Proteome changes in larval zebrafish (Danio rerio) and fathead minnow (Pimephales promelas) exposed to (±) anatoxin-a.

Anatoxin-a and its analogues are potent neurotoxins produced by several genera of cyanobacteria. Due in part to its high toxicity and potential presence in drinking water, these toxins pose threats to public health, companion animals and the environment. It primarily exerts toxicity as a cholinergic agonist, with high affinity at neuromuscular junctions, but molecular mechanisms by which it elicits toxicological responses are not fully understood. To advance understanding of this cyanobacteria, proteomic characterization (DIA shotgun proteomics) of two common fish models (zebrafish and fathead minnow) was performed following  (±) anatoxin-a exposure. Specifically, proteome changes were identified and quantified in larval fish exposed for 96 h (0.01-3 mg/L (±) anatoxin-a and caffeine (a methodological positive control) with environmentally relevant treatment levels examined based on environmental exposure distributions of surface water data. Proteomic concentration - response relationships revealed 48 and 29 proteins with concentration - response relationships curves for zebrafish and fathead minnow, respectively. In contrast, the highest number of differentially expressed proteins (DEPs) varied between zebrafish (n = 145) and fathead minnow (n = 300), with only fatheads displaying DEPs at all treatment levels. For both species, genes associated with reproduction were significantly downregulated, with pathways analysis that broadly clustered genes into groups associated with DNA repair mechanisms. Importantly, significant differences in proteome response between the species was also observed, consistent with prior observations of differences in response using both behavioral assays and gene expression, adding further support to model specific differences in organismal sensitivity and/or response. When DEPs were read across from humans to zebrafish, disease ontology enrichment identified diseases associated with cognition and muscle weakness consistent with the prior literature. Our observations highlight limited knowledge of how (±) anatoxin-a, a commonly used synthetic racemate surrogate, elicits responses at a molecular level and advances its toxicological understanding.

The effects of salinity and N:P on N-rich toxins by both an N-fixing and non-N-fixing cyanobacteria.

Freshwater ecosystems are experiencing increased salinization. Adaptive management of harmful algal blooms (HABs) contribute to eutrophication/salinization interactions through the hydrologic transport of blooms to coastal environments. We examined how nutrients and salinity interact to affect growth, elemental composition, and cyanotoxin production/release in two common HAB genera. Microcystis aeruginosa (non-nitrogen (N)-fixer and microcystin-LR producer; MC-LR) and Aphanizomenon flos-aquae (N-fixer and cylindrospermopsin producer; CYN) were grown in N:phosphorus (N:P) 4 and 50 (by atom) for 21 and 33 days, respectively, then dosed with a salinity gradient (0 - 10.5 g L-1). Both total MC-LR and CYN were correlated with particulate N. We found Microcystis MC-LR production and release was affected by salinity only in the N:P 50 treatment. However, Aphanizomenon CYN production and release was affected by salinity regardless of N availability. Our results highlight how cyanotoxin production and release across the freshwater - marine continuum are controlled by eco-physiological differences between N-acquisition traits.

Diazotrophy modulates cyanobacteria stoichiometry through functional traits that determine bloom magnitude and toxin production.

Harmful cyanobacterial blooms are an increasing threat to water quality. The interactions between two eco-physiological functional traits of cyanobacteria, diazotrophy (nitrogen (N)-fixation) and N-rich cyanotoxin synthesis, have never been examined in a stoichiometric explicit manner. We explored how a gradient of resource N:phosphorus (P) affects the biomass, N, P stoichiometry, light-harvesting pigments, and cylindrospermopsin production in a N-fixing cyanobacterium, Aphanizomenon. Low N:P Aphanizomenon cultures produced the same biomass as populations grown in high N:P cultures. The biomass accumulation determined by carbon, indicated low N:P Aphanizomenon cultures did not have a N-fixation growth tradeoff, in contrast to some other diazotrophs that maintain stoichiometric N homeostasis at the expense of growth. However, N-fixing Aphanizomenon populations produced less particulate cylindrospermopsin and had undetectable dissolved cylindrospermopsin compared to non-N-fixing populations. The pattern of low to high cyanotoxin cell quotas across an N:P gradient in the diazotrophic cylindrospermopsin producer is similar to the cyanotoxin cell quota response in non-diazotrophic cyanobacteria. We suggest that diazotrophic cyanobacteria may be characterized into two broad functional groups, the N-storage-strategists and the growth-strategists, which use N-fixation differently and may determine patterns of bloom magnitude and toxin production in nature.

