Microbial communities change along the 300 km length of the Grand River for extreme high- and low-flow regimes.

The Grand River watershed is the largest catchment in southern Ontario. The river's northern and southern sections are influenced by agriculture, whereas central regions receive wastewater effluent and urban runoff. To characterize in-river microbial communities, as they relate to spatial and environmental factors, we conducted two same-day sampling events along the entire 300 km length of the river, representing contrasting flow seasons (high flow spring melt and low flow end of summer). Through high-throughput sequencing of 16S rRNA genes, we assessed the relationship between river microbiota and spatial and physicochemical variables. Flow season had a greater impact on communities than spatial or diel effects and profiles diverged with distance between sites under both flow conditions, but low-flow profiles exhibited higher beta diversity. High-flow profiles showed greater species richness and increased presence of soil and sediment taxa, which may relate to increased input from terrestrial sources. Total suspended solids, dissolved inorganic carbon, and distance from headwaters significantly explained microbial community variation during the low-flow event, whereas conductivity, sulfate, and nitrite were significant explanatory factors for spring melt. This study establishes a baseline for the Grand River's microbial community, serving as a foundation for modeling the microbiology of anthropogenically impacted freshwater systems affected by lotic processes.

Large Fractionation in Iron Isotopes Implicates Metabolic Pathways for Iron Cycling in Boreal Shield Lakes.

Stable Fe isotopes have only recently been measured in freshwater systems, mainly in meromictic lakes. Here we report the δ56Fe of dissolved, particulate, and sediment Fe in two small dimictic boreal shield headwater lakes: manipulated eutrophic Lake 227, with annual cyanobacterial blooms, and unmanipulated oligotrophic Lake 442. Within the lakes, the range in δ56Fe is large (ca. -0.9 to +1.8‰), spanning more than half the entire range of natural Earth surface samples. Two layers in the water column with distinctive δ56Fe of dissolved (dis) and particulate (spm) Fe were observed, despite differences in trophic states. In the epilimnia of both lakes, a large Δ56Fedis-spm fractionation of 0.4-1‰ between dissolved and particulate Fe was only observed during cyanobacterial blooms in Lake 227, possibly regulated by selective biological uptake of isotopically light Fe by cyanobacteria. In the anoxic layers in both lakes, upward flux from sediments dominates the dissolved Fe pool with an apparent Δ56Fedis-spm fractionation of -2.2 to -0.6‰. Large Δ56Fedis-spm and previously published metagenome sequence data suggest active Fe cycling processes in anoxic layers, such as microaerophilic Fe(II) oxidation or photoferrotrophy, could regulate biogeochemical cycling. Large fractionation of stable Fe isotopes in these lakes provides a potential tool to probe Fe cycling and the acquisition of Fe by cyanobacteria, with relevance for understanding biogeochemical cycling of Earth's early ferruginous oceans.

Early and late cyanobacterial bloomers in a shallow, eutrophic lake.

Cyanobacterial blooms present challenges for water treatment, especially in regions like the Canadian prairies where poor water quality intensifies water treatment issues. Buoyant cyanobacteria that resist sedimentation present a challenge as water treatment operators attempt to balance pre-treatment and toxic disinfection by-products. Here, we used microscopy to identify and describe the succession of cyanobacterial species in Buffalo Pound Lake, a key drinking water supply. We used indicator species analysis to identify temporal grouping structures throughout two sampling seasons from May to October 2018 and 2019. Our findings highlight two key cyanobacterial bloom phases - a mid-summer diazotrophic bloom of Dolichospermum spp. and an autumn Planktothrix agardhii bloom. Dolichospermum crassa and Woronichinia compacta served as indicators of the mid-summer and autumn bloom phases, respectively. Different cyanobacterial metabolites were associated with the distinct bloom phases in both years: toxic microcystins were associated with the mid-summer Dolichospermum bloom and some newly monitored cyanopeptides (anabaenopeptin A and B) with the autumn Planktothrix bloom. Despite forming a significant proportion of the autumn phytoplankton biomass (>60%), the Planktothrix bloom had previously not been detected by sensor or laboratory-derived chlorophyll-a. Our results demonstrate the power of targeted taxonomic identification of key species as a tool for managers of bloom-prone systems. Moreover, we describe an autumn Planktothrix agardhii bloom that has the potential to disrupt water treatment due to its evasion of detection. Our findings highlight the importance of identifying this autumn bloom given the expectation that warmer temperatures and a longer ice-free season will become the norm.

