Coupling of cylindrospermopsin and pho-harboring Verrucomicrobia supports the formation of Raphidiopsis blooms in low-phosphorus waters.

Cylindrospermopsin (CYN) can induce phytoplankton community to secrete alkaline phosphatase (ALP), which is one of the important strategies for the bloom-forming cyanobacterium Raphidiopsis to thrive in extremely low-phosphorus (P) waters. However, how bacterioplankton community, another major contributor to ALPs in waters, couples to Raphidiopsis through CYN, and the role of this coupling in supporting the dominance of Raphidiopsis in nature remain largely unknown. Here, we conducted microcosm experiments to address this knowledge gap, using a combination of differential filtration-based and metagenomics-based methods to identify the sources of ALPs. We found that, compared with algal-derived ALPs, bacteria-derived ALPs exhibited a more pronounced and sensitive response to CYN. This response to CYN was enhanced under low-P conditions. Interestingly, we found that Verrucomicrobia made the largest contribution to the total abundance of pho genes, which encode ALPs. Having high gene abundance of the CYN-sensing PI3K-AKT signaling pathway, Verrucomicrobia's proportion increased with higher concentrations of CYN under low-P conditions, thereby explaining the observed increase in pho gene abundance. Compared with other cyanobacterial genera, Raphidiopsis had a higher abundance of the pst gene. This suggests that Raphidiopsis exhibited a greater capacity to uptake the inorganic P generated by ALPs secreted by other organisms. Overall, our results reveal the mechanism of CYN-induced ALP secretion and its impact on planktonic P-cycling, and provide valuable insights into the role of CYN in supporting the formation of Raphidiopsis blooms.

Mitigating the global expansion of harmful cyanobacterial blooms: Moving targets in a human- and climatically-altered world.

Cyanobacterial harmful algal blooms (CyanoHABs) are a major threat to human and environmental health. As global proliferation of CyanoHABs continues to increase in prevalence, intensity, and toxicity, it is important to identify and integrate the underlying causes and controls of blooms in order to develop effective short- and long-term mitigation strategies. Clearly, nutrient input reductions should receive high priority. Legacy effects of multi-decadal anthropogenic eutrophication have altered limnetic systems such that there has been a shift from exclusive phosphorus (P) limitation to nitrogen (N) limitation and N and P co-limitation. Additionally, climate change is driving CyanoHAB proliferation through increasing global temperatures and altered precipitation patterns, including more extreme rainfall events and protracted droughts. These scenarios have led to the "perfect storm scenario"; increases in pulsed nutrient loading events, followed by persistent low-flow, long water residence times, favoring bloom formation and proliferation. To meet the CyanoHAB mitigation challenge, we must: (1) Formulate watershed and airshed-specific N and P input reductions on a sliding scale to meet anthropogenic and climatic forcings. (2) Develop CyanoHAB management strategies that incorporate current and anticipated climatic changes and extremes. (3) Make nutrient management strategies compatible with other physical-chemical-biological mitigation approaches, such as altering freshwater flow and flushing, dredging, chemical applications, introduction of selective grazers, etc. (4) Target CyanoHAB toxin production and developing management approaches to reduce toxin production. (5) Develop broadly applicable long-term strategies that incorporate the above recommendations.

Improvement in municipal wastewater treatment alters lake nitrogen to phosphorus ratios in populated regions.

Large-scale and rapid improvement in wastewater treatment is common practice in developing countries, yet this influence on nutrient regimes in receiving waterbodies is rarely examined at broad spatial and temporal scales. Here, we present a study linking decadal nutrient monitoring data in lakes with the corresponding estimates of five major anthropogenic nutrient discharges in their surrounding watersheds over time. Within a continuous monitoring dataset covering the period 2008 to 2017, we find that due to different rates of change in TN and TP concentrations, 24 of 46 lakes, mostly located in China's populated regions, showed increasing TN/TP mass ratios; only 3 lakes showed a decrease. Quantitative relationships between in-lake nutrient concentrations (and their ratios) and anthropogenic nutrient discharges in the surrounding watersheds indicate that increase of lake TN/TP ratios is associated with the rapid improvement in municipal wastewater treatment. Due to the higher removal efficiency of TP compared with TN, TN/TP mass ratios in total municipal wastewater discharge have continued to increase from a median of 10.7 (95% confidence interval, 7.6 to 15.1) in 2008 to 17.7 (95% confidence interval, 13.2 to 27.2) in 2017. Improving municipal wastewater collection and treatment worldwide is an important target within the 17 sustainable development goals set by the United Nations. Given potential ecological impacts on biodiversity and ecosystem function of altered nutrient ratios in wastewater discharge, our results suggest that long-term strategies for domestic wastewater management should not merely focus on total reductions of nutrient discharges but also consider their stoichiometric balance.

