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This study investigates the capability of a Bacillus flexus strain isolated from decayed cyanobacterial blooms for the bioremediation of Cylindrospermopsis raciborskii and cylindrospermopsin (CYN) toxin. The algicidal activity of this strain was tested by co-cultivation with C. raciborskii cultures. CYN biodegradation was investigated in the presence of living and heat-inactivated bacterial cells or bacterial filtrate. Living bacterial cells inhibited C. raciborskii growth after 2 days of incubation with complete cell death at day 5. Bacterial filtrate caused a rapid reduction in C. raciborskii growth at the first day, with complete cell lysis at day 3. Only living cells of SSZ01 caused reduction in CYN released into the medium during the bacterial decay of C. raciborskii cells. The biodegradation rate of CYN by SSZ01 relied on initial toxin concentrations. The highest rate (42 μg CYN L−1 day−1) was obtained at the higher initial concentration (300 μg L−1), and the lowest (4μg CYN L−1 day−1) was at lower concentration (50 μg L−1). These results suggest that this bacterial strain could be employed to bioremediate cyanobacterial blooms in freshwaters. Also, the application of this bacterium in slow sand filters would give possibilities for degradation and bioremediation of cyanotoxins in drinking water treatment plants.

Impact of macrophyte Lemna trisulca on the growth rate and synthesis of cylindrospermopsin (CYN) by cyanobacterium Raphidiopsis raciborskii was determined. The presence of L. trisulca inhibited the biomass accumulation of the cyanobacterium by 25% compared to the control during co-cultivation. The simultaneous cultivation of these organisms slightly affected the inhibition of macrophyte growth rate by 5.5% compared to the control. However, no morphological changes of L. trisulca after incubation with cyanobacteria were observed. It was also shown that the long-term (35 days) co-cultivation of R. raciborskii and L. trisulca led to a decrease in CYN concentration in media and cyanobacterial cells by 32 and 38%, respectively, compared to the values obtained for independent cultivation of cyanobacterium. Excessive absorption of phosphate ions by L. trisulca from the medium compared to nitrate ions led to a significant increase in the nitrate:phosphate ratio in the media, which inhibits the development of cyanobacterium. The obtained results indicate that L. trisulca in the natural environment may affect the physiology of cyanobacteria. The presented study is the first assessment of the allelopathic interaction of macrophyte and R. raciborskii.

Phosphorus (P) is responsible for algal growth and the structural changes in algal communities. Therefore, it is essential to know whether the different phosphorus availability to different algae can change the community structure. In this study, the interspecific competition was investigated at two bloom-forming cyanobacterium, Cylindrospermopsis raciborskii and Microcystis aeruginosa, when both were treated with five different phosphate compounds, including K2HPO4, β-glycerol phosphate, (2-aminoethyl)-phosphinic acid, glyphosate, and P-free. The results of mono-culture experiments showed that the two species could utilize the dissolved organic phosphorus (DOP) and K2HPO4 (DIP) as the sole P resource. Moreover, the specific growth rates and the endogenous alkaline phosphatase activity in M. aeruginosa cells were much lower than those in C. raciborskii under DOP and DIP treatments. In the co-cultured experiments, however, a significant biomass increase in C. raciborskii was observed in all experimental P treatments, except for glyphosate, regardless of its initial cell density proportion. A 31.8–63.4% increase in cell number of C. raciborskii was found after incubated into K2HPO4, while the highest biomass of mixed samples, 17.72 × 106 cell mL−1, was observed in the (2-aminoethyl)-phosphinic acid treatment (50C50M). Additionally, higher specific growth rate was also found in C. raciborskii when compared with M. aeruginosa under P-free; the increasing proportion of C. raciborskii were 29.1% (50C50M), 16.4% (75C25M), and 36.7% (25C75M), respectively. When the mixed samples were co-cultivated under glyphosate, C. raciborskii cells appeared to be depressed, whereas the cell density of M. aeruginosa increased rapidly. The findings indicated that an excellent P competition might give some advantages for C. raciborskii dominance in natural waters with DIP limitation or DOP abundance.

Koka Reservoir in Ethiopia has been severely impacted by water quality deterioration associated with harmful algal blooms. As a large proportion of the annual nutrient budget of the reservoir is believed to be of riverine origin, nutrient availability is presumably largely influenced by rainfall pattern and hydrological cycles, which are related to changes in climatic conditions such as El Nino-induced drought. The effect of El Nino-induced drought, which occurred in 2015, on the cyanobacterial community structure of Koka Reservoir was, therefore, investigated from May 28, 2015 to April 28 2016. The drought caused the failure of the main rainy season, which expectedly caused reduction in the external input of nutrients and changes in other limnological conditions of the reservoir. These changes, particularly nitrogen limitation, triggered the unusual dominance of the diazotrophic cyanobacterial genus Cylindrospermopsis over the previously persistently dominant nondiazotrophic genus, Microcystis, in the reservoir.