Comment on "Models predict planned phosphorus load reduction will make Lake Erie more toxic".

Hellweger et al. (Reports, 27 May 2022, pp. 1001) predict that phosphorus limitation will increase concentrations of cyanobacterial toxins in lakes. However, several molecular, physiological, and ecological mechanisms assumed in their models are poorly supported or contradicted by other studies. We conclude that their take-home message that phosphorus load reduction will make Lake Erie more toxic is seriously flawed.

Incorporating parameter variability into Monod models of nutrient-limited growth of non-diazotrophic and diazotrophic cyanobacteria.

Models are widely used tools in aquatic science to understand the mechanism of phytoplankton growth and anticipate the occurrence of harmful algal blooms. However, model parameterization remains challenging and issues that may introduce prediction uncertainty exist. Many models use the Monod equation to predict cyanobacteria growth rate based on ambient nutrient concentrations. The half-saturation concentrations in the Monod equation varies greatly among different studies and depends on environmental conditions. In this study, we estimated the growth rate due to nutrient limitations for two cyanobacteria species (Microcystis aeruginosa and Dolichospermum flos-aquae) using a modified Monod model which allows the half-saturation concentration to vary according to initial nitrogen (N) conditions. The model is calibrated against observations from laboratory experiment where cyanobacteria growth and ambient nutrient concentrations were measured simultaneously, which is rarely done in the literature. Our results show this modified model produce better predictions on growth rate and biomass, indicating many commonly used mechanistic models may need improvement regarding phytoplankton growth representation. Furthermore, our study quantifies the flexibility in cyanobacteria growth parameter across a wide range of environmental N in eutrophic lakes thus provides important information for large-scale modelling applications.

Dynamic Phycobilin Pigment Variations in Diazotrophic and Non-diazotrophic Cyanobacteria Batch Cultures Under Different Initial Nitrogen Concentrations.

Increased anthropogenic nutrient loading has led to eutrophication of aquatic ecosystems, which is the major cause of harmful cyanobacteria blooms. Element stoichiometry of cyanobacteria bloom is subject to nutrient availabilities and may significantly contribute to primary production and biogeochemical cycling. Phycobilisome is the antenna of the photosynthetic pigment apparatus in cyanobacteria, which contains phycobilin pigments (PBPs) and linker proteins. This nitrogen (N)-rich protein complex has the potential to support growth as a N-storage site and may play a major role in the variability of cyanobacteria N stoichiometry. However, the regulation of PBPs during bloom formation remains unclear. We investigated the temporal variation of N allocation into PBPs and element stoichiometry for two ubiquitous cyanobacteria species, Microcystis aeruginosa and Dolichospermum flos-aquae, in a batch culture experiment with different initial N availabilities. Our results indicated that the N allocation into PBPs is species-dependent and tightly regulated by the availability of nutrients fueling population expansion. During the batch culture experiment, different nutrient uptake rates led to distinct stoichiometric imbalances of N and phosphorus (P), which substantially altered cyanobacteria C: N and C: P stoichiometry. Microcystis invested cellular N into PBPs and exhibited greater flexibility in C: N and C: P stoichiometry than D. flos-aquae. The dynamics of such N-rich macromolecules may help explain the N stoichiometry variation during a bloom and the interspecific difference between M. aeruginosa and D. flos-aquae. Our study provides a quantitative understanding of the elemental stoichiometry and the regulation of PBPs for non-diazotrophic and diazotrophic cyanobacteria blooms.

Nitrogen fixation: a poorly understood process along the freshwater-marine continuum.