Dissolved oxygen isotope modelling refines metabolic state estimates of stream ecosystems with different land use background.

Dissolved oxygen (DO) is crucial for aerobic life in streams and rivers and mostly depends on photosynthesis (P), ecosystem respiration (R) and atmospheric gas exchange (G). However, climate and land use changes progressively disrupt metabolic balances in natural streams as sensitive reflectors of their catchments. Comprehensive methods for mapping fundamental ecosystem services become increasingly important in a rapidly changing environment. In this work we tested DO and its stable isotope (18O/16O) ratios as novel tools for the status of stream ecosystems. For this purpose, six diel sampling campaigns were performed at three low-order and mid-latitude European streams with different land use patterns. Modelling of diel DO and its stable isotopes combined with land use analyses showed lowest P rates at forested sites, with a minimum of 17.9 mg m-2 h-1. Due to high R rates between 230 and 341 mg m-2 h-1 five out of six study sites showed a general heterotrophic state with P:R:G ratios between 0.1:1.1:1 and 1:1.9:1. Only one site with agricultural and urban influences showed a high P rate of 417 mg m-2 h-1 with a P:R:G ratio of 1.9:1.5:1. Between all sites gross G rates varied between 148 and 298 mg m-2 h-1. In general, metabolic rates depend on the distance of sampling locations to river sources, light availability, nutrient concentrations and possible exchanges with groundwater. The presented modelling approach introduces a new and powerful tool to study effects of land use on stream health. Such approaches should be integrated into future ecological monitoring.

Occurrence of BMAA Isomers in Bloom-Impacted Lakes and Reservoirs of Brazil, Canada, France, Mexico, and the United Kingdom.

The neurotoxic alkaloid ß-N-methyl-amino-l-alanine (BMAA) and related isomers, including N-(2-aminoethyl glycine) (AEG), ß-amino-N-methyl alanine (BAMA), and 2,4-diaminobutyric acid (DAB), have been reported previously in cyanobacterial samples. However, there are conflicting reports regarding their occurrence in surface waters. In this study, we evaluated the impact of amending lake water samples with trichloroacetic acid (0.1 M TCA) on the detection of BMAA isomers, compared with pre-existing protocols. A sensitive instrumental method was enlisted for the survey, with limits of detection in the range of 5−10 ng L−1. Higher detection rates and significantly greater levels (paired Wilcoxon's signed-rank tests, p < 0.001) of BMAA isomers were observed in TCA-amended samples (method B) compared to samples without TCA (method A). The overall range of B/A ratios was 0.67−8.25 for AEG (up to +725%) and 0.69−15.5 for DAB (up to +1450%), with absolute concentration increases in TCA-amended samples of up to +15,000 ng L−1 for AEG and +650 ng L−1 for DAB. We also documented the trends in the occurrence of BMAA isomers for a large breadth of field-collected lakes from Brazil, Canada, France, Mexico, and the United Kingdom. Data gathered during this overarching campaign (overall, n = 390 within 45 lake sampling sites) indicated frequent detections of AEG and DAB isomers, with detection rates of 30% and 43% and maximum levels of 19,000 ng L−1 and 1100 ng L−1, respectively. In contrast, BAMA was found in less than 8% of the water samples, and BMAA was not found in any sample. These results support the analyses of free-living cyanobacteria, wherein BMAA was often reported at concentrations of 2−4 orders of magnitude lower than AEG and DAB. Seasonal measurements conducted at two bloom-impacted lakes indicated limited correlations of BMAA isomers with total microcystins or chlorophyll-a, which deserves further investigation.