Occurrence of toxic cyanobacteria and cyanotoxins in drinking, recreational, and irrigation waters represents a health hazard to wildlife and humans. This review describes the most common cyanotoxins and their producing taxa and the potential environmental health risks in marine and freshwaters of Saudi Arabia. The most common genera of toxigenic cyanobacteria detected in Saudi waters included Anabaena/Dolichospermum, Anabaenopsis, Cylindrospermopsis, Lyngbya, Microcystis, and Oscillatoria. The cyanotoxins produced by these taxa and have been detected in Saudi Arabia are microcystins (MCYSTs), cylindrospermopsin (CYN), saxitoxins, and lyngbyatoxin-like toxins. Anatoxin has not been investigated in Saudi Arabia, and extensive research is running for this toxin. Studies have detected microcystins in treated drinking waters, groundwater wells, and hot springs and their accumulation in vegetables and fish. Toxin amounts in these commodities often exceeded the WHO guidance values for safe exposure. This indicates that a human health risk could be occurring upon exposure to such potent toxins. Beyond a comprehensive analysis of cyanotoxins in marine and freshwater environments, this paper also identifies and prioritizes research gaps and suggests future research on cyanotoxins in Saudi Arabia.

Toxic cyanobacteria (TCB) are well-known worldwide for their adverse impacts on humans. Species compositions and seasonal variations of TCB in reservoirs depend on interactions between physical and chemical factors. This study was conducted to evaluate the water quality in the Aha Reservoir, Southwest China, focusing on cyanobacteria and cyanotoxins. Water samples were collected weekly or biweekly from May to September of 2015 and used to delineate temporal variations in density and distribution of toxic cyanobacteria and cyanotoxins in the reservoir. Toxic cyanobacteria identified consisted of Aphanizomenon flos-aquae, Pseudanabaena limnetica, Cylindrospermopsis sp., and Microcystis sp., with Aphanizomenon flos-aquae and Pseudanabaena limnetica being the most common and significant toxic genera. The total biomass of cyanobacteria was 17.0 mg/L. Identification and quantification of microcystin variants were conducted by high performance liquid chromatography (HPLC) using a system equipped with a photodiode array detector. Microcystin levels were between 0-3.0 μg/L, MC-RR was around 0-3.0 μg/L and MC-LR was approximately 0-0.9 μg/L. Overall, the results of this study indicate that the investigated reservoirs should be monitored regularly to minimize potential health risks to the human population.

Eutrophication and enhanced external nutrient loading of lakes and seas are most clearly reflected by increased cyanobacterial blooms, which are often toxic. Freshwater cyanobacteria produce a number of bioactive secondary metabolites, some of which have allelopathic properties, significantly influencing the biological processes of other algae, thereby affecting species composition and succession of the phytoplankton. The goal of this work was to investigate the influence of bloom-forming cyanobacterial exudates on the photophysiology of the green alga Scenedesmus quadricauda by chlorophyll fluorescence analysis. We were able to prove the effect of algal cell-free filtrates on the performance of S. quadricauda and demonstrate for the first time that the freshwater picocyanobacterium Cyanobium gracile has strong negative impact on the coexisting green alga. Neither the cyanotoxin (MYC, CYN and ATX) producing, nor the non-toxic strains showed any systematic effect on the production of S. quadricauda. Various strains of the cyanobacterium Cylindrospermopsis raciborskii inhibited the performance of the green alga independently of their origin. Our results urge further studies for a better understanding of the factors affecting the release of allelopathic compounds and the mechanisms of their effects on target organisms.

A wide variety of cyanobacterial species that inhabit freshwater systems are known to produce diverse toxins and off-flavor compounds during the development of environmentally harmful blooms. However, cyanobacterial community development and toxin production potential have not been well studied. In this study, we examined the taxonomic diversity and saxitoxin production potential of cyanobacteria in the water and sediments of a large river, the North Han River in South Korea, by metagenome analysis using next-generation sequencing (NGS) and molecular biological approaches, respectively. NGS revealed that the entire cyanobacterial community in the study area consisted of 39 genera and 47 species. The most abundant genera were Microcystis, Anabaena, Cyanobium, and Synechococcus, which accounted for more than 90% of the entire community. The saxitoxin production potential of the cyanobacterial community was assessed by detecting the sxtA and sxtG genes related to saxitoxin production. Eleven sxtA and 24 sxtG genes were identified through molecular cloning and sequencing. Phylogenic analysis revealed that three sxtA genes that grouped in one phylogenic branch with Scytonema sp. were distinctly separated from the sxtA genes of Anabaena, Aphanizomenon, Lyngbya, and Cylindrospermopsis. Sixteen of the detected sxtG genes were phylogenically similar to those of Anabaena circinalis (Dolichospermum circinale), Aphanizomenon gracile, and Aphanizomenon flos-aquae. Our study demonstrates the utility of the metagenomics approach for characterizing the natural community structure of cyanobacteria containing diverse and even rare species, and the evaluation of saxitoxin-producing potential in the cyanobacterial community.