Although N2 fixation is a major component of the global N cycle and has been extensively studied in open-ocean and terrestrial ecosystems, rates and ecological dynamics remain virtually unknown for the inland and coastal aquatic ecosystems (lakes, wetlands, rivers, streams, estuaries) that connect terrestrial and marine biomes. This is due to the diversity of these habitats, as well as the traditional paradigm that N2 fixation rates were low to nonexistent, and therefore not important, in these ecosystems. We identify three major research themes to advance understanding of aquatic N2 fixation: 1) the biological diversity of diazotrophs and variability of N2 fixation rates, 2) the ecological stoichiometry of N2 fixation, and 3) the upscaling of N2 fixation rates from genes to ecosystems. Coordinating research across these areas will advance limnology and oceanography by fully integrating N2 fixation into ecological dynamics of aquatic ecosystems from local to global scales.

Differential influences of (±) anatoxin-a on photolocomotor behavior and gene transcription in larval zebrafish and fathead minnows.

BACKGROUND: Though anatoxin-a (antx-a) is a globally important cyanobacterial neurotoxin in inland waters, information on sublethal toxicological responses of aquatic organisms is limited. We examined influences of (±) antx-a (11-3490 µg/L) on photolocomotor behavioral responses and gene transcription associated with neurotoxicity, oxidative stress and hepatotoxicity, in two of the most common alternative vertebrate and fish models, Danio rerio (zebrafish) and Pimephales promelas (fathead minnow). We selected environmentally relevant treatment levels from probabilistic exposure distributions, employed standardized experimental designs, and analytically verified treatment levels using isotope-dilution liquid chromatography tandem mass spectrometry. Caffeine was examined as a positive control. RESULTS: Caffeine influences on fish behavior responses were similar to previous studies. Following exposure to (±) antx-a, no significant photolocomotor effects were observed during light and dark transitions for either species. Though zebrafish behavioral responses profiles were not significantly affected by (±) antx-a at the environmentally relevant treatment levels examined, fathead minnow stimulatory behavior was significantly reduced in the 145-1960 µg/L treatment levels. In addition, no significant changes in transcription of target genes were observed in zebrafish; however, elavl3 and sod1 were upregulated and gst and cyp3a126 were significantly downregulated in fathead minnows. CONCLUSION: We observed differential influences of (±) antx-a on swimming behavior and gene transcription in two of the most common larval fish models employed for prospective and retrospective assessment of environmental contaminants and water quality conditions. Sublethal responses of fathead minnows were consistently more sensitive than zebrafish to this neurotoxin at the environmentally relevant concentrations examined. Future studies are needed to understand such interspecies differences, the enantioselective toxicity of this compound, molecular initiation events within adverse outcome pathways, and subsequent individual and population risks for this emerging water quality threat.

Nitrogen form, concentration, and micronutrient availability affect microcystin production in cyanobacterial blooms.

Harmful algal blooms (HABs) are increasing in magnitude, frequency, and duration caused by anthropogenic factors such as eutrophication and altered climatic regimes. While the concentrations and ratios of nitrogen (N) and phosphorus are correlated with bloom biomass and cyanotoxin production, there is less known about how N forms and micronutrients (MN) interact to regulate HABs and cyanotoxin production. Here, we used two separate approaches to examine how N and MN supply affects cyanobacteria biomass and cyanotoxin production. First, we used a Microcystis laboratory culture to examine how N and MN concentration and N form affected the biomass, particulate N, and microcystin-LR concentration and cell quotas. Then, we monitored the N, iron, molybdenum, and total microcystin concentrations from a hypereutrophic reservoir. From this hypereutrophic reservoir, we performed a community HAB bioassay to examine how N and MN addition affected the biomass, particulate N, and microcystin concentration. Microcystis laboratory cultures grown in high urea and MN conditions produced more biomass, particulate N, and had similar C:N stoichiometry, but lower microcystin-LR concentrations and cell quotas when compared to high nitrate and MN conditions. Our community HAB bioassay revealed no interactions between N concentration and MN addition caused by non-limiting MN background concentrations. Biomass, particulate N, and microcystin concentration increased with N addition. The community HAB amended with MN resulted in greater microcystin-LA concentration compared to non-MN amended community HABs. Our results highlight the complexity of how abiotic variables control biomass and cyanotoxin production in both laboratory cultures of Microcystis and community HABs.