Quantifying arsenic post-depositional mobility in lake sediments impacted by gold ore roasting in sub-arctic Canada using inverse diagenetic modelling.

Lake sediments are widely used as environmental archives to reconstruct past changes in contaminants deposition, provided that they remain immobile after deposition. Arsenic (As) is a redox-sensitive element that may be redistributed in the sediments during early diagenesis, for instance along with iron and manganese, and thus depth profiles of As might not provide a reliable, unaltered record of past deposition. Here, we use inverse diagenetic modelling to calculate fluxes of As across the sediment-water interface and interpret As sedimentary records in eight lakes along a 80 km transect from the Giant and Con mines, Northwest Territories, Canada. The sediment cores were dated using 210Pb methods and analyzed for solid-phase and porewater As, Fe, Mn and organic C concentrations. We reconstructed the history of As deposition by correcting for the varying mobility patterns and calculated contemporary As deposition fluxes. Correction for diagenesis was substantial for three of the eight lakes, suggesting that lakes with lower sedimentation rates, which allows longer residence of As within the reactive zones defined by the model, enhance the influence of diagenesis. Results show that solid phase As peaks coincides with the period of high emissions from past gold ore roasting activities. Results also show that sediments sustained present-day As fluxes to the water column of study lakes within 50 km of the mines, while sediment in study lakes further than 50 km acted as As sinks instead.

Dairy manure acidification reduces CH4 emissions over short and long-term.

Acidification with sulphuric acid and cleaning residual manure in tanks are promising practices for reducing methane (CH4), which is a potent greenhouse gas. To date, no data are available on CH4 reductions from acidifying only residual manure (rather than all manure). Moreover, long-term effects of manure acidification (i.e. inoculating ability of previously acidified residual manure in the subsequent storages) are not known. To address these gaps, fresh manure (FM; 150 mL) combined with treated or untreated inoculum (30 mL) were anaerobically incubated at 17°C, 20°C, and 23°C for 116 d. Acidified treatments, regardless of location of acid addition, reduced CH4 production by 81% at 17°C, 78% at 20°C, and 19% at 23°C compared to the control (untreated FM and untreated inoculum). To test long-term acidification effects, FM was inoculated with manure that had been acidified 6-months prior. This created comparable CH4 production to FM with no inoculum and reduced CH4 production by 99% at 17°C and 20°C, and 49% at 23°C compared to the control. Results indicate that residual slurries of acidified manure become poor inoculants in subsequent storage, hence manure acidification has a long-term treatment effect in reducing CH4 production. This could reduce how often acidification is needed in dairy manure tanks and also increasing its cost-effectiveness for farmers.

Greenhouse Gas Mitigation through Dairy Manure Acidification.

Liquid dairy manure storages are sources of methane (CH), nitrous oxide (NO), and ammonia (NH) emissions. Both CH and NO are greenhouse gases (GHGs), whereas NH is an indirect source of NO emissions. Manure acidification is a strategy used to reduce NH emissions from swine manure; however, limited research has expanded this strategy to reducing CH and NO emissions by acidifying dairy manure. This study compared control dairy manure (pH 7.4) with two treatments of acidified manure using 70% sulfuric acid (HSO). These included a medium pH treatment (pH 6.5, 1.4 mL acid L manure) and a low pH treatment (pH 6, 2.4 mL acid L manure). Emissions were measured using replicated mesoscale manure tanks (6.6 m) enclosed by large steady state chambers. Both CH and NO were continuously measured (June-December 2017) using tunable diode laser trace gas analyzers. Ammonia emissions were measured three times weekly for 24 h using acid traps. On a CO equivalent basis, the medium pH treatment reduced total GHG emissions by 85%, whereas the low pH treatment reduced emissions by 88%, relative to untreated (control) manure. Total CH emissions were reduced by 87 and 89% from medium and low pH tanks, respectively. Ammonia emissions were reduced by 41 and 53% from medium and low pH tanks, respectively. Additional research is necessary to make acidification an accessible option for farmers by optimizing acid dosage. More research is need to describe the manure buffering capacity and emission reductions and ultimately find the best approaches for treating farm-scale liquid dairy manure tanks.

