural and two synthetic waters were studied and a polyvinylidene fluoride (PVDF) hollow-fiber ultrafiltration membrane was used in membrane fouling tests. To evaluate whether MIEX removes the fractions of DOM that cause LPM fouling, the DOM in raw, MIEX-treated, and membrane feed and backwash waters was characterized in terms of DOM concentration and composition. Results showed that: (i) the efficacy of MIEX to reduce LPM fouling varies broadly with source water; (ii) MIEX preferentially removes terrestrial DOM over microbial DOM; (iii) microbial DOM is a more important contributor to LPM fouling than terrestrial DOM, relative to their respective concentrations in source waters; and (iv) the fluorescence intensity of microbial DOM in source waters can be used as a quantitative indicator of the ability of MIEX to reduce their membrane fouling potential. Thus, when ion exchange resin processes are used for DOM removal towards membrane fouling reduction, it is advisable to use a resin that has been designed to effectively remove microbial DOM.

Complete and Partial Photo-oxidation of Dissolved Organic Matter Draining Permafrost Soils.

Photochemical degradation of dissolved organic matter (DOM) to carbon dioxide (CO2) and partially oxidized compounds is an important component of the carbon cycle in the Arctic. Thawing permafrost soils will change the chemical composition of DOM exported to arctic surface waters, but the molecular controls on DOM photodegradation remain poorly understood, making it difficult to predict how inputs of thawing permafrost DOM may alter its photodegradation. To address this knowledge gap, we quantified the susceptibility of DOM draining the shallow organic mat and the deeper permafrost layer of arctic soils to complete and partial photo-oxidation and investigated changes in the chemical composition of each DOM source following sunlight exposure. Permafrost and organic mat DOM had similar lability to photomineralization despite substantial differences in initial chemical composition. Concurrent losses of carboxyl moieties and shifts in chemical composition during photodegradation indicated that photodecarboxylation could account for 40-90% of DOM photomineralized to CO2. Permafrost DOM had a higher susceptibility to partial photo-oxidation compared to organic mat DOM, potentially due to a lower abundance of phenolic moieties with antioxidant properties. These results suggest that photodegradation will likely continue to be an important control on DOM fate in arctic freshwaters as the climate warms and permafrost soils thaw.

Disentangling the interactions between photochemical and bacterial degradation of dissolved organic matter: amino acids play a central role.

Photochemical and bacterial degradation are important pathways to carbon mineralization and can be coupled in dissolved organic matter (DOM) decomposition. However, details of several mechanisms of the coupled photochemical and biological processing of DOM remain too poorly understood to achieve accurate predictions of the impact of these processes on DOM fate and reactivity. The aim of this study was to evaluate how photochemical degradation of amino acids affects bacterial metabolism and whether or not photochemical degradation of DOM competes for amino acids with biological processes. We examined the interactions between photochemical and bacterial degradation dynamics using a mixture of 18 amino acids and examined their dynamics and turnover rates within a larger pool of allochthonous or autochthonous DOM. We observed that photochemical exposure of DOM containing amino acids led to delayed biomass production (even though the final biomass did not differ), most likely due to a need for upregulation of biosynthetic pathways for amino acids that were damaged by photochemically produced reactive oxygen species (ROS). This response was most pronounced in bacterial communities where the abundance of photosensitive amino acids was highest (amended treatments and autochthonous DOM) and least pronounced when the abundance of these amino acids was low (unamended and allochthonous DOM), likely because these bacteria already had these biosynthetic pathways functioning. We observed both a cost and benefit associated with photochemical exposure of DOM. We observed a cost associated with photochemically produced ROS that partially degrade key amino acids and a benefit associated with an increase in the availability of other compounds in the DOM. Bacteria growing on DOM sources that are low in labile amino acids, such as those in terrestrially influenced environments, experience more of the benefits associated with photochemical exposure, whereas bacteria growing in more amino acid-rich environments, such as eutrophic and less terrestrially influenced waters, experience a higher cost due to the increased necessity of salvage pathways for these essential amino acids. Finally, we propose a conceptual model whereby the effects of DOM photochemical degradation on microbial metabolism result from the balance between two mechanisms: One is dependent on the DOM sources, and the other is dependent on the DOM concentration in natural systems.

Carbon cycle. Sunlight controls water column processing of carbon in arctic fresh waters.

Carbon in thawing permafrost soils may have global impacts on climate change; however, the factors that control its processing and fate are poorly understood. The dominant fate of dissolved organic carbon (DOC) released from soils to inland waters is either complete oxidation to CO2 or partial oxidation and river export to oceans. Although both processes are most often attributed to bacterial respiration, we found that photochemical oxidation exceeds rates of respiration and accounts for 70 to 95% of total DOC processed in the water column of arctic lakes and rivers. At the basin scale, photochemical processing of DOC is about one-third of the total CO2 released from surface waters and is thus an important component of the arctic carbon budget.

