Cylindrospermopsis raciborskii-dominated harmful algae blooms have been reported globally in recent years. However, our understanding of the ecology of C. raciborskii in natural conditions is still poor. In this study, we collected the water samples from a C. raciborskii-blooming lake, Yilong Lake, in Yunnan province, China, and used both culture-dependent and culture-independent approaches to investigate their microbial communities and the interactions between C. raciborskii and the other bacteria. The composition and diversity of microbial communities were revealed with 16S rRNA gene high-throughput sequencing data analysis. Microbial co-occurrences analysis suggests C. raciborskii may have complex associations with other bacteria. Based on co-inoculation tests, we obtained 14 strains of bacterial strains from the water samples that exhibited either algicidal or promoting effects on a strain of C. raciborskii. Two bacterial isolates exhibited a consistent performance between co-occurrence analysis and experimental results. Effects of these bacteria-algae interspecies interactions on the bloom event are discussed. All these results may provide new insights into the C. raciborskii-dominated blooms and how its interspecies relationships with other bacteria may influence the bloom events in eutrophic waters throughout the world.

The cyanobacterial species Raphidiopsis raciborskii (Wołoszyńska) Aguilera et al. has a high invasiveness potential, which in less than a century leads to its cosmopolitan spread. In the temperate climate of Europe, R. raciborskii has been reported in many countries, but there is still a lack of detailed information about the current status of its distribution in lakes of Bulgaria, as a part of the southeastern range of its spread in Europe. We investigated the distribution of the species using data on the phytoplankton of 122 lakes surveyed during 13-years period (2009-2022). The species was found in 33 lakes (up to 324 m asl), and 14 new localities were registered during the studied period. The results reveal that the number of lakes with the presence of R. raciborskii (27 % of all research lakes) and its contribution to the total phytoplankton biomass, has increased significantly over the last decade. The species has successfully adapted and dominated the phytoplankton in 9 lakes, forming a bloom in 8 of them. The dominant position of R. raciborskii causes loss of species and functional diversity of phytoplankton and displaces the native bloom-forming cyanobacteria. Lakes with and without the species were compared based on the available data on bioclimatic and local environmental variables. Statistically significant differences were established with respect to water transparency, conductivity, maximum depth and maximum air temperature in the warmest month. Species distribution models (SDMs) were used to identify lakes in high risk of future invasion by R. raciborskii. The results of the SDMs implementation confirmed the high maximum air temperature and low water transparency to be important predictors of the occurrence of R. raciborskii in freshwater lakes in Bulgaria. In the areas with high summer temperatures the most suitable for R. raciborskii development were found to be shallow polymictic or medium deep lakes with small surface area and low water transparency. In areas with a suitable climate, the large, deep reservoirs with high transparency as well as macrophyte dominated lakes have a low probability of occurrence of R. raciborskii. Future colonization of lakes above 500 m asl (but most likely below 700 m asl) is also possible, especially in the conditions of global warming. SDMs account for climatic and biogeographic differences of lakes and could help in elucidating the underlying factors that control the occurrence and adaptation of R. raciborskii in a given area.

The proliferation of Raphidiopsis raciborskii blooms has sparked concerns regarding potential human exposure to heightened saxitoxins (STXs) levels. Thus, comprehending how environmental elements drive the proliferation of this STXs-producing species can aid in predicting human exposure risks. This study aimed to explore the link between cyanobacteria R. raciborskii, STXs cyanotoxins, and environmental factors in 37 public supply reservoirs in the tropical region and assess potential health hazards these toxins pose in the reservoir waters. A Structural Equation Model was used to assess the impact of environmental factors (water volume and physical and chemical variables) on R. raciborskii biomass and STXs levels. Furthermore, the potential risk of STXs exposure from consuming untreated reservoir water was evaluated. Lastly, the cumulative distribution function (CDF) of STXs across the reservoirs was computed. Our findings revealed a correlation between R. raciborskii biomass and STXs concentrations. Total phosphorus emerged as a critical environmental factor positively influencing species biomass and indirectly affecting STXs levels. pH significantly influenced STXs concentrations, indicating different factors influencing R. raciborskii biomass and STXs. Significantly, for the first time, the risk of STXs exposure was gauged using the risk quotient (HQ) for untreated water consumption from public supply reservoirs in Brazil\'s semi-arid region. Although the exposure risks were generally low to moderate, the CDF underscored the risk of chronic exposure due to low toxin concentrations in over 90% of samples. These outcomes emphasize the potential expansion of R. raciborskii in tropical settings due to increased phosphorus, amplifying waterborne STXs levels and associated intoxication risks. Thus, this study reinforces the importance of nutrient control, particularly phosphorus regulation, as a mitigation strategy against R. raciborskii blooms and reducing STXs intoxication hazards.