Effects of phosphorus availability and phosphorus utilization behavior of Microcystis aeruginosa on its adaptation capability to ultraviolet radiation.

Phosphorus (P) plays a critical role in eutrophication and algal growth; therefore, improving our understanding of the impact of P is essential to control harmful algal blooms. In this study, Microcystis aeruginosa was treated with 5-h ambient irradiation in the medium with different dissolved inorganic P (DIP) concentrations, DIP-free, moderate-DIP, and high-DIP, to explore its growth and other physiological responses. Compared to photosynthetically active radiation (PAR), UV-A (320-400 nm) and UV-B (280-320 nm) radiation had inhibitive effects on the photosynthesis and growth of M. aeruginosa, while high P availability could alleviate or eliminate the negative effects of UV radiation. The photosynthetic parameters had a minimum reduction and quickly recovered after re-inoculation under high-DIP conditions. Confirmed by SEM, photosynthetic pigments, the generation of reactive oxygen species (ROS), superoxide dismutase (SOD) activity and other methods, ambient UV radiation exerted oxidative stresses rather than direct lethal effects on M. aeruginosa. Photosynthetic parameters indicated that algal UV-adaptation processes could include decreasing photo-induced damages and increasing self-repair efficiency. The P acquired by M. aeruginosa cells can have two function, which included alleviating UV-induced negative effects and sustaining algal growth. Consequently, UV-adaptation processes of M. aeruginosa resulted in an elevated demand for DIP, which resulted to increased P uptake rates and cellular P quota under moderate and high-DIP conditions. Therefore, the production of carotenoid and phycocyanin, and SOD activity increased under UV stress, leading to a better adaptation capability of M. aeruginosa and decreased negative effects of UV radiation on its growth. Overall, our findings demonstrated the significant interactive effects of P enrichment and irradiation on typical cyanobacteria, and the strong adaptation capability of M. aeruginosa in the eutrophic UV-radiated waters.

Feedback Regulation between Aquatic Microorganisms and the Bloom-Forming Cyanobacterium Microcystis aeruginosa.

The frequency and intensity of cyanobacterial blooms are increasing worldwide. Interactions between toxic cyanobacteria and aquatic microorganisms need to be critically evaluated to understand microbial drivers and modulators of the blooms. In this study, we applied 16S/18S rRNA gene sequencing and metabolomics analyses to measure the microbial community composition and metabolic responses of the cyanobacterium Microcystis aeruginosa in a coculture system receiving dissolved inorganic nitrogen and phosphorus (DIP) close to representative concentrations in Lake Taihu, China. M. aeruginosa secreted alkaline phosphatase using a DIP source produced by moribund and decaying microorganisms when the P source was insufficient. During this process, M. aeruginosa accumulated several intermediates in energy metabolism pathways to provide energy for sustained high growth rates and increased intracellular sugars to enhance its competitive capacity and ability to defend itself against microbial attack. It also produced a variety of toxic substances, including microcystins, to inhibit metabolite formation via energy metabolism pathways of aquatic microorganisms, leading to a negative effect on bacterial and eukaryotic microbial richness and diversity. Overall, compared with the monoculture system, the growth of M. aeruginosa was accelerated in coculture, while the growth of some cooccurring microorganisms was inhibited, with the diversity and richness of eukaryotic microorganisms being more negatively impacted than those of prokaryotic microorganisms. These findings provide valuable information for clarifying how M. aeruginosa can potentially modulate its associations with other microorganisms, with ramifications for its dominance in aquatic ecosystems.IMPORTANCE We measured the microbial community composition and metabolic responses of Microcystis aeruginosa in a microcosm coculture system receiving dissolved inorganic nitrogen and phosphorus (DIP) close to the average concentrations in Lake Taihu. In the coculture system, DIP is depleted and the growth and production of aquatic microorganisms can be stressed by a lack of DIP availability. M. aeruginosa could accelerate its growth via interactions with specific cooccurring microorganisms and the accumulation of several intermediates in energy metabolism-related pathways. Furthermore, M. aeruginosa can decrease the carbohydrate metabolism of cooccurring aquatic microorganisms and thus disrupt microbial activities in the coculture. This also had a negative effect on bacterial and eukaryotic microbial richness and diversity. Microcystin was capable of decreasing the biomass of total phytoplankton in aquatic microcosms. Overall, compared to the monoculture, the growth of total aquatic microorganisms is inhibited, with the diversity and richness of eukaryotic microorganisms being more negatively impacted than those of prokaryotic microorganisms. The only exception is M. aeruginosa in the coculture system, whose growth was accelerated.