Multiple sources and sinks of dissolved inorganic carbon across Swedish streams, refocusing the lens of stable C isotopes.

It is well established that stream dissolved inorganic carbon (DIC) fluxes play a central role in the global C cycle, yet the sources of stream DIC remain to a large extent unresolved. Here, we explore large-scale patterns in δ13C-DIC from streams across Sweden to separate and further quantify the sources and sinks of stream DIC. We found that stream DIC is governed by a variety of sources and sinks including biogenic and geogenic sources, CO2 evasion, as well as in-stream processes. Although soil respiration was the main source of DIC across all streams, a geogenic DIC influence was identified in the northernmost region. All streams were affected by various degrees of atmospheric CO2 evasion, but residual variance in δ13C-DIC also indicated a significant influence of in-stream metabolism and anaerobic processes. Due to those multiple sources and sinks, we emphasize that simply quantifying aquatic DIC fluxes will not be sufficient to characterise their role in the global C cycle.

From the ground up: global nitrous oxide sources are constrained by stable isotope values.

Rising concentrations of nitrous oxide (N2O) in the atmosphere are causing widespread concern because this trace gas plays a key role in the destruction of stratospheric ozone and it is a strong greenhouse gas. The successful mitigation of N2O emissions requires a solid understanding of the relative importance of all N2O sources and sinks. Stable isotope ratio measurements (δ15N-N2O and δ18O-N2O), including the intramolecular distribution of 15N (site preference), are one way to track different sources if they are isotopically distinct. 'Top-down' isotope mass-balance studies have had limited success balancing the global N2O budget thus far because the isotopic signatures of soil, freshwater, and marine sources are poorly constrained and a comprehensive analysis of global N2O stable isotope measurements has not been done. Here we used a robust analysis of all available in situ measurements to define key global N2O sources. We showed that the marine source is isotopically distinct from soil and freshwater N2O (the continental source). Further, the global average source (sum of all natural and anthropogenic sources) is largely controlled by soils and freshwaters. These findings substantiate past modelling studies that relied on several assumptions about the global N2O cycle. Finally, a two-box-model and a Bayesian isotope mixing model revealed marine and continental N2O sources have relative contributions of 24-26% and 74-76% to the total, respectively. Further, the Bayesian modeling exercise indicated the N2O flux from freshwaters may be much larger than currently thought.

Large carbon dioxide fluxes from headwater boreal and sub-boreal streams.

Half of the world's forest is in boreal and sub-boreal ecozones, containing large carbon stores and fluxes. Carbon lost from headwater streams in these forests is underestimated. We apply a simple stable carbon isotope idea for quantifying the CO2 loss from these small streams; it is based only on in-stream samples and integrates over a significant distance upstream. We demonstrate that conventional methods of determining CO2 loss from streams necessarily underestimate the CO2 loss with results from two catchments. Dissolved carbon export from headwater catchments is similar to CO2 loss from stream surfaces. Most of the CO2 originating in high CO2 groundwaters has been lost before typical in-stream sampling occurs. In the Harp Lake catchment in Canada, headwater streams account for 10% of catchment net CO2 uptake. In the Krycklan catchment in Sweden, this more than doubles the CO2 loss from the catchment. Thus, even when corrected for aquatic CO2 loss measured by conventional methods, boreal and sub-boreal forest carbon budgets currently overestimate carbon sequestration on the landscape.

