Community Biological Ammonium Demand: A Conceptual Model for Cyanobacteria Blooms in Eutrophic Lakes.

Cyanobacterial harmful algal blooms (CyanoHABs) are enhanced by anthropogenic pressures, including excessive nutrient (nitrogen, N, and phosphorus, P) inputs and a warming climate. Severe eutrophication in aquatic systems is often manifested as non-N2-fixing CyanoHABs (e.g., Microcystis spp.), but the biogeochemical relationship between N inputs/dynamics and CyanoHABs needs definition. Community biological ammonium (NH4+) demand (CBAD) relates N dynamics to total microbial productivity and NH4+ deprivation in aquatic systems. A mechanistic conceptual model was constructed by combining nutrient cycling and CBAD observations from a spectrum of lakes to assess N cycling interactions with CyanoHABs. Model predictions were supported with CBAD data from a Microcystis bloom in Maumee Bay, Lake Erie, during summer 2015. Nitrogen compounds are transformed to reduced, more bioavailable forms (e.g., NH4+ and urea) favored by CyanoHABs. During blooms, algal biomass increases faster than internal NH4+ regeneration rates, causing high CBAD values. High turnover rates from cell death and remineralization of labile organic matter consume oxygen and enhance denitrification. These processes drive eutrophic systems to NH4+ limitation or colimitation under warm, shallow conditions and support the need for dual nutrient (N and P) control.

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.

Exploring long-term trends in microcystin toxin values associated with persistent harmful algal blooms in Grand Lake St Marys.

High external nutrient loads from agricultural runoff have led to persistent and highly toxic algal blooms in Grand Lake St Marys (GLSM) for decades. These pervasive blooms are concurrent with long-term (2009 - 2021) toxin and environmental monitoring, providing a robust weekly dataset for modeling microcystins. Median weekly microcystin concentrations (23.2 µg/L) routinely exceeded World Health Organization recreational limits (20 µg/L) for the study period (ranged 0.03 - 185.0 µg/L). Here, we used a Bayesian hierarchical dynamic linear model to hindcast weekly microcystin toxins using external nutrient loads from tributary data as well as internal lake nutrient and physicochemical concentrations. Overall, lake TN was the biggest driver of microcystin concentration in GLSM. Likewise, TN:TP was a strong negative driver of microcystin (i.e. low N:P ratios align with lower total microcystins), suggesting that N availability directly impacts toxins. External nutrient loading was positively related to microcystin during winter and spring; however, there was no relationship detected between toxin and external loading during summer or fall (particulate phosphorus exhibited the strongest signal but all external nutrients were unsurprisingly correlated). This lack of direct correlation on a weekly timescale between external loads and cyanobacterial toxins during the summer months likely results from nutrient saturation and reflects the importance of internal loading for bloom maintenance as supported by the correlation between in-lake TN and microcystin. Thus, management goals to reduce the highest biomass and toxins in the summer should focus on reduction of winter and spring external nutrient loads. Supporting this, both 2010 and 2021 had lower rain in the first half of the year (winter/spring), resulting in less loading, and experienced smaller/later low toxicity blooms. This suggests that, although internal nutrient loads are important for bloom maintenance, reduced external loads are an effective management strategy even in nutrient saturated systems such as GLSM.

Nitrate as a predictor of cyanobacteria biomass in eutrophic lakes in a climate change context.

We aimed to predict cyanobacteria biomass and nitrate (NO3-) concentrations in Lake Võrtsjärv, a large, shallow, and eutrophic lake in Estonia. We used a model chain based on the succession of a mechanistic (INCA-N) model and an empirical, generalized linear model. INCA-N model calibration and validation was performed with long term climate and catchment parameters. We constructed twelve scenarios as combinations of climate forcing from the Intergovernmental Panel on Climate Change (IPCC, 3 scenarios), land conversion (forest to agriculture, 2 scenarios), and fertilizer use (2 scenarios). Models predicted 46% of the variance of cyanobacteria biomass and 65% of that of NO3- concentrations. The model chain simulated that scenarios comprising both forest conversion to agricultural lands and a greater use of fertilizer per surface area unit would cause increases in lacustrine NO3- (up to twice the historical mean) and cyanobacteria biomass (up to a four-fold increase compared to the historical mean). The changes in NO3- concentrations and cyanobacteria biomass were more pronounced in low and moderate warming scenarios than in high warming scenarios because of increased denitrification rates in a warmer climate. Our findings show the importance of reducing anthropogenic pressures on lake catchments in order to reduce harmful pollutant and microalgae proliferation, and highlight the counterintuitive effects of multiple stressor interactions on lake functioning.