Mitigating the Expansion of Harmful Algal Blooms Across the Freshwater-to-Marine Continuum.

Anthropogenic nutrient overenrichment, coupled with rising temperatures, and an increasing frequency of extreme hydrologic events (storms and droughts) are accelerating eutrophication and promoting the expansion of harmful algal blooms (HABs) across the freshwater-to-marine continuum. All HABs-with a focus here on cyanobacterial blooms-pose serious consequences for water supplies, fisheries, recreational uses, tourism, and property values. As nutrient loads grow in watersheds, they begin to compound the effects of legacy stores. This has led to a paradigm shift in our understanding of how nutrients control eutrophication and blooms. Phosphorus (P) reductions have been traditionally prescribed exclusively for freshwater systems, while nitrogen (N) reductions were mainly stressed for brackish and coastal waters. However, because most systems are hydrologically interconnected, single nutrient (e.g., P only) reductions upstream may not necessarily reduce HAB impacts downstream. Reducing both N and P inputs is the only viable nutrient management solution for long-term control of HABs along the continuum. This article highlights where paired physical, chemical, or biological controls may improve beneficial uses in the short term, and offers management strategies that should be enacted across watershed scales to combat the global expansion of HABs across geographically broad freshwater-to-marine continua.

Controlling cyanobacterial harmful blooms in freshwater ecosystems.

Cyanobacteria's long evolutionary history has enabled them to adapt to geochemical and climatic changes, and more recent human and climatic modifications of aquatic ecosystems, including nutrient over-enrichment, hydrologic modifications, and global warming. Harmful (toxic, hypoxia-generating, food web altering) cyanobacterial bloom (CyanoHAB) genera are controlled by the synergistic effects of nutrient (nitrogen and phosphorus) supplies, light, temperature, water residence/flushing times, and biotic interactions. Accordingly, mitigation strategies are focused on manipulating these dynamic factors. Strategies based on physical, chemical (algaecide) and biological manipulations can be effective in reducing CyanoHABs. However, these strategies should invariably be accompanied by nutrient (both nitrogen and phosphorus in most cases) input reductions to ensure long-term success and sustainability. While the applicability and feasibility of various controls and management approaches is focused on freshwater ecosystems, they will also be applicable to estuarine and coastal ecosystems. In order to ensure long-term control of CyanoHABs, these strategies should be adaptive to climatic variability and change, because nutrient-CyanoHAB thresholds will likely be altered in a climatically more-extreme world.

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.

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.

Duelling 'CyanoHABs': unravelling the environmental drivers controlling dominance and succession among diazotrophic and non-N2 -fixing harmful cyanobacteria.

Eutrophication often manifests itself by increased frequencies and magnitudes of cyanobacterial harmful algal blooms (CyanoHABs) in freshwater systems. It is generally assumed that nitrogen-fixing cyanobacteria will dominate when nitrogen (N) is limiting and non-N2 fixers dominate when N is present in excess. However, this is rarely observed in temperate lakes, where N2 fixers often bloom when N is replete, and non-fixers (e.g. Microcystis) dominate when N concentrations are lowest. This review integrates observations from previous studies with insights into the environmental factors that select for CyanoHAB groups. This information may be used to predict how nutrient reduction strategies targeting N, phosphorus (P) or both N and P may alter cyanobacterial community composition. One underexplored concern is that as N inputs are reduced, CyanoHABs may switch from non-N2 fixing to diazotrophic taxa, with no net improvement in water quality. However, monitoring and experimental observations indicate that in eutrophic systems, minimizing both N and P loading will lead to the most significant reductions in total phytoplankton biomass without this shift occurring, because successional patterns appear to be strongly driven by physical factors, including temperature, irradiance and hydrology. Notably, water temperature is a primary driver of cyanobacterial community succession, with warming favouring non-diazotrophic taxa.