Biological stoichiometry and growth dynamics of a diazotrophic cyanobacteria in nitrogen sufficient and deficient conditions.

The role of nitrogen (N) fixation in determining the frequency, magnitude, and extent of harmful algal blooms (HABs) has not been well studied. Dolichospermum is a common HAB species that is diazotrophic (capable of N fixation) and thus growth is often considered never to be limited by low combined N sources. However, N fixation is energetically expensive and its cost during bloom formation has not been quantified. Additionally, it is unknown how acclimation to differing nutrient ratios affects growth and cellular carbon (C):N stoichiometry. Here, we test the hypotheses that diazotrophic cyanobacteria are homeostatic for N because of their ability to fix atmospheric N2 and that previous acclimation to low N environments will result in more fixed N and lower C:N stoichiometry. Briefly, cultures that varied in resource N:phosphorus (P) ranging from 0.01 to 100 (atom), were seeded with Dolichospermum which were previously acclimated to low and high N:P conditions and then sampled temporally for growth and C:N stoichiometry. We found that Dolichospermum was not homeostatic for N and displayed classic signs of N limitation and elevated C:N stoichiometry, highlighting the necessary growth trade-off within cells when expending energy to fix N. Acclimation to N limited conditions caused differences in both C:N and fixed N at various time points in the experiment. These results highlight the importance of environmentally available N to a diazotrophic bloom, as well as how previous growth conditions can influence population growth during blooms experiencing variable N:P.

Stoichiometric imbalances complicate prediction of phytoplankton biomass in U.S. lakes: implications for nutrient criteria.

Using National Lakes Assessment data, we evaluated the influence of total N (TN), total P (TP), and other variables on lake chlorophyll-a concentrations. With simple linear regressions, high TN/TP samples biased predictions based on TN, and low TN/TP samples biased predictions based on TP. The bias problem was corrected, and correlation was improved, by splitting the dataset at the TN/TP ratio we estimated to be indicative of a balanced supply and developing separate regressions that predict chlorophyll-a based on TP, TN, dissolved inorganic N (DIN), dissolved organic carbon (DOC), non-algal light attenuation, depth, area, latitude, elevation, and conductivity. Both nutrients were excellent predictors, and non-algal light attenuation was the next most influential predictor. The regression analysis suggested that a potential for P only limitation (high TN/TP, 17% of samples) or N only limitation (low TN/TP, 14% of samples) can be inferred at the extremes of the TN/TP range. However, 69% of samples had an intermediate TN/TP ratio where it is difficult to infer anything about potential nutrient limitations (biomass could be N limited, P limited, N and P co-limited, or not limited by nutrients at all). Our results show that when developing phytoplankton response relationships using cross-lake datasets that span a wide range of trophic states, it is important to consider whether and how biomass is influenced by confounding factors - such as differences in the relative supply of N and P - so that biomass is not underestimated or overestimated, and nutrient criteria are not under-protective or over-protective.

Stream algal biomass response to experimental phosphorus and nitrogen gradients: A case for dual nutrient management in agricultural watersheds.

Watershed managers generally focus on P reduction strategies to combat freshwater eutrophication despite evidence that N co-limits primary production. Our objective was to test the role of P in limiting stream periphyton biomass within the Buffalo River watershed in Arkansas by conducting a 31-d streamside mesocosm experiment. To represent potentially different starting states, cobbles were transplanted from two different tributary streams and initially exposed to a range of P (0, 0.012, 0.025, 0.05, 0.1, and 0.2 mg L-1 P) to assess benthic ash-free dry mass (AFDM) and chlorophyll-a (chl a) and responses during a P only enrichment period. Later, the experiment was continued under a N/P (10:1 molar ratio) enrichment gradient to examine co-limitation. Mean AFDM was higher on Day 31 of the N+P enrichment compared with Day 17 of the P-only enrichment (p < .001). Overall differences in AFDM and chl a were observed between cobbles from different stream sites. Phosphorus enrichment stimulated benthic chl a biomass, but enrichment effects were greater when streams were enriched with N+P (p < .001). Chlorophyll-a increased (4.4-57.9 mg m-2 ) with increasing P concentrations (p < .001) after P enrichment but was threefold greater after N+P enrichment, increasing from 13.3 to 171.1 mg m-2 across the enrichment gradient. Results support the need to consider both N and P limitation in freshwater systems and demonstrate that potential increases in nutrient concentrations may influence accumulation of algae on cobble substrates from the Buffalo River watershed.