To investigate the effect of bisphenol A (BPA) on Cylindrospermopsis raciborskii (Cyanobacteria) and Scenedesmus quadricauda (Chlorophyta), we grew the two species at BPA concentrations of 0, 0.1, 1, 2, 5, 10, and 20 mg/L and examined their growth, lipid peroxidation, antioxidant enzyme activity, and chlorophyll a fluorescence. The 96-h EC50 values (effective concentration causing 50% growth inhibition) for BPA in C. raciborskii and S. quadricauda were 9.663 ± 0.047, and 13.233 ± 0.069 mg/L, respectively. A significant reduction in chlorophyll a concentration was found in C. raciborskii and S. quadricauda when BPA concentrations were greater than 1 and 2 mg/L, respectively. Furthermore, F v/F m, ΔF/F m', and qP decreased significantly at 10 mg/L BPA in C. raciborskii but started to decrease at 10 mg/L in S. quadricauda. The changes in chlorophyll fluorescence parameters (α, rETRmax) that were obtained from the rapid light response curves of both algae species showed similar responses to F v/F m, ΔF/F m', and qP under BPA-induced stress. Values for all of the chlorophyll fluorescence parameters in S. quadricauda were higher than in C. raciborskii; however, the nonphotochemical quenching measured in C. raciborskii was considerably higher than it was in S. quadricauda. In addition, lipid peroxidation (determined as MDA content) and antioxidant enzyme activities (SOD and CAT) increased in both species as the BPA concentration increased. These results suggest that C. raciborskii is more sensitive to the effects of BPA than S. quadricauda and that photosystem II might be a target for the activity of BPA in vivo.

It is assumed that daphnids adjust the filter screen morphology in order to minimize the interference with cyanobacterial filaments. The aim of this study was to investigate the impact of filamentous cyanobacteria (Aphanizomenon gracile Lemmermann, Cylindrospermopsis raciborskii Woloszynska Seenaya et Subba Raju) on the thickness and length of setae of the third pair of thoracic limbs of Daphnia magna. The second objective was to assess whether the setae modifications could improve the performance of daphnids in the presence of cyanobacteria. Three clones of Daphnia magna Straus were cultured with: green algae; green algae with filaments of Cylindrospermopsis; and green algae with filaments of Aphanizomenon. The size and age of animals in the first reproduction cycle as well as the number of offspring were recorded. Setae thickness and length were measured in the central part of each endopodite. Additionally, we analyzed how the changes in setae morphology affect the fitness of experimental animals using the intrinsic rate of population increase calculated with the Euler–Lotka equation. The results showed that the thickness and length of setae increased in the presence of filamentous cyanobacteria. Moreover, cyanobacteria-induced setae thickening was positively correlated to the fitness of daphnids, which may indicate setae thickening as a phenotypic adaptation to cope with food stress caused by filamentous cyanobacteria.

Cylindrospermopsis raciborskii and its highly similar relatives Raphidiopsis species have been recognized as globally invasive and expansive filamentous cyanobacteria causing water blooms. Reports on C. raciborskii/Raphidiopsis species and their harmful metabolites such as hepatotoxic cylindrospermopsins (CYNs) in Chinese waters have been increasing, but mostly restricted to the southern regions of China. To further explore the existence and distribution of C. raciborskii in China, six water samples from Beijing city were morphologically and molecularly examined. Five samples of the six were shown to have Cylindrospermopsis filaments with straight and spiral morphotypes. PCR detection targeting on Cylindrospermopsis/Raphidiopsis specific 16S rRNA gene region also showed the positive amplification, and such amplifications were confirmed by sequencing and phylogenetic analysis. As well, three of the five Cylindrospermopsis containing samples were shown to have cyrJ — a gene of CYN synthesis gene cluster. The results represented the presence of toxic Cylindrospermopsis at the most northern line in China so far, indicating rapid expansion of this harmful invasive cyanobacterium. It is strongly suggested that the monitoring on C. raciborskii/Raphidiopsis species and their production of cylindrospermopsin should be emphasized in Beijing and even more northern parts of China.