Hurricane Disturbance Stimulated Nitrification and Altered Ammonia Oxidizer Community Structure in Lake Okeechobee and St. Lucie Estuary (Florida).

Nitrification is an important biological link between oxidized and reduced forms of nitrogen (N). The efficiency of nitrification plays a key role in mitigating excess N in eutrophic systems, including those with cyanobacterial harmful algal blooms (cyanoHABs), since it can be closely coupled with denitrification and removal of excess N. Recent work suggests that competition for ammonium (NH4 +) between ammonia oxidizers and cyanoHABs can help determine microbial community structure. Nitrification rates and ammonia-oxidizing archaeal (AOA) and bacterial (AOB) community composition and gene abundances were quantified in Lake Okeechobee and St. Lucie Estuary in southern Florida (United States). We sampled during cyanobacterial (Microcystis) blooms in July 2016 and August 2017 (2 weeks before Hurricane Irma) and 10 days after Hurricane Irma made landfall. Nitrification rates were low during cyanobacterial blooms in Lake Okeechobee and St. Lucie Estuary, while low bloom conditions in St. Lucie Estuary coincided with greater nitrification rates. Nitrification rates in the lake were correlated (R 2 = 0.94; p = 0.006) with AOA amoA abundance. Following the hurricane, nitrification rates increased by an order of magnitude, suggesting that nitrifiers outcompeted cyanobacteria for NH4 + under turbid, poor light conditions. After Irma, AOA and AOB abundances increased in St. Lucie Estuary, while only AOB increased in Lake Okeechobee. AOA sequences clustered into three major lineages: Nitrosopumilales (NP), Nitrososphaerales (NS), and Nitrosotaleales (NT). Many of the lake OTUs placed within the uncultured and uncharacterized NS δ and NT ß clades, suggesting that these taxa are ecologically important along this eutrophic, lacustrine to estuarine continuum. After the hurricane, the AOA community shifted toward dominance by freshwater clades in St. Lucie Estuary and terrestrial genera in Lake Okeechobee, likely due to high rainfall and subsequent increased turbidity and freshwater loading from the lake into the estuary. AOB community structure was not affected by the disturbance. AOA communities were consistently more diverse than AOB, despite fewer sequences recovered, including new, unclassified, eutrophic ecotypes, suggesting a wider ecological biogeography than the oligotrophic niche originally posited. These results and other recent reports contradict the early hypothesis that AOB dominate ammonia oxidation in high-nutrient or terrestrial-influenced systems.

External Sources Inhibit Benthic Phosphorus Fluxes in the Lower Great Miami River, Southwest Ohio.

Human activities have increased nutrient loadings to aquatic ecosystems, especially during the past century. During low river flow in late summer and early fall, elevated concentrations of phosphorus (P) and nitrogen are present in the temperate Lower Great Miami River and contribute to its eutrophication. Although wastewater treatment plants are suspected of being major sources of P to the river, riverbed sediment has not been examined as an additional potential source of P. Benthic P fluxes were measured at 11 representative locations along the Lower Great Miami River during 3 sampling campaigns in late summer and early fall of 2015. Benthic fluxes of filtered total P (range, -1.6-12 mg m-2 d-1 ) were related inversely to filtered total P concentrations in river water (p = 0.002, r = -0.60). This relationship suggests that elevated P in river water inhibits mobilization from sediment, likely by minimizing the concentration gradient between porewater and overlying water. To effectively mitigate long-term effects of legacy P stored in Lower Great Miami River sediments, external sources must continue to be managed and discharges reduced to allow legacy P to be mobilized and flushed from the system. Reducing nutrient loading will help protect water quality in the Lower Great Miami River, in other comparable rivers, and in downstream aquatic habitats. Environ Toxicol Chem 2020;39:1517-1525. © 2020 SETAC.