Insights into the complete and partial photooxidation of black carbon in surface waters.

Increasing wildfire activity in the Alaskan Arctic may result in new sources of black carbon (BC) to arctic watersheds. Black carbon, primarily comprised of condensed aromatics, is one of the most chemically recalcitrant fractions of organic carbon. However, lateral transfer of particulate and dissolved BC from soils to sunlit surface waters is increasingly suggested to result in the photochemical mineralization of BC to CO2. While sunlight can also partially photooxidize aromatic compounds in surface waters, producing compounds with a higher O/C than the parent compound, this degradation pathway has not yet been identified for either particulate or dissolved BC. To address knowledge gaps on the photochemical degradation of particulate and dissolved BC, we quantified the complete and partial photooxidation of particulate and dissolved BC derived from arctic biomass as photochemical CO2 production and O2 consumption relative to dark controls. Concurrently, we investigated shifts in the chemical composition of dissolved BC following exposure to sunlight using UV-visible absorbance, fluorescence spectroscopy, and Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS). The chemical and physical properties of BC produced from charring arctic biomass were similar to BC produced by wildfires in terrestrial ecosystems based on elemental analysis and FT-ICR MS. Based on the concentration of light-absorbing carbon in each fraction, dissolved BC was disproportionately more susceptible to complete and partial photooxidation compared to particulate BC. Upon exposure to sunlight, the predominant fate of dissolved BC was partial photooxidation, while a smaller fraction of dissolved BC was photomineralized to CO2. Shifts in both the optical and mass spectrometry spectra suggested that condensed aromatics likely comprised the fraction of dissolved BC that was completely and partially photooxidized. To further refine the meaning of sunlight as a sink for aquatic BC, the reactivity of partially oxidized photoproducts of BC in the aquatic organic carbon pool must be determined.

Evidence for dissolved organic matter as the primary source and sink of photochemically produced hydroxyl radical in arctic surface waters.

Hydroxyl radical (˙OH) is an indiscriminate oxidant that reacts at near-diffusion-controlled rates with organic carbon. Thus, while ˙OH is expected to be an important oxidant of dissolved organic matter (DOM) and other recalcitrant compounds, the role of ˙OH in the oxidation of these compounds in aquatic ecosystems is not well known due to the poorly constrained sources and sinks of ˙OH, especially in pristine (unpolluted) natural waters. We measured the rates of ˙OH formation and quenching across a range of surface waters in the Arctic varying in concentrations of expected sources and sinks of ˙OH. Photochemical formation of ˙OH was observed in all waters tested, with rates of formation ranging from 2.6 ± 0.6 to 900 ± 100 × 10(-12) M s(-1). Steady-state concentrations ranged from 2 ± 1 to 290 ± 60 × 10(-17) M, and overlapped with previously reported values in surface waters. While iron-mediated photo-Fenton reactions likely contributed to the observed ˙OH production, several lines of evidence suggest that DOM was the primary source and sink of photochemically produced ˙OH in pristine arctic surface waters. DOM from first-order or headwater streams was more efficient in producing ˙OH than what has previously been reported for DOM, and ˙OH formation decreased with increasing residence time of DOM in sunlit surface waters. Despite the ubiquitous formation of ˙OH in arctic surface waters observed in this study, photochemical ˙OH formation was estimated to contribute ≤4% to the observed photo-oxidation of DOM; however, key uncertainties in this estimate must be addressed before ruling out the role of ˙OH in the oxidation of DOM in these waters.

Changes in dissolved organic matter fluorescence and disinfection byproduct formation from UV and subsequent chlorination/chloramination.

Ultraviolet (UV) irradiation is being increasingly used to help drinking water utilities meet finished water quality regulations, but its influence on disinfection byproduct (DBP) precursors and DBP formation is not completely understood. This study investigated the effect of medium pressure (MP) UV combined with chlorination/chloramination on the fluorescent fraction of dissolved organic matter (DOM) isolated from a United States surface water with median total organic carbon content. Parallel factor analysis was used to understand how UV may alter the capacity of DOM to form DBPs of potential human health concern. The production of chloral hydrate and cyanogen chloride from MP UV followed by chlorine or chloramine, respectively, correlated with a decrease in fluorescence intensity of a protein/tryptophan-like component (R(2)=0.79-0.99) and a humic-like component (R(2)=0.91-1.00). This suggests that the UV-induced precursors to these compounds originated from DOM with similar characteristics to these components. The fluorescent DOM components identified in this study are similar to reoccurring components that have been previously identified in a range of raw and treated waters, and this work demonstrates the value of using fluorescence analysis of DOM to understand the relationships between DOM source and DBP formation under a range of treatment conditions.