Global scanning of cylindrospermopsin: Critical review and analysis of aquatic occurrence, bioaccumulation, toxicity and health hazards.

Cylindrospermopsin (CYN), a cyanotoxin produced by harmful algal blooms, has been reported worldwide; however, there remains limited understanding of its potential risks to surface water quality. In the present study, we critically reviewed available literature regarding the global occurrence, bioaccumulation, and toxicity of CYN in aquatic systems with a particular focus on freshwater. We subsequently developed environmental exposure distributions (EEDs) for CYN in surface waters and performed probabilistic environmental hazard assessments (PEHAs) using guideline values (GVs). PEHAs were performed by geographic region, type of aquatic system, and matrix. CYN occurrence was prevalent in North America, Europe, and Asia/Pacific, with lakes being the most common system. Many global whole water EEDs exceeded guideline values (GV) previously developed for drinking water (e.g., 0.5 µg L-1) and recreational water (e.g., 1 µg L-1). GV exceedances were higher in the Asia/Pacific region, and in rivers and reservoirs. Rivers in the Asia/Pacific region exceeded the lowest drinking water GV 73.2% of the time. However, lack of standardized protocols used for analyses was alarming, which warrants improvement in future studies. In addition, bioaccumulation of CYN has been reported in mollusks, crustaceans, and fish, but such exposure information remains limited. Though several publications have reported aquatic toxicity of CYN, there is limited chronic aquatic toxicity data, especially for higher trophic level organisms. Most aquatic toxicity studies have not employed standardized experimental designs, failed to analytically verify treatment levels, and did not report purity of CYN used for experiments; therefore, existing data are insufficient to derive water quality guidelines. Considering such elevated exceedances of CYN in global surface waters and limited aquatic bioaccumulation and toxicity data, further aquatic monitoring, environmental fate and mechanistic toxicology studies are warranted to robustly assess and manage water quality risks to public health and the environment.

Biological Stoichiometry Regulates Toxin Production in Microcystis aeruginosa (UTEX 2385).

Harmful algal blooms (HABs) are increasing in magnitude, frequency, and duration globally. Even though a limited number of phytoplankton species can be toxic, they are becoming one of the greatest water quality threats to public health and ecosystems due to their intrinsic toxicity to humans and the numerous interacting factors that undermine HAB forecasting. Here, we show that the carbon:nitrogen:phosphorus (C:N:P) stoichiometry of a common toxic phytoplankton species, Microcystis, regulates toxin quotas during blooms through a tradeoff between primary and secondary metabolism. Populations with optimal C:N (< 8) and C:P (< 200) cellular stoichiometry consistently produced more toxins than populations exhibiting stoichiometric plasticity. Phosphorus availability in water exerted a strong control on population biomass and C:P stoichiometry, but N availability exerted a stronger control on toxin quotas by regulating population biomass and C:N:P stoichiometry. Microcystin-LR, like many phytoplankton toxins, is an N-rich secondary metabolite with a C:N stoichiometry that is similar to the optimal growth stoichiometry of Microcystis. Thus, N availability relative to P and light provides a dual regulatory mechanism that controls both biomass production and cellular toxin synthesis. Overall, our results provide a quantitative framework for improving forecasting of toxin production during HABs and compelling support for water quality management that limit both N and P inputs from anthropogenic sources.

Interspecific homeostatic regulation and growth across aquatic invertebrate detritivores: a test of ecological stoichiometry theory.