Proper interpretation of dissolved nitrous oxide isotopes, production pathways, and emissions requires a modelling approach.

Stable isotopes ([Formula: see text]15N and [Formula: see text]18O) of the greenhouse gas N2O provide information about the sources and processes leading to N2O production and emission from aquatic ecosystems to the atmosphere. In turn, this describes the fate of nitrogen in the aquatic environment since N2O is an obligate intermediate of denitrification and can be a by-product of nitrification. However, due to exchange with the atmosphere, the [Formula: see text] values at typical concentrations in aquatic ecosystems differ significantly from both the source of N2O and the N2O emitted to the atmosphere. A dynamic model, SIDNO, was developed to explore the relationship between the isotopic ratios of N2O, N2O source, and the emitted N2O. If the N2O production rate or isotopic ratios vary, then the N2O concentration and isotopic ratios may vary or be constant, not necessarily concomitantly, depending on the synchronicity of production rate and source isotopic ratios. Thus prima facie interpretation of patterns in dissolved N2O concentrations and isotopic ratios is difficult. The dynamic model may be used to correctly interpret diel field data and allows for the estimation of the gas exchange coefficient, N2O production rate, and the production-weighted [Formula: see text] values of the N2O source in aquatic ecosystems. Combining field data with these modelling efforts allows this critical piece of nitrogen cycling and N2O flux to the atmosphere to be assessed.

Nonlinear response of riverine N2O fluxes to oxygen and temperature.

One-quarter of anthropogenically produced nitrous oxide (N2O) comes from rivers and estuaries. Countries reporting N2O fluxes from aquatic surfaces under the United Nations Framework Convention on Climate Change typically estimate anthropogenic inorganic nitrogen loading and assume a fraction becomes N2O. However, several studies have not confirmed a linear relationship between dissolved nitrate (NO3-) and river N2O fluxes. We apply recursive partitioning analysis to examine the relationships between N2O flux and NO3-, dissolved oxygen (DO), temperature, land use and surficial geology in the Grand River, Canada, a seventh-order river in an agricultural catchment with substantial urban population. Results suggest that N2O flux is high when hypoxia exists. Temperature, not NO3-, was the primary correlate of N2O flux when hypoxia does not occur suggesting NO3- is not limiting N2O production and further increases in NO3- may not lead to comparable increases in N2O flux. This work indicates that a linear relationship between NO3- and N2O is unlikely to exist in most agricultural and urban impacted river systems. Most N2O is produced during hypoxia so quantifying the extent of hypoxia is a necessary first step to quantifying N2O fluxes in lotic systems. Predicted increases in riverine hypoxia via eutrophication and increased temperature due to climate change may drive nonlinear increases in N2O production.

Stable oxygen isotope ratios of nitrate produced from nitrification: (18)O-labeled water incubations of agricultural and temperate forest soils.

In many nitrate (NO(3)(-)) source partitioning studies, the delta(18)O value for NO(3)(-) produced from nitrification is often assumed to reflect the isotopic compositions of environmental water (H(2)O) and molecular oxygen (O(2)) in a 2:1 ratio. Most studies that have measured or observed this microbial endmember have found that the delta(18)O-NO(3)(-) was more positive (up to +15 per thousand higher) than the assumed value. Current understanding of the mechanism(s) responsible for this discrepancy is limited. Incubations of one temperate forest soil (organic) and two agricultural soils (mineral) were conducted with (18)O-labeled H(2)O to apportion the sources of oxygen in NO(3)(-) generated from nitrification. The NO(3)(-) produced in all soils had delta(18)O values that could not be explained by a simple endmember mixing ratio of 2:1. A more comprehensive model describing the formation of microbial NO(3)(-) was developed, which accounts for oxygen exchange between H(2)O and NO(2)(-), and includes terms for kinetic and equilibrium isotope effects. Oxygen isotope exchange (i.e., the fraction of NO(3)(-)-oxygen that originates from the abiotic exchange of H(2)O and NO(2)(-)) varied widely between the temperate forest soil (0.37) and the two agricultural soils (0.52 and 0.88). At present, the microbial endmember for nitrification cannot be successfully predicted.