Rising lake temperatures and changing nutrient inputs are believed to favour the spread of a toxic invasive cyanobacterium, Cylindrospermopsis raciborskii (Woloszynska) Seenayya and Subba Raju, in temperate lakes. However, most evidence for these hypotheses is observational or based on physiological measurements in monocultures. We lack clear experimental evidence relating temperature and nutrients to the competitive success of C. raciborskii. To address this, we performed a 2 × 2 factorial laboratory experiment to study the dynamics of mixed phytoplankton communities subjected to different levels of temperature and phosphorus over 51 days. We allowed C. raciborskii to compete with ten different species from major taxonomic groups (diatoms, green algae, cryptophytes, and cyanobacteria) typical of temperate lakes, under low and high summer temperatures (25 and 30 °C) at two levels of phosphorus supply (1 and 25 µmol L−1). Cylindrospermopsis raciborskii dominated the communities and strongly decreased diversity under low-phosphorus conditions, consistent with the hypothesis that it is a good phosphorus competitor. In contrast, it remained extremely rare in high-phosphorus conditions, where fast-growing green algae dominated. Surprisingly, temperature played a negligible role in influencing community composition, suggesting that changes in summer temperature may not be important in determining C. raciborskii’s spread.

Biological invasions are a major threat to biodiversity and ecosystem functioning. Successful invasions depend on the interplay of multiple abiotic and biotic factors, however, the process of the invasion itself is often overlooked. The temporal variation of environmental factors suggests that a ‘window of opportunity’ for successful invasions exists. Especially aquatic habitats, like temperate lakes, undergo pronounced seasonal fluctuations and show temporally varying environmental conditions in e.g. nutrient availability, temperature and the composition of the resident community including competitors and consumers. We experimentally tested if an invasion window for the globally invasive cyanobacterium Cylindrospermopsis raciborskii exists. From May to September, we determined the invasion success of C. raciborskii in laboratory mesocosms with natural lake water. Although the invasion success was generally low, the invasiveness varied among months and differed in total invasive biomass, net development and final share of C. raciborskii in the community. During the first days, C. raciborskii strongly declined and this initial, short-term decline was independent of the ambient consumptive pressure. These results are in contrast to laboratory studies in which C. raciborskii successfully invaded, suggesting that a complex natural system develops a resistance to invasions.

Cyanobacterial blooms represent a health hazard worldwide due to their toxic potential. The CyanoSurvey project was set up to evaluate the risk associated with the presence of toxigenic cyanobacteria in Australian drinking water reservoirs. It mainly targeted members of the following genera: Dolichospermum, Sphaerospermopsis, Chrysosporum, Cylindrospermopsis and Microcystis. In 2010 and 2011, 64 samples were collected nationally and 196 isolates were purified using several culture media and a concentration technique. The strains obtained were then cryopreserved using three different cryoprotectants. The efficacy of each medium, the applicability of a pre-treatment to the sample and the various cryoprotective agents were compared for several isolates of cyanobacteria. This study highlighted that the ASM-1 medium, used routinely by many laboratories for the isolation and maintenance of cyanobacteria, had the lowest efficacy for the isolation of cyanobacteria from environmental samples. Moreover, the use of a range of media combined with a concentration technique can significantly improve the chances of isolating and purifying certain groups of cyanobacteria. The use of 5 % DMSO for cryopreservation was shown to be a preferable choice of cryopreservative for most strains tested.

This paper presents a case study of a massive fish mortality during a Cylindrospermopsis raciborskii bloom in Aleksandrovac Lake, Serbia in mid-December 2012. According to a preliminary investigation of the samples taken on November 6 before the fish mortalities and to extended analyses of samples taken on November 15, no values of significant physicochemical parameters emerged to explain the cause(s) of the fish mortality. No industrial pollutants were apparent at this location, and results excluded the likelihood of bacterial infections. Even after freezing, the dissolved oxygen concentration in the water was sufficient for fish survival. High concentrations of chlorophyll a and phaeophytin occurred in the lake, and phytoplankton bloom samples were lethal in Artemia salina bioassays. A bloom of the cyanobacterium C. raciborskii was recorded during November. Although the A. salina bioassays indicated the presence of toxic compounds in the cyanobacterial cells, the cyanotoxins, microcystins, cylindrospermopsin and saxitoxin were not detected.

Members of the genus Cylindrospermopsis represent an important environmental and health concern. Strains CS-508 and MVCC14 of C. raciborskii were isolated from freshwater reservoirs located in Australia and Uruguay, respectively. While CS-508 has been reported as non-toxic, MVCC14 is a saxitoxin (STX) producer. We annotated the draft genomes of these C. raciborskii strains using the assembly of reads obtained from Illumina MiSeq sequencing. The final assemblies resulted in genome sizes close to 3.6 Mbp for both strains and included 3202 ORFs for CS-508 (in 163 contigs) and 3560 ORFs for MVCC14 (in 99 contigs). Finally, both the average nucleotide identity (ANI) and the similarity of gene content indicate that these two genomes should be considered as strains of the C. raciborskii species. Electronic supplementary material The online version of this article (10.1186/s40793-018-0323-1) contains supplementary material, which is available to authorized users.