Two Raphidiopsis (=Cylindrospermopsis) raciborskii metagenome-assembled genomes (MAGs) were recovered from two freshwater metagenomic datasets sampled in 2011 and 2012 in Pampulha Lake, a hypereutrophic, artificial, shallow reservoir, located in the city of Belo Horizonte (MG), Brazil. Since the late 1970s, the lake has undergone increasing eutrophication pressure, due to wastewater input, leading to the occurrence of frequent cyanobacterial blooms. The major difference observed between PAMP2011 and PAMP2012 MAGs was the lack of the saxitoxin gene cluster in PAMP2012, which also presented a smaller genome, while PAMP2011 presented the complete sxt cluster and all essential proteins and clusters. The pangenome analysis was performed with all Raphidiopsis/Cylindrospermopsis genomes available at NCBI to date, with the addition of PAMP2011 and PAMP2012 MAGs (All33 subset), but also without the South American strains (noSA subset), and only among the South American strains (SA10 and SA8 subsets). We observed a substantial increase in the core genome size for the \'noSA\' subset, in comparison to \'All33\' subset, and since the core genome reflects the closeness among the pangenome members, the results strongly suggest that the conservation level of the essential gene repertoire seems to be affected by the geographic origin of the strains being analyzed, supporting the existence of a distinct SA clade. The Raphidiopsis pangenome comprised a total of 7943 orthologous protein clusters, and the two new MAGs increased the pangenome size by 11%. The pangenome based phylogenetic relationships among the 33 analyzed genomes showed that the SA genomes clustered together with 99% bootstrap support, reinforcing the metabolic particularity of the Raphidiopsis South American clade, related to its saxitoxin producing unique ability, while also indicating a different evolutionary history due to its geographic isolation.

As a tropical filamentous cyanobacterium, Raphidiopsis raciborskii has attracted much attention due to its expansion and toxin production. However, the mechanisms of its expansion to temperate regions have not been studied in detail. To address the potential strategies, the physiological and metabolomic profiles of R. raciborskii FACHB 1096 isolated from a temperate lake in China were determined and measured at different temperatures (10 °C, 15 °C, 20 °C, 25 °C, and 32 °C). The results demonstrated that temperature significantly changed cell viability, chlorophyll a content, specific growth rate, Chl a fluorescence, and filamentous shape of R. raciborskii. Low temperature decreased cell viability, specific growth rate, and photosynthetic efficiency, while the proportion of akinete and carbon fixation per unit cell were significantly increased compared with high temperature (32 °C). A constructed unimodal model indicated that filament length, cell volume, and cell length/width of R. raciborskii were significantly reduced in both high and low temperature environments. Under low-temperature conditions, R. raciborskii suffered different degrees of oxidative damage and produced corresponding antioxidant substances to resist oxidative stress, suggesting that low temperature changes the metabolic level of the cells, causing the cells to gradually switch from development to defense. Metabolomic data further confirmed that temperature change induced shifts in metabolic pathways in R. raciborskii, including starch and sucrose metabolic pathways, glutathione metabolic pathways, and the pentose phosphate pathways (PPP), as well as metabolic pathways related to the tricarboxylic acid (TCA) cycle. Our results indicated that the trade-offs of R. raciborskii cells among the growth, cell size, and metabolites can be significantly regulated by temperature, with broad implications for its global expansion in temperate waterbodies.