Green algal over cyanobacterial dominance promoted with nitrogen and phosphorus additions in a mesocosm study at Lake Taihu, China.

Enrichment of waterways with nitrogen (N) and phosphorus (P) has accelerated eutrophication and promoted cyanobacterial blooms worldwide. An understanding of whether cyanobacteria maintain their dominance under accelerated eutrophication will help predict trends and provide rational control measures. A mesocosm experiment was conducted under natural light and temperature conditions in Lake Taihu, China. It revealed that only N added to lake water promoted growth of colonial and filamentous cyanobacteria (Microcystis, Pseudoanabaena and Planktothrix) and single-cell green algae (Cosmarium, Chlorella, and Scenedesmus). Adding P alone promoted neither cyanobacteria nor green algae significantly. N plus P additions promoted cyanobacteria and green algae growth greatly. The higher growth rates of green algae vs. cyanobacteria in N plus P additions resulted in the biomass of green algae exceeding that of cyanobacteria. This indicates that further enrichment with N plus P in eutrophic water will enhance green algae over cyanobacterial dominance. However, it does not mean that eutrophication problems will cease. On the contrary, the risk will increase due to increasing total phytoplankton biomass.

Facultative diazotrophy increases Cylindrospermopsis raciborskii competitiveness under fluctuating nitrogen availability.

Relative fitness of three bloom-forming and potentially toxic cyanobacteria from the subtropical St. John's River, Florida was investigated under a range of nutrient conditions, during a bloom dominated by Cylindrospermopsis raciborskii. Nitrogen (N) was the primary nutrient limiting phytoplankton primary productivity and biomass. Phytoplankton biomass was also enhanced by phosphorus (P) added either alone or jointly with N, suggesting different components of the phytoplankton experienced distinct nutrient limitations. Based on quantitative PCR, the diazotrophic cyanobacteria Anabaena sp. and C. raciborskii were responsible for the primary production response to P additions, while the nondiazotrophic Microcystis aeruginosa appeared to benefit from N released from the diazotrophs. Cylindrospermopsis raciborskii maintained high net growth rates under diazotrophic and nondiazotrophic conditions, while Anabaena sp. growth was significantly reduced under DIN enrichment. C. raciborskii appears to be a generalist with regard to N source, a lifestyle traditionally not considered a viable ecological strategy among diazotrophs. Using facultative diazotrophy, C. raciborskii gains a growth advantage under fluctuating DIN conditions, such as systems that are under the influence of anthropogenic N loading events. The described niche differentiation may be a key factor explaining the recent global expansion of C. raciborskii.

Rationale for control of anthropogenic nitrogen and phosphorus to reduce eutrophication of inland waters.

Concentrations of phosphorus and nitrogen in surface waters are being regulated in the United States and European Union. Human activity has raised the concentrations of these nutrients, leading to eutrophication of inland waters, which causes nuisance growth of algae and other aquatic plants. Control of phosphorus often has had the highest priority because of its presumed leading role in limiting development of aquatic plant biomass. Experimental evidence shows, however, that nitrogen is equally likely to limit growth of algae and aquatic plants in inland waters, and that additions of both nutrients cause substantially more algal growth than either added alone. A dual control strategy for N and P will reduce transport of anthropogenic nitrogen through drainage networks to aquatic ecosystems that may be nitrogen limited. Control of total phosphorus in effluents is feasible and is increasingly being required by regulations. The control strategy for nitrogen in effluents is more difficult, but could be made more feasible by recognition that a substantial portion of dissolved organic nitrogen is not bioavailable; regulation should focus on bioavailable N (nitrate, ammonium, and some dissolved organic nitrogen) rather than total N. Regulation of both N and P also is essential for nonpoint sources.

Controlling harmful cyanobacterial blooms in a world experiencing anthropogenic and climatic-induced change.