Cylindrospermopsis raciborskii is a toxic cyanobacterium with an invasive nature. The species is found in all the main continents but its origin and dispersal routes on a worldwide perspective remain yet mostly unknown. In this study, 27 isolates of C. raciborskii gathered worldwide have been used for an in-deep phylogenetic analyses with a concatenated system of three genetic markers (16 rRNA, 16S-23S ITS larger subunit, and RNA polymerase rpoC1) comprehending 3,188 bp. Our results provide support for an origin of C. raciborskii in the American continent. Dispersal routes included afterward a spread into the African continent and then Asia and Australia, being Europe the last continent to be colonized by this species. Our phylogenetic inferences suggest that C. raciborskii seem to have a well-defined dispersal behavior with a well-established population structure around the world.

The tropical cyanobacterium Cylindrospermopsis raciborskii is of particular concern for its invasive characteristics and production of the toxin cylindrospermopsin (CYN). The present study represents the first attempt to determine the distribution of C. raciborskii and CYN in tropical China. The presence of C. raciborskii and CYN, as well as the composition of phytoplankton, was determined from a total of 86 samples from 25 urban reservoirs for drinking water supply in Dongguan City of South China. The presence of C. raciborskii was observed in 21 of the 25 reservoirs and confirmed that this species has been widely distributed in the investigated reservoirs. C. raciborskii accounted for between 0.1 and 90.3 % of the total phytoplankton biomass and contributed to the majority of the phytoplankton in some reservoirs such as Tangkengbian and Xiagongyan. Its biomass was negatively correlated with NO3 −-N concentration and Secchi depth. Dissolved CYN was detected in more than one-half of the reservoirs with concentrations up to 8.25 μg L−1, and it positively correlated with C. raciborskii biomass. Dissolved microcystins (MCs) were detected in 12 of the 25 reservoirs with a maximum concentration 1.99 μg L−1. Our data strongly suggest that C. raciborskii and CYN could be important health hazards in urban reservoirs of South China and that more data are needed for further assessment.

The Macau Storage Reservoir (MSR) has experienced algal blooms in recent years, with high levels of Cylindrospermopsis and Microcystis and detectable concentrations of cyanotoxins. To analyze the cyanotoxin-producing genotypes and relate the corresponding cyanotoxins to the water quality parameters, a quantitative real-time polymerase chain reaction (qPCR) was developed and applied to the water samples in three locations of MSR. Cylindrospermopsin synthetase gene (pks) and a series of microcystin synthetase genes (mcys) were used for identifying and quantifying cylindrospermopsin- and microcystin-producing genes, and the corresponding water parameters were measured accordingly.

Monitoring of cyanobacteria and their toxins are traditionally conducted by cell counting, chlorophyll-a (chl-a) determination and cyanotoxin measurements, respectively. These methods are tedious, costly, time consuming, and insensitive to rapid changes in water quality and cyanobacterial abundance. We have applied and tested an online phycocyanin (PC) fluorescence probe for rapid monitoring of cyanobacteria in the Macau Storage Reservoir (MSR) that is experiencing cyanobacterial blooms. The relationships among cyanobacterial abundance, biovolume, cylindrospermopsin concentration, and PC fluorescence were analyzed using both laboratory and in-the-field studies. The performance of the probe was compared with traditional methods, and its advantages and limitations were assessed in pure and mixed cyanobacterial cultures in the laboratory. The proposed techniques successfully estimated the species including Microcystis and Cylindrospermopsis, two toxic species recently observed in the MSR. During February–November, 2010, the PC probe detected high correlations between PC and cell numbers (R 2 = 0.71). Unlike the chl-a content, which indicates only the total algal biomass, the PC pigment specifically indicates cyanobacteria. These results support the PC parameter as a reliable estimate of cyanobacterial cell number, especially in freshwater bodies where the phytoplankton community and structure are stable. Thus, the PC probe is potentially applicable to online monitoring of cyanobacteria.

The toxic cyanobacterial communities of four recreational reservoirs (Bueng Kaen Nakhon, Bueng Thung Sang, Bueng Nong Khot, and Bueng See Than) in Amphur Muang, Khon Kaen Province, Thailand, were investigated. Water samples were collected via monthly sampling from June to October 2011 for the study on the diversity and density of toxic cyanobacteria and toxin quantification. The main toxic cyanobacteria present in these reservoirs were Aphanocapsa sp., Cylindrospermopsis sp., Leptolyngbya sp., Limnothrix sp., Microcystis sp., Oscillatoria sp., Planktolyngbya sp., Planktotrix sp., and Pseudanabaena sp. The dominant bloom-forming genera in the water samples from Bueng Nong Khot and Bueng See Than were Microcystis sp. and Cylindrospermopsis sp., respectively. Enzyme-linked immunosorbent assays specific for cyanotoxins were performed to detect and quantify microcystins and cylindrospermopsins, with the highest average microcystins content (0.913 μgL(-1)) being found in the sample collected from Bueng Nong Khot and the highest average cylindrospermopsins content (0.463 μgL(-1)) being found in the sample collected from Bueng See Than. The application of 16S rRNA analyses to cyanobacterial isolates BKN2, BNK1, BNK2, and BST1 indicated that these isolates are most closely related to Limnothrix planctonica (JQ004026) (98% similarity), Leptolyngbya sp. (FM177494) (99% similarity), Microcystis aeruginosa (DQ887510) (99% similarity), and Limnothrix redekei (FM177493) (99% similarity), respectively.