Ammonium recycling supports toxic Planktothrix blooms in Sandusky Bay, Lake Erie: Evidence from stable isotope and metatranscriptome data.

Sandusky Bay, Lake Erie, receives high nutrient loadings (nitrogen and phosphorus) from the Sandusky River, which drains an agricultural watershed. Eutrophication and cyanobacterial harmful algal blooms (cyanoHABs) persist throughout summer. Planktothrix agardhii is the dominant bloom-forming species and the main producer of microcystins in Sandusky Bay. Non-N2 fixing cyanobacteria, such as Planktothrix and Microcystis, thrive on chemically reduced forms of nitrogen, such as ammonium (NH4+) and urea. Ammonium regeneration and potential uptake rates and total microbial community demand for NH4+ were quantified in Sandusky Bay. Potential NH4+ uptake rates in the light increased from June to August at all stations. Dark uptake rates also increased seasonally and, by the end of August, were on par with light uptake rates. Regeneration rates followed a similar pattern and were significantly higher in August than June. Ammonium uptake kinetics during a Planktothrix-dominated bloom in Sandusky Bay and a Microcystis-dominated bloom in Maumee Bay were also compared. The highest half saturation constant (Km) in Sandusky Bay was measured in June and decreased throughout the season. In contrast, Km values in Maumee Bay were lowest at the beginning of summer and increased in October. A significant increase in Vmax in Sandusky Bay was observed between July and the end of August, reflective of intense competition for depleted NH4+. Metatranscriptome results from Sandusky Bay show a shift from cyanophycin synthetase (luxury NH4+ uptake; cphA1) expression in early summer to cyanophycinase (intracellular N mobilization; cphB/cphA2) expression in August, supporting the interpretation that the microbial community is nitrogen-starved in late summer. Combined, our results show that, in late summer, when nitrogen concentrations are low, cyanoHABs in Sandusky Bay rely on regenerated NH4+ to support growth and toxin production. Increased dark NH4+ uptake late in summer suggests an important heterotrophic contribution to NH4+ depletion in the phycosphere. Kinetic experiments in the two bays suggest a competitive advantage for Planktothrix over Microcystis in Sandusky Bay due to its higher affinity for NH4+ at low concentrations.

Reduced forms of nitrogen are a driver of non-nitrogen-fixing harmful cyanobacterial blooms and toxicity in Lake Erie.

Western Lake Erie (WLE) experiences anthropogenic eutrophication and annual, toxic cyanobacterial blooms of non-nitrogen (N) fixing Microcystis. Numerous studies have shown that bloom biomass is correlated with an increased proportion of soluble reactive phosphorus loading from the Maumee River. Long term monitoring shows that the proportion of the annual Maumee River N load of non-nitrate N, or total Kjeldahl nitrogen (TKN), has also increased significantly (Spearman's ρ = 0.68, p = 0.001) over the last few decades and is also significantly correlated to cyanobacterial bloom biomass (Spearman's ρ = 0.64, p = 0.003). The ratio of chemically reduced N to oxidized N (TKN:NO3) concentrations was also compared to extracted chlorophyll and phycocyanin concentrations from all weekly sampling stations within WLE from 2009 to 2015. Both chlorophyll (Spearman's ρ = 0.657, p < 0.0001) and phycocyanin (Spearman's ρ = 0.714, p < 0.0001) were significantly correlated with TKN:NO3. This correlation between the increasing fraction of chemically reduced N from the Maumee River and increasing bloom biomass demonstrates the urgent need to control N loading, in addition to current P load reductions, to WLE and similar systems impacted by non-N-fixing, toxin-producing cyanobacteria.

Effects of silver nanoparticles on nitrification and associated nitrous oxide production in aquatic environments.