Harmful (toxic, food web altering, hypoxia generating) cyanobacterial algal blooms (CyanoHABs) are proliferating world-wide due to anthropogenic nutrient enrichment, and they represent a serious threat to the use and sustainability of our freshwater resources. Traditionally, phosphorus (P) input reductions have been prescribed to control CyanoHABs, because P limitation is widespread and some CyanoHABs can fix atmospheric nitrogen (N(2)) to satisfy their nitrogen (N) requirements. However, eutrophying systems are increasingly plagued with non N(2) fixing CyanoHABs that are N and P co-limited or even N limited. In many of these systems N loads are increasing faster than P loads. Therefore N and P input constraints are likely needed for long-term CyanoHAB control in such systems. Climatic changes, specifically warming, increased vertical stratification, salinization, and intensification of storms and droughts play additional, interactive roles in modulating CyanoHAB frequency, intensity, geographic distribution and duration. In addition to having to consider reductions in N and P inputs, water quality managers are in dire need of effective tools to break the synergy between nutrient loading and hydrologic regimes made more favorable for CyanoHABs by climate change. The more promising of these tools make affected waters less hospitable for CyanoHABs by 1) altering the hydrology to enhance vertical mixing and/or flushing and 2) decreasing nutrient fluxes from organic rich sediments by physically removing the sediments or capping sediments with clay. Effective future CyanoHAB management approaches must incorporate both N and P loading dynamics within the context of altered thermal and hydrologic regimes associated with climate change.

Controlling harmful cyanobacterial blooms in a hyper-eutrophic lake (Lake Taihu, China): the need for a dual nutrient (N & P) management strategy.

Harmful cyanobacterial blooms, reflecting advanced eutrophication, are spreading globally and threaten the sustainability of freshwater ecosystems. Increasingly, non-nitrogen (N(2))-fixing cyanobacteria (e.g., Microcystis) dominate such blooms, indicating that both excessive nitrogen (N) and phosphorus (P) loads may be responsible for their proliferation. Traditionally, watershed nutrient management efforts to control these blooms have focused on reducing P inputs. However, N loading has increased dramatically in many watersheds, promoting blooms of non-N(2) fixers, and altering lake nutrient budgets and cycling characteristics. We examined this proliferating water quality problem in Lake Taihu, China's 3rd largest freshwater lake. This shallow, hyper-eutrophic lake has changed from bloom-free to bloom-plagued conditions over the past 3 decades. Toxic Microcystis spp. blooms threaten the use of the lake for drinking water, fisheries and recreational purposes. Nutrient addition bioassays indicated that the lake shifts from P limitation in winter-spring to N limitation in cyanobacteria-dominated summer and fall months. Combined N and P additions led to maximum stimulation of growth. Despite summer N limitation and P availability, non-N(2) fixing blooms prevailed. Nitrogen cycling studies, combined with N input estimates, indicate that Microcystis thrives on both newly supplied and previously-loaded N sources to maintain its dominance. Denitrification did not relieve the lake of excessive N inputs. Results point to the need to reduce both N and P inputs for long-term eutrophication and cyanobacterial bloom control in this hyper-eutrophic system.

Morphological and genetic evidence that the cyanobacterium Lyngbya wollei (Farlow ex Gomont) Speziale and Dyck encompasses at least two species.

Dense blooms of the cyanobacterium Lyngbya wollei are increasingly responsible for declining water quality and habitat degradation in numerous springs, rivers, and reservoirs. This research represents the first molecular phylogenetic analysis of L. wollei in comparison with the traditional morphological characterization of this species. Specimens were collected from several springs in Florida and a reservoir in North Carolina. Segments of the small-subunit (SSU) rRNA and nifH genes were PCR amplified, cloned, and sequenced. The phylogenetic analysis of the SSU rRNA gene revealed sequences that fell into three distinct subclusters, each with >97% sequence similarity. These were designated operational taxonomic unit 1 (OTU1), OTU2, and OTU3. Similarly, the nifH sequences fell into three distinct subclusters named S1, S2, and S3. When either bulk samples or individual filaments were analyzed, we recovered OTU1 with S1, OTU2 with S2, and OTU3 with S3. The coherence between the three SSU rRNA gene and nifH subclusters was consistent with genetically distinct strains or species. Cells associated with subclusters OTU3 and S3 were significantly wider and longer than those associated with other subclusters. The combined molecular and morphological data indicate that the species commonly identified as L. wollei in the literature represents two or possibly more species. Springs containing OTU3 and S3 demonstrated lower ion concentrations than other collection sites. Geographical locations of Lyngbya subclusters did not correlate with residual dissolved inorganic nitrogen or phosphorus concentrations. This study emphasizes the need to complement traditional identification with molecular characterization to more definitively detect and characterize harmful cyanobacterial species or strains.