In order to investigate the effect of the allelopathic compound pyrogallic acid on Cylindrospermopsis raciborskii F2, the impact on growth, oxidative stress and expression of the psbA, grpE, fabZ, recA, cmpA, ftsZ and cyrJ genes were studied. The results indicated significant decreases in Chl a and cell number following a 24-h incubation with 4 mg L(-1) pyrogallic acid. Additionally, malodialdehyde content, superoxide dismutase activity and catalase activity were enhanced following treatment with 2 and 4 mg L(-1) pyrogallic acid. Expressions of the genes psbA, grpE, fabZ, recA and cyrJ were significantly up-regulated following exposure to 4 mg L(-1) pyrogallic acid, while no changes were observed with concentrations of 1 or 2 mg L(-1). Expression of cmpA was significantly down-regulated following treatment with the lowest tested concentration of pyrogallic acid (1 mg L(-1)), while ftsZ was only significantly down-regulated with concentrations of 2 and 4 mg L(-1). These results suggest that photosynthesis inhibition and oxidative damage are important modes of action for the allelopathic effect of pyrogallic acid on C. raciborskii.

The responses of the snail central neurons (Helix pomatia, Lymnaea stagnalis) and the isolated Helix heart were characterized evoked by cyanobacterial extracts (Cylindrospermopsis raciborskii ACT strains) isolated from Lake Balaton (Hungary). The nicotinergic acetylcholine (ACh) receptors in the CNS (both excitatory and inhibitory) were blocked by the extracts of ACT 9502 and ACT 9505 strains and the anatoxin- a (homoanatoxin-a) producing reference strain of Oscillatoria sp. (PCC 6506), similar to the inhibitory effects of the pure anatoxin-a. The enhancement of the ACh responses by the ACT 9504 extract suggests additional, probably acetylcholine esterase inhibitory mechanisms. On the isolated Helix heart the crude ACT 9505 and PCC 6506 extracts evoked frequency increase and transient twitch contraction, opposite to the ACh evoked heart relaxation. Anatoxin-a similarly contracted the heart but did not increase its contration frequency. These data suggest the involvement of some non-cholinergic mechanisms, acting very likely by direct modulation of the electrical or contractile system of the isolated heart. Diversity of the effects evoked by the cyanobacterial extracts in the CNS and heart suggest pharmacologically different neuroactive components among the secondary metabolites of the cyanobacteria acting on both (anatoxin-a like) cholinergic and (unidentified) non-cholinergic receptors.

This study reports the presence of the cyanobacterial toxin cylindrospermopisn (CYN) and its producer Cylindrospermopsis raciborskii for the first time in Saudi freshwater sources. C. raciborskii was found in Gazan Dam Lake water with two morphotypes (coiled and straight). The appearance and cell density of this species was significantly positively related to high temperature and high ammonium concentrations, and negatively with nitrate and phosphate concentrations in the lake. Intracellular concentrations of CYN (4-173 μg L(-1)) were associated with C. raciborskii rather than other cyanobacteria with a maximal value obtained in June 2011, coinciding with the highest bloom of this species (19 × 10(7) trichome L(-1)). CYN cell quotas (0.6-14.6 pg cell(-1)) varied significantly along the study period and correlated with most environmental factors. The results of ELISA and liquid chromatography-mass spectrometry proved that the CYN production by strains of this species was isolated from this lake during the present study, with an amount reaching 568 μg g(-1). Extracellular CYN was also detected in cell-free lake water at concentrations 0.03-23.3 μg L(-1), exceeding the drinking water guideline value of 1 μg L(-1) during the Apr-Jul period. As this lake is an important source for drinking and irrigation waters, CYN monitoring should be included in the environmental and health risk assessment plans of these water bodies.