Water blooms caused by the invasive cyanobacterium Raphidiopsis raciborskii occur in many reservoirs in the tropical and subtropical regions of China. In recent decades, this species has spread rapidly to temperate regions. Phenotypic plasticity and climate warming are thought to promote the worldwide dispersion of R. raciborskii. However, investigations into the genetic and phenotypic diversities of this species have revealed significant intraspecific heterogeneity. In particular, competition between R. raciborskii and Microcystis aeruginosa was highly strain dependent. Although the concept of an ecotype was proposed to explain the heterogeneity of R. raciborskii strains with different geographic origins, microevolution is more reasonable for understanding the coexistence of different phenotypes and genotypes in the same environment. It has been suggested that intraspecific heterogeneity derived from microevolution is a strong driving force for the expansion of R. raciborskii. Additionally, temperature, nutrient fluctuations, and grazer disturbance are critical environmental factors that affect the population establishment of R. raciborskii in new environments. The present review provides new insights into the ecological mechanisms underlying the invasion of R. raciborskii in Chinese freshwater ecosystems.

Interactions between heterotrophic bacteria and cyanobacteria regulate the structure and function of aquatic ecosystems and are thus crucial for the prediction and management of cyanobacterial blooms in relation to water security. Currently, abundant bacterial species are of primary concern, while the role of more diverse and copious rare species remains largely unknown. Using a dilution-to-extinction approach, rare bacterial species from reservoir water were co-cultured with the bloom-forming cyanobacterium Raphidiopsis raciborskii in the lab to explore their interactions by using Phyto-PAM and 16S rRNA gene high-throughput sequencing. We found that a ≤1000-fold bacterial dilution led to bacteria control of the growth and photosynthesis of R. raciborskii. Moreover, the bacterial community compositions in the low-dilution groups were clearly diverged from the high-dilution groups. Importantly, rare species changed dramatically in the low-dilution groups, resulting in lower phylogenetic diversity and narrower niche width. The network complexity and compositional stability of bacterial communities decreased in the low-dilution groups. Collectively, our results suggest that rare bacterial species inhibit R. raciborskii growth and photosynthesis through microbial interactions mediated by species coexistence and interaction mechanisms. Our study provides new knowledge of the ecological role of rare bacteria and offers new perspectives for understanding the outbreak and regression of R. raciborskii blooms.

The succession of dominant species always occurs during cyanobacterial blooms because there are certain conditions for cyanobacterial blooms formed by different cyanobacteria; this results in more uncertain and complex effects in cyanobacterial blooms. However, the succession pattern and consequences of dominant species and its driving factors have not received enough attention during cyanobacteria blooms. In this study, the phytoplankton community characteristics and water environment factors of Nanpeng Reservoir, a drinking water source in Chongqing, were monitored and analyzed from April to September 2018. The results showed that:① a total of 108 species of phytoplankton belonging to 59 genera and 8 phyla were identified in Nanpeng Reservoir. Of this, 13 species of 4 phyla were identified as dominant species, among which the dominance index of Cylindrospermopsis raciborskii was the highest, followed by that of Pseudoanabaena sp. ② The most dominant cyanobacteria were Pseudoanabaena and Cylindrospermopsis in May and July, respectively, in which cyanobacteria density peaked, whereas the Shannon-Weiner diversity and Pielou evenness were significantly lower than those in the other months. ③ NMDS results showed that the correlation between Cylindrospermopsis or Pseudoanabaena and the ambient phytoplankton community was 0.58 and 0.48, respectively. Moreover, the VPA results showed that 47.51% of the community variation could be explained by environmental factors, and only 12.04% and 12.74% of variation in community composition could be explained by Cylindrospermopsis and Pseudoanabaena, respectively. ④ The abundance of Cylindrospermopsis was significantly positively affected by WT, pH, and RUEN and negatively affected by SD and RUEP. However, the abundance of Pseudoanabaena was significantly positively affected by permanganate index and negatively affected by EC and DO. These results suggested that both dominating cyanobacteria had significant effects on the surrounding phytoplankton community. Relative to that of Pseudoanabaena, however, Cylindrospermopsis had a more obvious impact on the aquatic ecosystem. Moreover, the synergistic effect of N limitation and warming of the water column may have caused the replacement of Pseudoanabaena with Cylindrospermopsis to form a dominant population.