Aquatic metabolism and ecosystem health assessment using dissolved O2 stable isotope diel curves.

Dissolved O2 concentration and delta18O-O2 diel curves can be combined to assess aquatic photosynthesis, respiration, and metabolic balance, and to disentangle some of the confounding factors associated with interpretation of traditional O2 concentration curves. A dynamic model is used to illustrate how six key environmental and biological parameters interact to affect diel O2 saturation and delta18O-O2 curves, thereby providing a fundamental framework for the use of delta18O-O2 in ecosystem productivity studies. delta18O-O2 provides information unavailable from concentration alone because delta18O-O2 and saturation curves are not symmetrical and can be used to constrain gas exchange and isotopic fractionation by eliminating many common assumptions. Changes in key parameters affect diel O2 saturation and delta18O-O2 curves as follows: (1) an increase in primary production and respiration rates increases the diel range of O2 saturation and delta18O-O2 and decreases the mean delta18O-O2 value; (2) a decrease in the primary production to respiration ratio (P:R) decreases the level of O2 saturation and increases the delta18O-O2 values; (3) an increase in the gas exchange rate decreases the diel range of O2 saturation and delta18O-O2 values and moves the mean O2 saturation and delta18O-O2 values toward atmospheric equilibrium; (4) a decrease in strength of the respiratory isotopic fractionation (alphaR closer to 1) has no effect on O2 saturation and decreases the delta18O-O2 values; (5) an increase in the delta18O of water has no effect on O2 saturation and increases the minimum (daytime) delta18O-O2 value; and (6) an increase in temperature reduces O2 solubility and thus increases the diel range of O2 saturation and delta18O-O2 values. Understanding the interplay between these key parameters makes it easier to decipher the controls on O2 and delta18O-O2, compare aquatic ecosystems, and make quantitative estimates of ecosystem metabolism. The photosynthesis to respiration to gas exchange ratio (P:R:G) is better than the P:R ratio at describing and assessing the vulnerability of aquatic ecosystems under various environmental stressors by providing better constrained estimates of ecosystem metabolism and gas exchange.

Dynamics of dissolved oxygen isotopic ratios: a transient model to quantify primary production, community respiration, and air-water exchange in aquatic ecosystems.

Dissolved O(2) is an important aquatic ecosystem health indicator. Metabolic and gas exchange (G) rates, which control O(2) concentration, are affected by nutrient loading and other environmental factors. Traditionally, aquatic metabolism has been reported as primary production:community respiration (P:R) ratios using diel measurements and interpretations of dissolved O(2) and/or CO(2) concentrations, and recently using stable isotopes (delta(18)O, Delta(17)O) and steady state assumptions. Aquatic ecosystems, such as rivers and ponds, are not at steady state and exhibit diel changes, so steady state approaches are often inappropriate. A dynamic O(2) stable isotope model (photosynthesis-respiration-gas exchange; PoRGy) is presented here, requiring a minimum of parameters to quantify daily averaged P, R, and G rates under transient field conditions. Unlike steady state approaches, PoRGy can address scenarios with 100% O(2) saturation but with delta(18)O-O(2) values that are not at air equilibrium. PoRGy successfully accounts for isotopic G when applied to an oxygen isotope equilibration laboratory experiment. PoRGy model results closely matched the diel O(2) and delta(18)O-O(2) data from three field sites with different P:R:G ratios and various P, R and G rates. PoRGy provides a new research tool to assess ecosystem health and to pose environmental impact-driven questions. Using daily averaged rates was successful and thus they can be used to compare ecosystems across seasons and landscapes.