Across resource quality gradients, primary consumers must regulate homeostasis and release of nutrients to optimize growth and fitness. Based primarily on internal body composition, the ecological stoichiometry theory (EST) offers a framework to generalize interspecific patterns of these responses, yet the predictions and underlying assumptions of EST remain poorly tested across many species. We used controlled laboratory feeding experiments to measure homeostasis, nutrient release, and growth across seven field-collected aquatic invertebrate detritivore taxa fed wide resource carbon:nitrogen (C:N) and carbon:phosphorus (C:P) gradients. We found that most invertebrates exhibited strict stoichiometric homeostasis (average 1/H = - 0.018 and 0.026 for C:N and C:P, respectively), supporting assumptions of EST. However, the stoichiometry of new tissue production during growth intervals (growth stoichiometry) deviated - 30 to + 54% and - 145 to + 74% from initial body C:N and C:P, respectively, and across species, growth stoichiometry was not correlated with initial body stoichiometry. Notably, smaller non- and hemimetabolous invertebrates exhibited low, decreasing growth C:N and C:P, whereas larger holometabolous invertebrates exhibited high, often increasing growth C:N and C:P. Despite predictions of EST, interspecific sensitivity of egestion stoichiometry and growth rates to the resource gradient were weakly related to internal body composition across species. While the sensitivity of these patterns differed across taxa, such differences carried a weak phylogenetic signal and were not well predicted by EST. Our findings suggest that traits beyond internal body composition, such as feeding behavior, selective assimilation, and ontogeny, are needed to generalize interspecific patterns in consumer growth and nutrient release across resource quality gradients.

Substituting values for censored data from Texas, USA, reservoirs inflated and obscured trends in analyses commonly used for water quality target development.

We calculated four median datasets (chlorophyll a, Chl a; total phosphorus, TP; and transparency) using multiple approaches to handling censored observations, including substituting fractions of the quantification limit (QL; dataset 1 = 1QL, dataset 2 = 0.5QL) and statistical methods for censored datasets (datasets 3-4) for approximately 100 Texas, USA reservoirs. Trend analyses of differences between dataset 1 and 3 medians indicated percent difference increased linearly above thresholds in percent censored data (%Cen). This relationship was extrapolated to estimate medians for site-parameter combinations with %Cen > 80%, which were combined with dataset 3 as dataset 4. Changepoint analysis of Chl a- and transparency-TP relationships indicated threshold differences up to 50% between datasets. Recursive analysis identified secondary thresholds in dataset 4. Threshold differences show that information introduced via substitution or missing due to limitations of statistical methods biased values, underestimated error, and inflated the strength of TP thresholds identified in datasets 1-3. Analysis of covariance identified differences in linear regression models relating transparency-TP between datasets 1, 2, and the more statistically robust datasets 3-4. Study findings identify high-risk scenarios for biased analytical outcomes when using substitution. These include high probability of median overestimation when %Cen > 50-60% for a single QL, or when %Cen is as low 16% for multiple QL's. Changepoint analysis was uniquely vulnerable to substitution effects when using medians from sites with %Cen > 50%. Linear regression analysis was less sensitive to substitution and missing data effects, but differences in model parameters for transparency cannot be discounted and could be magnified by log-transformation of the variables.

Comparing two periphyton collection methods commonly used for stream bioassessment and the development of numeric nutrient standards.

Periphyton is an important component of stream bioassessment, yet methods for quantifying periphyton biomass can differ substantially. A case study within the Arkansas Ozarks is presented to demonstrate the potential for linking chlorophyll-a (chl-a) and ash-free dry mass (AFDM) data sets amassed using two frequently used periphyton sampling protocols. Method A involved collecting periphyton from a known area on the top surface of variably sized rocks gathered from relatively swift-velocity riffles without discerning canopy cover. Method B involved collecting periphyton from the entire top surface of cobbles systematically gathered from riffle-run habitat where canopy cover was intentionally avoided. Chl-a and AFDM measurements were not different between methods (p = 0.123 and p = 0.550, respectively), and there was no interaction between method and time in the repeated measures structure of the study. However, significantly different seasonal distinctions were observed for chl-a and AFDM from all streams when data from the methods were combined (p < 0.001 and p = 0.012, respectively), with greater mean biomass in the cooler sampling months. Seasonal trends were likely the indirect results of varying temperatures. Although the size and range of this study were small, results suggest data sets collected using different methods may effectively be used together with some minor considerations due to potential confounding factors. This study provides motivation for the continued investigation of combining data sets derived from multiple methods of data collection, which could be useful in stream bioassessment and particularly important for the development of regional stream nutrient criteria for the southern Ozarks.