The increasing contamination of aquatic environments motivates studies on the interactions among natural dissolved organic matter, metals, and the biota. This investigation focused on the organic exudates of the toxic cyanobacteria Cylindrospermopsis raciborskii as a Cu carrier through a three-level aquatic trophic chain (bacteria, protozoa, and copepod).The effects of bacteria activity and growth on the metal-organic complexes were evaluated through changes in free Cu(2+) ions, total dissolved, and total particulate Cu. To be sure that the added copper would be complexed to the exudates, its complexing properties were previously determined. The cyanobacteria exudate-Cu complexes were furnished to bacteria that were further used as a food source to the protozoan Paramercium caudatum. This was then furnished as food to the copepod Mesocyclops sp. The results showed that, in general, the cyanobacterial exudates decreased Cu bioavailability and toxicity to the first trophic level (bacteria), but because the heterotrophic bacteria accumulated Cu, they were responsible for the transference for the otherwise low availability metal form. Both the bacteria and protozoan organisms accumulated Cu, but no metal accumulation was detected in the copepods.

A member of the cyanobacterial genus Cylindrospermopsis Seenayya et Subba Raju (Order Nostocales) invaded Lake Kinneret (Israel) in 1998. Since then it has been found in the water column nearly every summer, initially at low concentrations, but since 2003 it formed major summer blooms. The goals of the present study were to identify the invading species based on its morphology, annual life cycle, toxicity, genetic markers, and its phylogeny, and to examine its range of morphological variability. Cell counts, allometric measurements, and filament morphology were determined on samples collected weekly from a mid-lake station throughout 2005. Akinetes, heterocytes, filament end types, cellular contents, and the annual cycle of filament morphology were described, supporting the identification as Cylindrospermopsis raciborskii (Woloszynska) Seenayya et Subba Raju. Cells had many small, few large, or no dark granules presumed to be polyphosphate bodies. Filament fragmentation was observed and six types of filament ends were described. Sequence analysis of the 16SrRNA and ITS-L DNA fragment from a laboratory isolate of the cyanobacterium also confirmed the identification as C. raciborskii. Biomass of this species collected from Lake Kinneret during a bloom did not show toxicity in Artemia bioassays, and the toxin cylindrospermopsin was not detected in a methanolic extract of the isolate. Genetic examination indicated that C. raciborskii from Lake Kinneret lacked the cyrJ component of the cylindrospermopsin synthase cluster and, thereby, was incapable of producing the toxin. Due to the morphological plasticity of this species, the identification of C. raciborskii from Lake Kinneret was not straight forward and required taking multiple approaches, including microscopic observations over a full annual cycle, culture isolation, and molecular taxonomy.

Static-renewal ecotoxicity trials monitored growth of Hydrilla verticillata in conjunction with exposure to Cylindrospermopsis raciborskii whole-cell extracts containing the cyanotoxin cylindrospermopsin (CYN). Maximum exposure concentrations were 400 microg L(-1) CYN over 14 days. The responses of Hydrilla to the treatments were variable according to the toxin concentrations and lengths of exposure. Plant deaths, chlorosis, and necrosis were not recorded from treated plants. However, Hydrilla experienced significant growth stimulation and redistribution of plant resources in conjunction with exposure to the whole-cell extracts. Root production was particularly impacted. The results of this study imply that root production could aid in reducing C. raciborskii cell concentrations and CYN toxicity. Results of chlorophyll analyses differed, indicating that CYN in whole-cell extracts might exert complex effects on photosynthesis. This is the first study to describe the responses of an aquatic macrophyte following exposure to C. raciborskii whole-cell extracts containing the cyanotoxin cylindrospermopsin.

Cylindrospermopsis raciborskii, an invasive freshwater cyanobacterium, originated from the tropics but has spread to temperate zones over the last few decades. Its northernmost populations in Europe occur in North German lakes. How such dramatic changes in its biogeography are possible and how its population dynamics in the newly invaded habitats are regulated are still unexplained. We therefore conducted a long-term (1993-2005) study of two German lakes to elucidate the mechanisms behind C. raciborskii population dynamics and to identify the abiotic constraints on its development. Our data revealed that pelagic populations of C. raciborskii thrived for three months during the summer, contributing up to 23% of the total cyanobacteria biovolume. Population sizes varied greatly between years without exhibiting any distinct long-term trends. In the annual lifecycle, C. raciborskii filaments emerged in the pelagic habitat when the temperature rose above 15-17 degrees C. At that time, mean photosynthetically active radiation in the mixed water column (I (mix)) overstepped its maximum. Rates of population net increase were highest at the beginning of the season (0.15-0.28 day(-1)), declined continuously over time, and were significantly positively correlated with I (mix). This indicates that the onset of the pelagic population is temperature-mediated and that I (mix) controls its growth. Since I (mix) peaks before the population onset, the time of germination is of crucial importance for successful development. To test this hypothesis, we designed a model to simulate pelagic population size, starting at different dates in the annual cycle. Moving the population onset forward by 30 days resulted in a doubling of the population size. We therefore conclude that an earlier rise in water temperature associated with climate change has promoted the spread of C. raciborskii to the temperate zone. Earlier warming permits earlier germination, thereby shifting the pelagic populations to a phase with higher I (mix), which advances growth and the population establishment.