In recent years, Cylindrospermopsis raciborskii blooms have been widely found worldwide. Topics dealing with the mitigation of C. raciborskii bloom is of great importance for toxins produced could threaten public health. The paper first investigated C. raciborskii dynamics over three years following sediment dredging in a shallow eutrophic Lake Dongqian (China). Based on rpoC1 gene copies, C. raciborskii bloom formed with average density of 1.30 × 106 cells/L on July 2009. One year later after sediment dredging, C. raciborskii cell density decreased below 1.17 × 105 cells/L or under detected limits during summer days on 2010. While two years later, the C. raciborskii bloom period was returned with markedly increased cell density reaching up to 4.15 × 107 cells/L on October 2011, and the maximum peak density was shown at 20.3 °C that was much lower than reported optimal growth temperature. Inferred from Spearman correlation analysis, linear regression showed C. raciborskii density was significant and positive with pH and SD, whereas they were significant and negative with TP and DO. Multiple regression analysis further demonstrated that TN, TP, SRP, pH and DO provided the best model and explained 53.1% of the variance in C. raciborskii dynamics. The approaches managing nutrients reduction might not control C. raciborskii bloom as extremely low TN (avg. 0.18 mg/L) and TP concentrations (avg. 0.05 mg/L) resulted in the highest C. raciborskii cell density after sediment dredging.

The concentration of coloured terrestrial dissolved organic matter (tDOM) from vegetation appears to be increasing in lakes in some regions of the world, leading to the term brownification. The light attenuating effect of coloured tDOM on phytoplankton growth has been a major focus of attention, but the phytotoxic effects of tDOM, particularly on cyanobacterial blooms, are less well understood. This mesocosm study tested whether coloured tDOM, leached from the leaves of a Eucalyptus tree species, inhibited a naturally occurring bloom of the toxic cyanobacterium, Raphidiopsis raciborskii, in a reservoir over a 10 day period. The study found that tDOM leachate, measured as dissolved organic carbon (DOC), inhibited photosynthesis and growth of both R. raciborskii, as well as species present at lower densities, i.e. other cyanobacteria and diatoms. However, the effect was greater at higher tDOM input loads. The photosynthetic yield (Fv/Fm) of cyanobacteria decreased rapidly in treatments with 5.9 and 25 mg L-1 DOC addition, compared to the control (reservoir water with background DOC concentration of 6.85 ± 1.09 mg L-1). tDOM had no measurable effect in the 2 and 3.3 mg L-1 DOC addition treatments. By day 5, cell densities of cyanobacteria, including R. raciborskii, and diatoms, in treatments with 5.9 and 25 mg L-1 DOC addition were significantly lower than the control with no tDOM addition, and this effect continued throughout the experiment. This is despite the leachate addition increasing phosphate concentrations which counteracted the low background concentrations of phosphate. Light attenuation and dissolved oxygen (DO) levels were also affected by the tDOM addition, but were only significantly lower in the 25 mg L-1 DOC treatment compared with the control. DOC, dissolved organic nitrogen (DON) and dissolved organic phosphorus (DOP) concentrations all decreased in the tDOM addition treatments over the first 3 days, as the microbial cell densities increased. The components of the tDOM that decreased over time were determined by 1H NMR spectroscopy in the 25 mg L-1 DOC treatment. After 5 d, the relative concentrations of fatty acids, sugars and gallic acid decreased by around 60%, while concentrations of flavonoids and myo-inositol decreased by 45 and 35% respectively. This study suggests that phytotoxic compounds in tDOM can suppress cyanobacterial blooms, despite the increased nutrient inputs. This has implications for predicting the future likelihood of cyanobacterial blooms in lakes and reservoirs with climate-change driven changes in flow events, and other changes in the amount and types of vegetation cover. Revegetation of riparian zones, resulting in increased tDOM into waterways, may also be beneficial in reducing cyanobacterial blooms.