Silver nanoparticles (AgNPs) are the most common materials in nanotechnology-based consumer products globally. Because of the wide application of AgNPs, their potential environmental impact is currently a highly topical focus of concern. Nitrification is one of the processes in the nitrogen cycle most susceptible to AgNPs but the specific effects of AgNPs on nitrification in aquatic environments are not well understood. We report the influence of AgNPs on nitrification and associated nitrous oxide (N2O) production in estuarine sediments. AgNPs inhibited nitrification rates, which decreased exponentially with increasing AgNP concentrations. The response of nitrifier N2O production to AgNPs exhibited low-dose stimulation (<534, 1476, and 2473 µg liter-1 for 10-, 30-, and 100-nm AgNPs, respectively) and high-dose inhibition (hormesis effect). Compared with controls, N2O production could be enhanced by >100% at low doses of AgNPs. This result was confirmed by metatranscriptome studies showing up-regulation of nitric oxide reductase (norQ) gene expression in the low-dose treatment. Isotopomer analysis revealed that hydroxylamine oxidation was the main N2O production pathway, and its contribution to N2O emission was enhanced when exposed to low-dose AgNPs. This study highlights the molecular underpinnings of the effects of AgNPs on nitrification activity and demonstrates that the release of AgNPs into the environment should be controlled because they interfere with nitrifying communities and stimulate N2O emission.

Molecular evidence for sediment nitrogen fixation in a temperate New England estuary.

Primary production in coastal waters is generally nitrogen (N) limited with denitrification outpacing nitrogen fixation (N2-fixation). However, recent work suggests that we have potentially underestimated the importance of heterotrophic sediment N2-fixation in marine ecosystems. We used clone libraries to examine transcript diversity of nifH (a gene associated with N2-fixation) in sediments at three sites in a temperate New England estuary (Waquoit Bay, Massachusetts, USA) and compared our results to net sediment N2 fluxes previously measured at these sites. We observed nifH expression at all sites, including a site heavily impacted by anthropogenic N. At this N impacted site, we also observed mean net sediment N2-fixation, linking the geochemical rate measurement with nifH expression. This same site also had the lowest diversity (non-parametric Shannon = 2.75). At the two other sites, we also detected nifH transcripts, however, the mean N2 flux indicated net denitrification. These results suggest that N2-fixation and denitrification co-occur in these sediments. Of the unique sequences in this study, 67% were most closely related to uncultured bacteria from various marine environments, 17% to Cluster III, 15% to Cluster I, and only 1% to Cluster II. These data add to the growing body of literature that sediment heterotrophic N2-fixation, even under high inorganic nitrogen concentrations, may be an important yet overlooked source of N in coastal systems.

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.

Examining the impact of acetylene on N-fixation and the active sediment microbial community.

Here we examined the impact of a commonly employed method used to measure nitrogen fixation, the acetylene reduction assay (ARA), on a marine sediment community. Historically, the ARA technique has been broadly employed for its ease of use, in spite of numerous known artifacts. To gauge the severity of these effects in a natural environment, we employed high-throughput 16S rRNA gene sequencing to detect differences in acetylene-treated sediments vs. non-treated control sediments after a 7 h incubation. Within this short time period, significant differences were seen across all activity of microbes identified in the sediment, implying that the changes induced by acetylene occur quickly. The results have important implications for our understanding of marine nitrogen budgets. Moreover, because the ARA technique has been widely used in terrestrial and freshwater habitats, these results may be applicable to other ecosystems.

A shift in the archaeal nitrifier community in response to natural and anthropogenic disturbances in the northern Gulf of Mexico.

The Gulf of Mexico is affected by hurricanes and suffers seasonal hypoxia. The Deepwater Horizon oil spill impacted every trophic level in the coastal region. Despite their importance in bioremediation and biogeochemical cycles, it is difficult to predict the responses of microbial communities to physical and anthropogenic disturbances. Here, we quantify sediment ammonia-oxidizing archaeal (AOA) community diversity, resistance and resilience, and important geochemical factors after major hurricanes and the oil spill. Dominant AOA archetypes correlated with different geochemical factors, suggesting that different AOA are constrained by distinct parameters. Diversity was lowest after the hurricanes, showing weak resistance to physical disturbances. However, diversity was highest during the oil spill and coincided with a community shift, suggesting a new alternative stable state sustained for at least 1 year. The new AOA community was not significantly different from that at the spill site 1 year after the spill. This sustained shift in nitrifier community structure may be a result of oil exposure.