Seasonal Differences in Relationships between Nitrate Concentration and Denitrification Rates in Ditch Sediments Vegetated with Rice Cutgrass.

Increased application of nitrogen (N) fertilizers in agricultural systems contributes to significant environmental impacts, including eutrophication of fresh and coastal waters. Rice cutgrass [ (L.) Sw.] can significantly enhance denitrification potential in agricultural ditch sediments and potentially reduce N export from agricultural watersheds, but relationships with known drivers are not well understood. To address this, we examined effects of nitrate (NO) availability on dinitrogen gas (N) and NO fluxes seasonally. Net denitrification rates were measured as positive N fluxes from vegetated intact sediment cores using membrane inlet mass spectrometry (MIMS). We developed Michaelis-Menten models for N fluxes across NO gradients in the spring, summer, and fall seasons. Summer N models exhibited the highest (maximum amount of net N flux) and (concentration of NO in the overlying water at which the net N flux is half of ), with a maximum production of N of ∼20 mg N m h. Maximum percentage NO retention occurred at 1 mg NO L in the overlying water in all seasons, except summer where maximum retention persisted from 1 to 5 mg NO L. Denitrification rates were strongly correlated with NO uptake by vegetated sediments in spring ( = 0.94, < 0.0001) and summer ( = 0.97, < 0.0001), but low NO uptake in fall and winter resulted in virtually no net denitrification during these seasons. Our results indicate that vegetated ditch sediments may act as effective NO sinks during the growing season. Ditch sediments vegetated with cutgrass not only immobilized a significant fraction of NO entering them but also permanently removed as much as 30 to 40% of the immobilized NO through microbial denitrification.

It Takes Two to Tango: When and Where Dual Nutrient (N & P) Reductions Are Needed to Protect Lakes and Downstream Ecosystems.

Preventing harmful algal blooms (HABs) is needed to protect lakes and downstream ecosystems. Traditionally, reducing phosphorus (P) inputs was the prescribed solution for lakes, based on the assumption that P universally limits HAB formation. Reduction of P inputs has decreased HABs in many lakes, but was not successful in others. Thus, the "P-only" paradigm is overgeneralized. Whole-lake experiments indicate that HABs are often stimulated more by combined P and nitrogen (N) enrichment rather than N or P alone, indicating that the dynamics of both nutrients are important for HAB control. The changing paradigm from P-only to consideration of dual nutrient control is supported by studies indicating that (1) biological N fixation cannot always meet lake ecosystem N needs, and (2) that anthropogenic N and P loading has increased dramatically in recent decades. Sediment P accumulation supports long-term internal loading, while N may escape via denitrification, leading to perpetual N deficits. Hence, controlling both N and P inputs will help control HABs in some lakes and also reduce N export to downstream N-sensitive ecosystems. Managers should consider whether balanced control of N and P will most effectively reduce HABs along the freshwater-marine continuum.

Mitigating cyanobacterial harmful algal blooms in aquatic ecosystems impacted by climate change and anthropogenic nutrients.

Mitigating the global expansion of cyanobacterial harmful blooms (CyanoHABs) is a major challenge facing researchers and resource managers. A variety of traditional (e.g., nutrient load reduction) and experimental (e.g., artificial mixing and flushing, omnivorous fish removal) approaches have been used to reduce bloom occurrences. Managers now face the additional effects of climate change on watershed hydrologic and nutrient loading dynamics, lake and estuary temperature, mixing regime, internal nutrient dynamics, and other factors. Those changes favor CyanoHABs over other phytoplankton and could influence the efficacy of control measures. Virtually all mitigation strategies are influenced by climate changes, which may require setting new nutrient input reduction targets and establishing nutrient-bloom thresholds for impacted waters. Physical-forcing mitigation techniques, such as flushing and artificial mixing, will need adjustments to deal with the ramifications of climate change. Here, we examine the suite of current mitigation strategies and the potential options for adapting and optimizing them in a world facing increasing human population pressure and climate change.