Zooplankton can influence the phytoplankton community through preferential grazing. In turn, nuisance cyanobacteria may affect zooplankton community structure by allowing certain species to out-compete others. We examined zooplankton-phytoplankton interactions, micro-zooplankton (< 200 μm) grazing, and biochemical components of the seston in the St. Johns River System (SJR), Florida in the presence and absence of the toxin-producing cyanobacterium Cylindrospermopsis raciborskii. We tested whether this cyanobacterium would cause a decrease in the size structure of the zooplankton community and postulated a resultant decline in the metabolic energy and carbon available to higher consumers (i.e. fish). When numbers of C. raciborskii were low or undetectable, zooplankton were more diverse and were comprised of larger species. Rotifers were the dominant zooplankton, and their numbers relative to other zooplankton increased as C. raciborskii concentrations increased. Micro-zooplankton grazing was higher in times of C. raciborskii abundance, suggesting competitive and predatory exclusion by larger zooplankton in times of higher phytoplankton diversity. Total caloric content of the seston was higher in times of C. raciborskii abundance. However, essential fatty acids and phosphorus may be lacking in the seston, or nutrients may potentially be sequestered by the cyanobacteria and remain as organic matter in the water column. In such cases, higher trophic levels would not be able to obtain optimal energy requirements. Overall, there was a greater impact of micro-grazers on phytoplankton in the presence of C. raciborskii and apparent negative effects on the larger zooplankton species, suggesting a potential for changes in zooplankton and higher trophic level community structure.

Salinity has been suggested as being a controlling factor for blooms of N 2 -fixing cyanobacteria in estuaries. We tested the effect of salinity on the growth, N 2 fixation, and photosynthetic activities of estuarine and freshwater isolates of heterocystous bloom-forming cyanobacteria. Anabaena aphanizomenoides and Anabaenopsis sp. were isolated from the Neuse River Estuary, North Carolina, and Cylindrospermopsis raciborskii from Lakes Dora and Griffin, central Florida. Salinity tolerance of these cyanobacteria was compared with that of two Nodularia strains from the Baltic Sea. We measured growth rates, N 2 fixation (nitrogenase activity), and CO 2 fixation at salinities between 0 and 20 g L -1 NaCl. We also examined photosynthesis-irradiance relationships in response to salinity. Anabaenopsis maintained similar growth rates in the full range of salinities from 2 to 20 g L -1 NaCl. Anabaena grew at up to 15 g L -1 , but the maximum salinity 20 g L -1 NaCl was inhibitory. The upper limit for salinity tolerance of Cylindrospermopsis was 4 g L -1 NaCl. Nodularia spp. maintained similar growth rates in the full range of salinities from 0 to 20 g L -1 . Between 0 and 10 g L -1 , the growth rate of Nodularia spumigena was slower than that of the Neuse Estuary strains. In most strains, the sensitivity of nitrogenase activity and CO 2 fixation to salinity appeared similar. Anabaenopsis, Anabaena, and the two Nodularia strains rapidly responded to NaCl by increasing their maximum photosynthetic rates (P max ), Overall, both Neuse River Estuary and Baltic Sea strains showed an ability to acclimate to salt stress over short-(24 h) and long-term (several days to weeks) exposures. The study suggested that direct effect of salinity (as NaCl in these experiments) on cyanobacterial physiology does not alone explain the low frequency and magnitude of blooms of N 2 -fixing cyanobacteria in estuaries.

Cyanobacterial dominance in lakes has received much attention in the past because of frequent bloom formation in lakes of higher trophic levels. In this paper, underlying mechanisms of cyanobacterial dominance are analyzed and discussed using both original and literature data from various shallow mixed and deep stratifying lakes from temperate and (sub)tropical regions. Examples include all four ecotypes of cyanobacteria sensu Mur et al. (1993), because their behavior in the water column is entirely different. Colony forming species (Microcystis) are exemplified from the large shallow Tai Hu, China. Data from a shallow urban lake, Alte Donau in Austria are used to characterize well mixed species (Cylindrospermopsis), while stratifying species (Planktothrix) are analyzed from the deep alpine lake Mondsee. Nitrogen fixing species (Aphanizomenon) are typified from a shallow river-run lake in Germany. Factors causing the dominance of one or the other group are often difficult to reveal because several interacting factors are usually involved which are not necessarily the same in different environments. Strategies for restoration, therefore, depend on both the cyanobacterial species involved and the specific causing situation. Some uncertainty about the success of correctives, however, will remain due to the stochastic nature of the events and pathways leading to cyanobacterial blooms. Truly integrated research programs are required to generate predictive models capable of quantifying key variables at appropriate spatial and temporal scales.