Dark formation of hydroxyl radical in Arctic soil and surface waters.

Hydroxyl radical (•OH) is a highly reactive and unselective oxidant in atmospheric and aquatic systems. Current understanding limits the role of DOM-produced •OH as an oxidant in carbon cycling mainly to sunlit environments where •OH is produced photochemically, but a recent laboratory study proposed a sunlight-independent pathway in which •OH forms during oxidation of reduced aquatic dissolved organic matter (DOM) and iron. Here we demonstrate this non-photochemical pathway for •OH formation in natural aquatic environments. Across a gradient from dry upland to wet lowland habitats, •OH formation rates increase with increasing concentrations of DOM and reduced iron, with highest •OH formation predicted at oxic-anoxic boundaries in soil and surface waters. Comparison of measured vs expected electron release from reduced moieties suggests that both DOM and iron contribute to •OH formation. At landscape scales, abiotic DOM oxidation by this dark •OH pathway may be as important to carbon cycling as bacterial oxidation of DOM in arctic surface waters.

Determination of specific types and relative levels of QPCR inhibitors in environmental water samples using excitation-emission matrix spectroscopy and PARAFAC.

Assays that utilize PCR offer powerful tools to detect pathogens and other microorganisms in environmental samples. However, PCR inhibitors present in nucleic acid extractions can increase a sample's limit of detection, skew calculated marker concentrations, or cause false-negative results. It would be advantageous to predict which samples contain various types and levels of PCR inhibitors, especially the humic and fulvic acids that are frequently cited as PCR inhibitors in natural water samples. This study investigated the relationships between quantitative PCR (qPCR) inhibition and the humic and fulvic content of dissolved organic matter (DOM), as well as several other measures of DOM quantity and quality, in water samples. QPCR inhibition was also compared to water quality parameters, precipitation levels, and land use adjacent to the sampling location. Results indicate that qPCR inhibition in the tested water samples was correlated to several humic substance-like, DOM components, most notably terrestrially-derived, humic-like DOM and microbially-derived, fulvic-like DOM. No correlation was found between qPCR inhibition and water quality parameters or land use, but a relationship was noted between inhibition and antecedent rainfall. This study suggests that certain fractions of humic substances are responsible for PCR inhibition from temperate, freshwater systems. PARAFAC modeling of excitation-emission matrix spectroscopy provides insight on the components of the DOM pool that impact qPCR success and may be useful in evaluating methods to remove PCR inhibitors present in samples.

Surface exposure to sunlight stimulates CO2 release from permafrost soil carbon in the Arctic.

Recent climate change has increased arctic soil temperatures and thawed large areas of permafrost, allowing for microbial respiration of previously frozen C. Furthermore, soil destabilization from melting ice has caused an increase in thermokarst failures that expose buried C and release dissolved organic C (DOC) to surface waters. Once exposed, the fate of this C is unknown but will depend on its reactivity to sunlight and microbial attack, and the light available at the surface. In this study we manipulated water released from areas of thermokarst activity to show that newly exposed DOC is >40% more susceptible to microbial conversion to CO(2) when exposed to UV light than when kept dark. When integrated over the water column of receiving rivers, this susceptibility translates to the light-stimulated bacterial activity being on average from 11% to 40% of the total areal activity in turbid versus DOC-colored rivers, respectively. The range of DOC lability to microbes seems to depend on prior light exposure, implying that sunlight may act as an amplification factor in the conversion of frozen C stores to C gases in the atmosphere.

Low molecular weight components in an aquatic humic substance as characterized by membrane dialysis and orbitrap mass spectrometry.

Suwannee River fulvic acid (SRFA) was dialyzed through a 100-500 molecular weight cutoff dialysis membrane, and the dialysate and retentate were analyzed by UV-visible absorption and high-resolution Orbitrap mass spectrometry (MS). A significant fraction (36% based on dissolved organic carbon) of SRFA passed through the dialysis membrane. The fraction of SRFA in the dialysate had a different UV-visible absorption spectrum and was enriched in low molecular weight molecules with a more aliphatic composition relative to the initial SRFA solution. Comparison of the SRFA spectra collected by Orbitrap MS and Fourier transform ion cyclotron resonance MS (FT-ICR MS) demonstrated that the mass accuracy of the Orbitrap MS is sufficient for determination of unique molecular formulas of compounds with masses <600 Da in a complex mixture, such as SRFA. The most intense masses detected by Orbitrap MS were found in the 100-200 Da mass range. Many of these low molecular masses corresponded to molecular formulas of previously identified compounds in organic matter, lignin, and plants, and the use of the standard addition method provided an upper concentration estimate of selected target compounds in SRFA. Collectively, these results provide evidence that SRFA contains low molecular weight components that are present individually or in loosely bound assemblies.